Directeur de la rédaction Pr François Paille Rédacteur en chef Pr Amine Benyamina Rédacteurs associés Dr Philippe Batel Dr Ivan Berlin Dr Laurent Karila Pr Michel Lejoyeux Pr Mickaël Naassila Rédactrice Sciences humaines Pr Myriam Tsikounas Rédactrice Sciences psychologiques Pr Isabelle Varescon-Pousson Comité de rédaction Pr Georges Brousse Pr Olivier Cottencin Dr Michel Craplet Pr Jean-Bernard Daeppen Dr Jean-Michel Delile Pr Maurice Dematteis Dr Claudine Gillet th Dr Geneviève Lafaye 17 European Society for Biomedical Research Pr Michel Reynaud Dr Alain Rigaud Dr Marc Valleur on Congress Directeur de la publication Pr Mickael Naassila 21-24 September 2019, Lille Comité scientifique Pr Jean Adès Pr Thomas F. Babor Pr Jean-Louis Balmès Pr Maurice Bazot Dr Mats Berglund Pr Jacques Besson Pr Jean-Pierre Blayac Pr Jonathan D. Chick Mme Marie Choquet Pr Philippe de Witte Pr Michel Escande Pr Claude Got Dr Antoni Gual Pr Momar Gueye Pr Roger Henrion Pr Denise Kandel Pr Michel Le Moal Pr Karl Mann Mme Véronique Nahoum-Grappe Dr José Maria Neves Cardoso Pr Philippe-Jean Parquet Pr Jean-Louis Pedinielli Pr Falvio Poldrugo Pr Bernard Roques Pr John A. Talbott Pr Jean-Luc Vénisse Pr Lars von Knorring Pr Jacques Weill Pr Jean-Jacques Yvorel

ISSN 2554-4853 Trimestriel Société Française

PRINCEPS Éditions SEPTEMBRE-DÉCEMBRE 2019 - Tome 41, n° 3-4 d’Alcoologie               

           !BSTRACTS CONGRÈS

Pr Mickael Naassila* * President 17th ESBRA Meeting

17th European Society for Biomedical Research on Alcoholism congress 21-24 September 2019, Lille – Invited talks and Symposia abstracts

Invited talks forward, such as combining training with neurostimulation. Together these fndings emphasize the malleability of the addicted brain and the promise of targeted CT in the treat- Targeting biased decision making in the treatment ment of AUD. of use disorders Reinout Wiers (Amsterdam, The Netherlands) ASH-NASH synergism and its underlying mechanisms Alcohol Use Disorder (AUD) and other have been Hidekazu Tsukamoto (Los Angeles, USA) characterized as a chronic brain disease from the biomedical perspective and as the unfortunate outcome of adverse social Alcohol misuse and obesity are two leading independent risk conditions from the social science perspective. We emphasize factors for alcoholic and non-alcoholic steatohepatitis (ASH biased decision making as a central characteristic in (alcohol) and NASH) around the globe. Synergistic interactions by . From a therapeutic perspective, the important these factors have also increasingly been recognized. In fact, question is to what extent these biases reverse after successful the emerging evidence indicates the average BMI of some abstinence, and to what extent they can be reversed through ASH patient populations in the US may be around 30. ASH targeted training. Two types of Cognitive Training (CT) and alcoholic cirrhosis occur as the consequences of alco- can be distinguished: those in which general abilities are hol addiction which dictates heavy drinking and sustained trained (e.g., working memory training) and those in which blood alcohol levels (BAL) due to physical dependence. Tis initial motivational reactions to alcohol are targeted, so called condition can be reproduced in rodents by intragastric fee- cognitive biases (Cognitive Bias Modifcation, CBM, Wiers ding of ethanol diet which also allows precise reproduction 2018). I will review the state of afairs in both. Training of of the synergism between sustained BAL and overfeeding- general abilities takes a long time, but does show promise for induced obesity. Tis model exhibits heightened steatohepa- a subgroup of patients. CBM has shown to increase one-year titis, M1 macrophage activation, nitrosative stress driven by abstinence in several large clinical trials, with efect sizes Notch-dependent mitochondrial metabolic reprogramming. similar to medication for alcohol (NNT=12). It is also beco- Moderate alcohol intake may also synergistically work with ming clear for which individuals CBM shows most promise NASH to promote liver cancer development. Evidence sug- as an add-on treatment (those with a strong cue-reactivity gests social drinking is sufcient enough to promote live can- and/or impulsivity), and we are beginning to understand cer incidence in NASH-cirrhosis patients. Tis synergism is the neurocognitive mechanisms underlying training efects reproduced in mice injected with the hepatocarcinogen DEN (e.g., reduced cue-reactivity). CT shows modest but reliable and fed alcohol-containing Western diet. Tumor promotion efects as add-on to regular psychosocial treatment, but does in this model is dependent on activation of hepatic stellate not appear to work in the absence of psychosocial treatment, cells (HSC) driven by Wnt--catenin-mediated overexpres- nor in the absence of motivation to change (e.g. in proof- sion of stearoyl-CoA-desaturase (SCD), which in turn esta- of-principle studies in students). Finally, I will sketch ways blishes a SCD-LRP5/6-Wnt positive loop to amplify Wnt-

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-catenin pathway in HSC and tumor microenvironment, etheno DNA-adducts and the severity of fbrosis. First results leading to tumor-promoting lipid metabolic reprogramming. of the efect of CYP2E1 inhibition by chlormethiazole, a Tese fndings highlight the causal roles of morphogen- specifc CYP2E1 inhibitor on ALD can be expected soon. driven metabolic reprogramming in steatohepatitis and liver tumor development promoted by ASH-NASH synergism. Young investigator symposium Applying precision medicine to alcohol and use disorders Henry R Kranzler1 (1 Philadelphia, USA) increases alcohol self-administration via µ- receptor activity in the ventral tegmental Although most medications approved to treat alcohol use area disorder (AUD) and (OUD) have been E. Domi1, A. Hansson2, P. Marvin2, E.Barbier1, Xu Li1, E. Augier1, shown to be efcacious in placebo-controlled trials, efect M. Heilig1 (1 Linkoping, Sweden, 2 Mannheim, Germany) sizes vary among them. Even medications with the largest efect sizes, however, are not efcacious for all (or perhaps Alcohol and nicotine are the most commonly co-abused even most) patients treated with them. Recent eforts to en- , with a large majority of alcoholics diagnosed with a hance the therapeutic efects of these medications have used comorbid addiction to nicotine. Te endogenous opioid sys- a precision medicine approach, including the use of pharma- tem is involved in the rewarding properties of both alcohol cogenetics (PGx) to identify genetic predictors of treatment and nicotine. We previously found that CERC-501, a highly response. Tis lecture will discuss recent developments in the selective KOR antagonist reduced escalated alcohol self-ad- PGx of AUD and OUD. Specifc topics to be covered are: ministration induced by the intermittent access to alcohol 1) a variant in the mu- gene (OPRM1) and 20%. In here we tested the efect of CERC-501 on escalation response to treatment of AUD, 2) a variant in the of alcohol drinking induced by nicotine. Chronic nicotine eli- kainate receptor gene and response to topiramate treatment cited a robust and specifc escalation of alcohol drinking wit- of AUD, 3) a variant near OPRM1 and usual hout afecting saccharin self-administration and locomotion. dose for treating OUD, and 4) a variant in the delta-opioid CERC-501 did not suppress nicotine-induced increased alco- receptor gene (OPRD1) and response to hol self-administration in opposite to naltrexone which bloc- treatment of OUD. Findings from these studies underscore ked escalated drinking. Our in situ hybridization data showed the potential utility of a precision medicine approach to trea- a diferent pattern of expression and functional activation of ting alcohol and drug use disorders and some of the obstacles MORs in alcohol escalation induced by nicotine, while KORs to be overcome in advancing the feld. expression and activity was not afected by the combination The role of CYP2E1 in alcoholic liver disease and of the two drugs. Specifcally, our data showed an increased expression and a decreased function of MORs in the ventral alcohol mediated carcinogenesis 1 1 1 tegmental area of alcohol-escalated rats. Nicotine induced Helmut K. Seitz , Sebastian Mueller ( Heidelberg, Germany) escalation of alcohol self-administration was also accompa- nied by decreased p-DARPP32 in nucleus accumbens shell. Various factors are involved in the pathogenesis of alcoholic Tis suggest that nicotine pretreatment reduces the rewarding liver disease (ALD) and ethanol mediated carcinogenesis. In value of alcohol and therefore mediates alcohol escalation. addition to genetic, epigenetic and immunologic mechanisms, In conclusion our results also suggest that targeting µ rather acetaldehyde associated toxicity, oxidative stress as well as than −opioid receptors may represent a promising phar- mediated infammation are of major importance. macotherapeutic approach for the treatment of alcohol use Oxidative stress with the generation of reactive oxygen spe- disorders where alcohol consumption is driven by nicotine. cies (ROS) develops either in infammation (alcoholic hepa- titis) or during oxidation of ethanol via cytochrome P4502E1 Unveiling the alcohol-dependent alterations of (CYP2E1). CYP2E1 is induced by ethanol, oxidizes ethanol local translation in the prefrontal cortex during to acetaldehyde and generates ROS during this process. ROS adolescence results in protein damage, enhanced fbrogenesis and DNA S. Laguesse, L. Van Hees, L. Nguyen (Liege, Belgium) lesions. Furthermore, CYP2E1 induction results in an en- hanced activation of various procarcinogens and an increased Alcohol use disorder (AUD) is a devastating relapsing disease degradation of retinol and retinoic acid (RA), a compound which represents the fourth leading cause of preventable responsible for cell diferentiation and proliferation. An inhi- death worldwide. AUD has mainly been considered as a bition of CYP2E1 results in an improvement of ALD and pathological condition in adults, but recent evidence suggests chemically induced carcinogenesis in animal experiments. In that the roots of alcohol addiction begin to grow during man, CYP2E1 is induced following the consumption of 40 adolescence. Adolescence is a critical developmental period grams of ethanol per day already after one week. However, characterized by signifcant changes in brain architecture the induction varies interindividually. Te mechanism for this and behaviors. Brain maturation begins in posterior regions is still unclear. Patients with ALD show a signifcant correla- and progresses towards anterior higher-order regions, inclu- tion between CYP2E1, the occurrence of highly carcinogenic ding the prefrontal cortex (PFC). Te PFC is implicated

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 233 Congress in executive functions and its immaturity in adolescents alcohol-use disorders but also explain relapse risk (Rupp et is associated with lack of inhibitory control over behavior, al., 2017). Regarding the binge drinking alcohol consump- increased impulsivity and desire of risk-taking. It is widely tion pattern (i.e. alternation between high intoxications and believed that the enhanced ability of the adolescent PFC to withdrawals), research proposed that it would induce similar undergo experience-dependent changes is associated with brain alterations than severe alcohol-use disorders (Stephens heightened vulnerability to exogenous agents, including and Duka, 2008). Tis proposal supports the continuum alcohol. Adolescent Alcohol Exposure (AAE) may interfere hypothesis, suggesting that binge drinkers would be characte- with the ongoing maturation of frontal brain circuits, leading rized by qualitatively similar impairments than patients with to profound long-lasting consequences on PFC structure and severe alcohol-use disorders (Enoch, 2006; Sanhueza et al., function. In addition, AAE is related to serious psychological 2011). Terefore, the existence of emotional impairments in problems, comorbid psychopathology and detrimental neuro- binge drinking could precipitate the development of chronic cognitive consequences, and clinical studies have shown that and severe alcohol-related disorders (e.g., Wills et al., 2016). AAE signifcantly increases the risk of developing psychiatric Accordingly, it appears central that future binge drinking and behavioral disorders later in life, including addiction. studies explore emotional processing and their brain corre- However, the precise cellular mechanisms underlying the lates to precisely determine their role in the maintenance of alcohol-induced cognitive and behavioral impairments, the excessive alcohol use and the possible development of severe molecular mechanisms underlying defects in PFC matura- alcohol-use disorders. Te current literature mainly showed tion, and possible sex diferences are still poorly understood. that when binge drinkers had to process emotional contents Alcohol addiction is considered as a maladaptive form of (e.g., identifcation, recognition), they performed poorly than learning and memory. Indeed, alcohol is thought to “usurp” control participants (Lannoy et al., 2018b; Maurage et al., the molecular mechanisms underlying those processes, 2013a). Disrupted brain activations and electrophysiological including synaptic plasticity, which depends on the local activities were also observed during the identifcation of emo- translation of mRNAs at synaptic sites. It has been shown tional prosodies (Maurage et al., 2009, 2013a) and emotional in adult mice that excessive alcohol consumption modifes crossmodal integration between facial and vocal expressions synaptic protein composition in brain regions associated with (Lannoy et al., 2018a). Moreover, altered electrophysiological the mesocorticolimbic pathway, promoting the development activities were also found during the view of positive and and maintenance of addiction. Here we use a mouse model negative afective scenes (Connell et al., 2015; Huang et al., of voluntary adolescent binge drinking to study the alcohol- 2017). Nevertheless, it is unclear whether the simple view dependent structural and functional defects in the PFC as of emotions (i.e. when no specifc processing is required on well as the behavioral consequences. We report that excessive emotional stimuli) already lead to alterations at the brain alcohol consumption during adolescence leads to long-las- level, as preliminary observed for the processing of afective ting behavioral impairments in adulthood, such as increased scenes. In the current study, we explored, beyond the ability anxiety and alcohol intake as well as reduced cognitive per- to explicitly recognize emotions, the brain activations related formances, both in males and females. By using transgenic to the simple view of emotional stimuli in binge drinkers and mouse lines and Ribotag profling, we are comparing the controls. Behavioral and neuroimaging fndings were combi- synaptic translatome of specifc neuronal populations in the ned to explore brain responses during the view of emotional PFC (i.e. glutamatergic neurons and interneurons) in order to facial expressions (happiness, anger, sadness, fear, contempt) identify candidate synaptic mRNAs whose translation levels while participants performed a gender categorization task. are modifed by AAE. Preliminary analyses showed specifc patterns of activations Neural correlates of implicit emotional processing in binge drinking, such as increased responses in the anterior cingulate cortex during the implicit processing of fear. By in binge drinking highlighting disrupted brain activations whereas no direct Séverine Lannoy1, Fabien Gierski1,2, Laurence Dricot3, Farid 1 1 1 emotions processing is required, these results extend the un- Benzerouk , Christophe Portefaix , Sarah Barrière , Véronique derstanding of emotional difculties in binge drinking. Tey 2 1 2 1 Quaglino , Arthur Kaladjian , Mickael Naassila ( Reims, also support the continuum hypothesis regarding emotional 2 3 , Amiens, France, Brussels, Belgium) alterations between binge drinking and alcohol-use disorders and reinforce that emotional impairments may be considered Binge drinking is a widespread alcohol consumption pattern a central vulnerability factor in alcohol-related disorders. in young people, defned by occasional but high alcohol References intoxications (Courtney and Polich, 2009). It has been related - Carbia, C., López-Caneda, E., Corral, M., Cadaveira, F., 2018. A systema- to deleterious consequences such as brain modifcations and tic review of neuropsychological studies involving young binge drinkers. cognitive dysfunctions (Carbia et al., 2018; Hermens et al., Neurosci. Biobehav. Rev. 90, 332–349. https://doi.org/10.1016/j.neu- 2013; Maurage et al., 2013b). Recent studies have also under- biorev.2018.04.013. - Connell, A.M., Patton, E., McKillop, H., 2015. Binge drinking, depression, lined a difculty to process emotions in young binge drinkers and electrocortical responses to emotional images. Psychol. Addict. Behav. (e.g., Huang et al., 2017; Lannoy et al., 2018b), suggesting 29, 673–682. https://doi.org/10.1037/adb0000071. that this alcohol consumption pattern may also be associated - Courtney, K.E., Polich, J., 2009. Binge drinking in young adults: Data, with emotional impairments. Importantly, emotional impair- definitions, and determinants. Psychol. Bull. 135, 142–156. https://doi. ments have been identifed as key factors to describe severe org/10.1037/a0014414.

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- Enoch, M.-A., 2006. Genetic and environmental influences on the deve- lopment of alcoholism: Resilience vs. risk. Ann. N. Y. Acad. Sci. 1094, we investigated the association between and acylated 193–201. https://doi.org/10.1196/annals.1376.019. ghrelin and mesolimbic brain response to alcohol cues, alco- - Hermens, D.F., Lagopoulos, J., Tobias-Webb, J., De Regt, T., Dore, G., hol craving and relapse risk in a sample of seventy alcohol- Juckes, L., Latt, N., Hickie, I.B., 2013. Pathways to alcohol-induced brain impairment in young people: A review. Cortex. 49, 3–17. https://doi. dependent patients in the post-acute withdrawal phase. org/10.1016/j.cortex.2012.05.021. Methods: a total of seventy abstinent alcohol-dependent - Huang, S., Holcomb, L.A., Cruz, S.M., Marinkovic, K., 2017. Altered oscilla- patients were recruited from an in-patient setting in the tory brain dynamics of emotional processing in young binge drinkers. Cogn. Affect. Behav. Neurosci. https://doi.org/10.3758/s13415-017-0551-7. Clinic of Addictive Behavior and Addiction Medicine of the - Lannoy, S., D’Hondt, F., Dormal, V., Blanco, M., Brion, M., Billieux, J., Central Institute of Mental Health (Mannheim, Germany) Campanella, S., Maurage, P., 2018a. Electrophysiological correlates of after having completed detoxifcation treatment (mean abs- emotional crossmodal processing in binge drinking. Cogn. Affect. Behav. Neurosci. 18, 1076–1088. https://doi.org/10.3758/s13415-018-0623-3. tinence=11.6 days, range=5-25). All patients underwent a - Lannoy, S., Dormal, V., Brion, M., Gaudelus, B., Billieux, J., Maurage, P., combined psychometric and functional magnetic resonance 2018b. Affective impairments in binge drinking: Investigation through imaging (fMRI) assessment of alcohol cue-reactivity and emotional facial expression decoding. Compr. Psychiatry. 83, 59–63. https:// alcohol craving using a validated set-up (Vollstadt-Klein ., doi.org/10.1016/j.comppsych.2018.03.004. - Maurage, P., Bestelmeyer, P.E.G., Rouger, J., Charest, I., Belin, P., 2013a. 2012). In addition, plasma levels of leptin, total and acety- Binge drinking influences the cerebral processing of vocal affective bursts lated, active ghrelin were measured prior to the fMRI ses- in young adults. NeuroImage Clin. 3, 218–225. https://doi.org/10.1016/j. sion after overnight fasting. Additionally, relapse data was nicl.2013.08.010. - Maurage, P., Pesenti, M., Philippot, P., Joassin, F., Campanella, S., 2009. collected during the three months following the assessment Latent deleterious effects of binge drinking over a short period of time using a semi-structured interview, which incorporated the revealed only by electrophysiological measures. J. Psychiatry Neurosci. 34, Alcohol Timeline Followback questionnaire (Sobell et al., 111. 1996). Brain imaging data were preprocessed and analyzed - Maurage, P., Petit, G., Campanella, S., 2013b. Pathways to alcohol-induced brain impairment in young people: A review by Hermens et al., 2013. using the statistical parametric mapping software for Matlab Cortex. 49, 1155–1159. https://doi.org/10.1016/j.cortex.2012.12.015. (SPM), according to standard procedures and previous stu- - Rupp, C.I., Derntl, B., Osthaus, F., Kemmler, G., Fleischhacker, W.W., 2017. dies (Bach et al., 2015). Associations between hormone levels Impact of social cognition on alcohol dependence treatment outcome: Poorer facial emotion recognition predicts relapse/dropout. Alcohol. Clin. and mesolimbic cue-reactivity were tested using multivariate Exp. Res. 41, 2197–2206. https://doi.org/10.1111/acer.13522. regression models in SPM, using a combined voxel- and - Sanhueza, C., García-Moreno, L.M., Expósito, J., 2011. Weekend alcoho- cluster-extent threshold that corresponds to a family wise lism in youth and neurocognitive aging. Psicothema. 23, 209–214. error rate correction of pFWE<0.05. Cox regression analyses - Stephens, D.N., Duka, T., 2008. Cognitive and emotional consequences of binge drinking: role of amygdala and prefrontal cortex. Philos. Trans. R. were performed to assess the associations between leptin, Soc. B. Biol. Sci. 363, 3169–3179. https://doi.org/10.1098/rstb.2008.0097. acylated ghrelin and relapse risk during the three months - Wills, T.A., Simons, J.S., Sussman, S., Knight, R., 2016. Emotional self- following the experiment. Results: analyses of psychometric control and dysregulation: A dual-process analysis of pathways to exter- nalizing/internalizing symptomatology and positive well-being in younger data showed that leptin plasma levels were negatively corre- adolescents. Drug. Alcohol. Depend. 163, S37–S45. https://doi.org/10.1016/j. lated with the scores of the Obsessive Compulsive Drinking drugalcdep.2015.08.039. Scale (r = -0.305, p = 0.020, pFDR = 0.040). For ghrelin, there was a positive association between acylated ghrelin levels and Opposing effects of the hormones leptin and changes in the intention to drink alcohol, such that higher ghrelin on neural alcohol cue-reactivity, craving acylated ghrelin levels were associated with an increase in the and relapse in alcohol addiction: two streams intention to drink alcohol (r = 0.331, p = 0.006, pFDR = 0.024). merge to one river? Multiple regression analyses in SPM showed a signifcant Patrick Bach1,2, Jan Malte Bumb1,2, Rilana Schuster1,2, Sabine negative association between leptin plasma levels and alco- Vollstädt-Klein1,2, Iris Reinhard1, Marcella Rietschel1, Stephanie hol cue-induced activation as the dependent variable in left H Witt1, Klaus Wiedemann3, Anne Koopmann1,2, Falk Kiefer1,2 (77.2% of cluster) and right caudate (18.3% of cluster), with a (1 Mannheim, Germany, 2 Heidelberg, Germany, 3 Hamburg, relevant proportion being located in the dorsal striatum (20.6 Germany) %), while only a small proportion was located in the ventral striatum (5.1%, combined threshold, corresponding to pFWE < Introduction: increasing evidence supports the role of appe- 0.05). In addition, mean alcohol cue-induced activation tite-regulating hormones in the pathophysiology of alcohol extracted from bilateral caudate, negatively correlated with addiction. Amongst those, leptin and ghrelin and a “cross- plasma leptin levels (r = -0.316, p = 0.016, pFDR = 0.040), cor- talk” between both hormones were implicated in the patho- roborating the results of the whole brain analyses. In contrast, of alcohol addiction, both modulating alcohol acylated ghrelin showed a signifcant positive association to craving and drug-seeking (Hirth et al., 2016). Preclinical alcohol cue-induced activation in several clusters of brain and clinical data thus far indicate that leptin and ghrelin areas, including the bilateral insulae and parts of the superior interact with each other and that both modulate the signaling and middle frontal gyri, as well as the middle cingulum. rate of dopaminergic neurons in reward networks (Fulton et Further, the mean functional activation in the left and right al., 2006; Farooqi et al., 2007; Haass-Kofer et al., 2015). insula signifcantly correlated with acylated ghrelin levels However, their role in alcohol addiction is far from being (r = 0.279, p = 0.013, pFDR = 0.026). Cox regression analyses understood, especially the neurobiological underpinnings of showed a signifcant association between leptin and time to their efects remains to be elucidated. To address this issue, heavy-relapse, such that high leptin levels during the post-

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cue reactivity and attentional bias in alcohol-dependent patients. Addict acute phase of withdrawal were associated with a longer time Biol, 17, 807-816. to frst heavy-relapse (Chi2 overall model = 4.308, Hazard Ratio = 0.922, 95%CI 0.853-0.996, p = 0.039), while acy- Altered GABA-signaling in the amygdala lated ghrelin and BMI did not contribute to the prediction contributes to pathological alcohol choice over of time to heavy-relapse (p > 0.684). Conclusion: we could high value alternative rewards show that leptin and acylated ghrelin showed opposing asso- Eric Augier1 (1 Linköping ,Sweden) ciations with the extent of alcohol cue-induced mesolimbic cue-reactivity and alcohol craving. Our fnding of a negative association between leptin and cue-reactivity in the bilateral Introduction: alcohol addiction is characterized by a progres- caudate and striatum is in line with previous evidence that sive shift of decision making, in which alcohol is increasingly supplementation of leptin attenuates mesolimbic hyper-acti- chosen over healthy non-drug rewards. Only a subset of vation in the NAc, caudate and putamen of leptin-defcient people transition from recreational to addictive alcohol use, a patients (Baicy et al., 2007; Farooqi et al., 2007). Te present pattern that is similar to that of other addictions. By contrast, results also mirror fndings of animal studies showing that in commonly used animal models, nearly all rats learn to leptin modulates fring of dopaminergic neurons in the VTA self-administer addictive drugs, including alcohol and animals that project to the striatum (Fulton et al., 2006). We thus have no alternative to drug use. Tese models fail to capture suspect that the reduced striatal cue-reactivity might be the an important and common feature of human addiction: a neurobiological correlate of leptin’s efect on relapse-risk. Te continued use of the substance despite the opportunity to fndings of a positive association between acylated ghrelin engage in important and meaningful social and recreational and cue-induced brain response in the left and right insula, activities. Purpose: the neurobiological underpinnings of harmonize with previous studies that showed that intrave- choosing alcohol over a natural reward in the context of nous administration of ghrelin to healthy volunteers during developing alcoholism are presently unknown. Here, we set an fMRI food-cue task, increased brain response in the out to identify molecular mechanisms underlying this choice amygdala, orbitofrontal cortex, insula, and striatum (Malik behavior. Methods: we frst employed an exclusive choice- et al., 2008), further supporting the plausibility of BOLD based method to identify rats that continue to self-administer response modulation by ghrelin. Te reported results further alcohol at the expense of a high-value natural reward, a sweet support the relevance of appetite regulating hormones in solution, and assessed whether these animals show other the pathophysiology of addiction and their potential role as characteristics reminiscent of clinical alcoholism. Specifcally, future treatment targets. we measured their motivation to obtain and take the drug References (modeled using an elevated breakpoint on a progressive-ratio - Bach, P., Vollstädt-Klein, S., Kirsch, M., Hoffmann, S., Jorde, A., Frank, J., schedule) and their continued drug use despite its harmful Charlet, K., Beck, A., Heinz, A., Walter, H., Sommer, W.H., Spanagel, R., and negative consequences (here modeled using continued Rietschel, M. & Kiefer, F. (2015) Increased mesolimbic cue-reactivity in car- alcohol self-administration despite quinine adulteration riers of the mu-opioid-receptor gene OPRM1 A118G polymorphism predicts drinking outcome: a functional imaging study in alcohol dependent sub- or delivery of a shock punishment contingent with drug jects. Eur Neuropsychopharmacol, 25, 1128-1135. delivery). We then carried out a discovery efort using gene - Baicy, K., London, E.D., Monterosso, J., Wong, M.L., Delibasi, T., Sharma, expression profling. Results: using this procedure in outbred A. & Licinio, J. (2007) Leptin replacement alters brain response to food cues in genetically leptin-deficient adults. Proceedings of the National Academy rats, we were able to identify two populations characterized of Sciences of the of America, 104, 18276-18279. by distinct patterns of choice behavior. Te vast majority of - Farooqi, I.S., Bullmore, E., Keogh, J., Gillard, J., O’Rahilly, S. & Fletcher, rats stop responding for alcohol when ofered the opportunity P.C. (2007) Leptin regulates striatal regions and human eating behavior. to access a high-value alternative reward (SP, Saccharin- Science, 317, 1355. - Fulton, S., Pissios, P., Manchon, R.P., Stiles, L., Frank, L., Pothos, E.N., preferring). However, a subpopulation (15% across multiple Maratos-Flier, E. & Flier, J.S. (2006) Leptin regulation of the mesoaccumbens batches of outbred Wistar rats, a proportion similar to human dopamine pathway. Neuron, 51, 811-822. alcohol addiction rates) continues to choose alcohol despite - Haass-Koffler, C.L., Aoun, E.G., Swift, R.M., de la Monte, S.M., Kenna, G.A. & Leggio, L. (2015) Leptin levels are reduced by intravenous ghrelin admi- the presence of the alternative (AP, Alcohol-preferring). nistration and correlated with cue-induced alcohol craving. Translational Rats that choose alcohol display a constellation of behavio- psychiatry, 5, e646. ral traits reminiscent of clinical alcoholism, mimicking key - Hirth, N., Meinhardt, M.W., Noori, H.R., Salgado, H., Torres-Ramirez, O., clinical diagnostic criteria for alcoholism. Specifcally, they Uhrig, S., Broccoli, L., Vengeliene, V., Rossmanith, M., Perreau-Lenz, S., Kohr, G., Sommer, W.H., Spanagel, R. & Hansson, A.C. (2016) Convergent show elevated motivation to obtain alcohol, as measured by evidence from alcohol-dependent humans and rats for a hyperdopaminer- progressive ratio breakpoints and continue to take alcohol gic state in protracted abstinence. Proceedings of the National Academy of despite negative consequences that make the majority of rats Sciences of the United States of America, 113, 3024-3029. - Malik, S., McGlone, F., Bedrossian, D. & Dagher, A. (2008) Ghrelin modu- stop self-administration, namely adulteration with increasing lates brain activity in areas that control appetitive behavior. Cell Metab, 7, concentrations of the bitter tastant quinine, or pairing of the 400-409. alcohol delivery with foot-shock. Furthermore, a diferential - Sobell, L.C., Brown, J., Leo, G.I. & Sobell, M.B. (1996) The reliability of the gene expression screen using a custom NanoString nCounter Alcohol Timeline Followback when administered by telephone and by com- puter. Drug and alcohol dependence, 42, 49-54. panel found minimal evidence for diferential gene expression - Vollstadt-Klein S, L.S., Richter A, Kirsch M, Bach P, von der Goltz C, between alcohol choosing vs non-choosing rats in several Hermann D, Mann K, Kiefer F. (2012) Validating incentive salience with brain structures examined, but did identify that a marked functional magnetic resonance imaging: association between mesolimbic dysregulation of a GABAergic pathway within central amyg-

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 236 Congress dala (CeA) was associated with addiction-like phenotype. to date (i.e., CGP7930, GS39783, BHF177, rac-BHFF, A > 50% down-regulated expression of the g-aminobutyric CMPPE, ADX71441, COR659, and ORM-27669) have acid (GABA) transporter GAT-3 (Slc6a11) in this structure invariably been reported to reduce, or even suppress, several was accompanied by similarly down-regulated transcripts alcohol-motivated behaviors in rats and mice. Tese beha- encoding several GABAA receptor subunits. Tis indicated viors include excessive alcohol drinking, alcohol relapse-like the possibility of elevated inhibitory GABA-tone due to drinking, alcohol binge-like drinking, operant oral alcohol impaired clearance of extracellular GABA by the transporter, self-administration (under both fxed and progressive ratios a hypothesis that was confrmed using slice electrophysiology. of reinforcement), reinstatement of alcohol seeking, alcohol- Additionally, a viral-vector mediated knockdown of GAT- induced locomotor stimulation, and alcohol-induced condi- 3 resulted in increased GABA-mediated inhibition in the tioned place preference. Te use of validated animal models of CeA in a slice electrophysiology experiment, and converted several aspects of AUD confers remarkable translational value SP rats into AP rats in vivo, demonstrating a causal role to these fndings. Among these GABAB PAMs, COR659 of GAT-3 for alcohol choice. Finally, we assessed whether appears to be of particular interest because of a likely dual these results may have translational relevance and carried out , involving – beside the positive allos- an RNAseq transcriptome analysis in post-mortem tissue teric modulation of the GABAB receptor – an action at the from alcohol dependent people and controls. We found that cannabinoid CB1 receptor. Tis results in a unique behavioral GAT-3 expression was selectively decreased in the central pharmacological profle, including suppression of multiple amygdala of alcohol-dependent people compared to those behaviors motivated by alcohol and food (including highly who died of unrelated causes. Conclusion: our data provide palatable food) in rats. To summarize, data collected to date strong support for a causal contribution of neuroadapta- (i) confrm that the GABAB receptor is a major part of the tions afecting GABA signaling within the amygdala to the neural substrate controlling alcohol drinking and mediating development of alcohol addiction. Furthermore, our fndings the reinforcing, motivational, stimulating, and rewarding pro- suggest that pre-existing diferences in GABAergic gene perties of alcohol, and (ii) suggest that positive modulation expression in the CeA may also infuence susceptibility to of the allosteric binding site(s) is an efective mechanism, developing alcohol addiction. Collectively, these experiments in addition to activation of the orthosteric binding site, to identify that impaired GABA clearance within the amygdala potentiate GABAB receptor-mediated neurotransmission and contributes to alcohol addiction, appears to translate between inhibit alcohol-motivated behaviors. Te recent transition of species, and may ofer targets for new pharmacotherapies for the frst GABAB PAMs to the initial steps of clinical testing treating this disorder. makes investigation of efcacy of GABAB PAMs in AUD patients an important and feasible option. GPCRs & alcohol-seeking Loss of control over alcohol drinking behaviour is linked to persistent changes in the dopaminergic Andrew Lawrence (Melbourne, Australia) and systems Valentina Vengeliene1, Rainer Spanagel2, Anita C Hansson2 ((1 Vilnius, Lithuania, 2 Mannheim, Germany) New lines of experimental evidence on the reducing effects of positive allosteric modulators of Repeated exposure to deprivation phases in long-term alcohol the GABAB receptor on alcohol-motivated behaviors drinking Wistar rats has proven to be a useful method to Giancarlo Colombo1, Paola Maccioni1 (1 Monserrato, Italy) induce addictive features, such as loss of control over drinking behaviour. In this study we explored neurobiological mecha- Among G-protein-coupled receptors, the GABAB recep- nisms that underlie transition from controlled to compulsive tor has recently gained considerable interest in the alcohol alcohol consumption. For this purpose male rats were given research feld: the prototypic GABAB receptor , baclo- concurrent ad libitum access to water, as well as to 5%, 10% fen, has repeatedly been reported to suppress several alcohol- and 20% alcohol solutions during an observation period of related behaviors in laboratory animals as well as alcohol eight months. Loss of control over drinking behaviour was consumption and craving for alcohol in patients afected measured during post-abstinence drinking phase by adding by alcohol use disorder (AUD). More recently, preclinical quinine hydrochloride to alcohol solutions and by paring research in the alcohol feld has focused on the positive allos- mild foot-shock with instrumental responding for alcohol. teric modulators (PAMs) of the GABAB receptor, a new class Changes in mesocorticolimbic and nigrostriatal neurotrans- of GABAB receptor agents with improved safety profle in mission induced by long-term alcohol consumption were comparison to baclofen. Te present talk will summarize the assessed by means of receptor autoradiography for dopamine several lines of experimental evidence unanimously indicating and opioid receptor in rats withdrawn from alcohol for three that GABAB PAMs retain the ability of baclofen to inhibit weeks. Our results showed that loss of behavioural fexibility multiple alcohol-motivated behaviors in rodents; importantly, in voluntary alcohol drinking rats are, at least partly, caused the efects of GABAB PAMs are selective for alcohol and oc- by persistent upregulation of µ-opioid receptor, downregula- cur at doses largely lower than those producing hypolocomo- tion of dopamine transporter and upregulation of dopamine tion and sedation. More specifcally, all GABAB PAMs tested receptor D1 in the ventral striatum and mPFC. Dopamine

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 237 Congress receptor D2 levels were unafected by long-term drinking alcohol seeking to aid identifying and evaluating novel tar- procedure. Our fndings suggest that development of addic- gets. AUDs are characterised by a transition to compulsive tive behaviour could be a result of lost ability of the brain alcohol seeking, which is hypothesized to involve a shift from to adapt to a changing environment with respect to alcohol ventral to dorsal striatum. In addition, a medial to lateral removal and re-exposure, which points at the importance of shift in the dorsal striatum is implicated in the transition reversal of the lost brain plasticity in the development of new from goal-directed to habitual alcohol seeking. Muscarinic treatment strategies. and nicotinic acetylcholine receptors (AChRs) are potential targets for AUD treatment as they are expressed within the The role of /orphanin FQ NOP receptor mesocorticolimbic reward system, including dense expression system in alcohol abuse 1 1 1 in the dorsal striatum. Here they modulate dopamine and Roberto Ciccocioppo , Anna Maria Borruto , Yannick Fotio , glutamate release, which may regulate reward processing. 2 1 1 1 Friedbert Weiss , Alice Ilari , Michele Petrella , Alessio Masi , To assess the role of AChRs in AUD, we frst conducted 1 1 2 Nazzareno Cannella ( Camerino, Italy, La Jolla, USA) genome-wide RNA sequencing in the caudate/putamen of 10 human alcoholics and 10 healthy controls and concurrently Te 17 amino acid Nociceptin/Orphanin FQ (N/ examined AChR expression in the corresponding regions in OFQ) is the natural ligand for the orphan G protein-cou- rat (dorsolateral and dorsomedial striatum) following chro- pled receptor (GPCR) Opioid Receptor Like-1 (ORL1), nic alcohol consumption/withdrawal using qPCR. Next we now known as NOP. Earlier studies in genetically selected examined the role of select mAChR and nAChR subtypes Marchigian Sardinian (msP) alcohol preferring rats and in in alcohol consumption and seeking using selective allosteric alcohol dependent Wistars demonstrated that activation modulators. Finally, we probed the role of select mAChR and of NOP by selective ligands reduced alcohol drinking and nAChR subtypes in the dorsal striatum in alcohol consump- seeking. Te efect was particularly pronounced following tion and seeking. Collectively, our data show that specifc repeated drug administration. More recently, using selective mAChRs are potential novel target pharmacotherapies for NOP antagonists our laboratory found that NOP receptor the treatment of AUD. blockade also reduced alcohol intake and relapse elicited by stress or by cues predictive of substance availability. Moreover, in a series of experiments aimed at characterizing the neuro- Cellular and extracellular effects of alcohol circuitry mediating the efects of these drugs we found that exposure in liver and extrahepatic organs the central amygdala (CeA) plays a critical role in the efect of NOP . Whereas, the efects of the antagonists involve Carol Casey (Omaha, USA) both the CeA and the ventral tegmental area (VTA). Why both, NOP agonists and antagonists, reduces the motivation for alcohol is still unclear. However, based on evidence that TP-R deletion promotes adipose tissue browning NOP agonists like Ro 64-61 98 and MT-7716 reduced alco- and M2 macrophage polarization with a hol drinking following chronic but not acute administration concomitant reduction in alcohol-associated liver we propose that multiple dosing of NOP agonists, through injury in mice receptor desensitization, may reduce N/OFQ transmission. 1 1 1 Tis hypothesis is corroborated by binding data showing that Saraswathi Viswanathan , Terrence Donohue , Carol Casey 1 in msP rats repeated injections of Ro 64-6198 down-regulated ( Omaha, USA) the expression of NOP receptors in various brain areas. Based on data showing that NOP receptors are upregulated in Ethanol (EtOH) administration leads to changes in adipose high alcohol drinking animal like msP and in postdependent tissue (AT) lipid storage function and metabolism. In a Wistars we propose a heuristic model according to which model of non-alcoholic fatty liver disease (NAFLD), mice increased activity of N/OFQ system facilitate alcohol use. Tis lacking thromboxane-prostanoid receptor (TP-R) were view is corroborated by data showing that rats with genetic protected against NAFLD with a concomitant increase in deletion of NOP receptors drink less alcohol compared to markers of AT browning. Here, we hypothesized that ethanol wild type control Grant support (AA017447 and AA014351). would induce AT lipolysis to promote AT browning and that blocking TP-R would enhance EtOH-induced AT brow- Muscarinic acetylcholine receptors in alcohol use ning and attenuate free fatty acid (FFA) fux to liver which disorder would ameliorate alcohol-associated liver disease (AALD). 1 1 1 Aute Leigh C Walker , Alice E Berrizi , Nicola A Chen , Victoria Male TP-R knock out (KO) and wild type (WT) littermate 1 1 2 2 Perreau , Patricia Rueda , Craig W Lindsley , Carrie K Jones , controls were fed Lieber-DeCarli control or EtOH liquid Christopher J Langmead1, Andrew J Lawrence1 (1 Melbourne, diets for 4 wk. Chronic EtOH feeding increased mRNA for Australia, 2 Nashville, USA) Ucp-1, a marker of AT browning, in visceral AT (VAT) of WT mice (P<0.05) which was further elevated in TPR-KO Despite the large socioeconomic burden of alcohol use disor- mice (P<0.01). Moreover, we found a remarkable increase in ders (AUD), therapeutic treatment options are limited. Tere 3-adrenergic receptor (Adrb3) mRNA expression, which is a need to characterize the neurochemistry underpinning stimulates AT browning, in VAT of KO mice but not in WT

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 238 Congress mice after EtOH exposure (P<0.01). RNAs encoding M2 (CCl4) in combination with chronic intra-gastric ethanol anti-infammatory macrophages were signifcantly increased feeding, they develop an alcoholic liver fbrosis-associated in EtOH fed-KO, but not WT mice. On the other hand, AKI (10.1016/j.taap.2016.09.011). Using publically available mRNA levels of Ccl2 (encoding MCP-1), an infammatory data, we found that 23 of the 1996 diferentially expressed marker, rose signifcantly in EtOH-fed WT, but not KO genes in AKI mice were complement genes (GSE83529). mice. Plasma MCP-1 levels followed this same trend. Our Further, when DEGs were fltered into the protein-protein data show that blockade of TP-R enhances the efect of interaction (PPI) network complex, 8 of the top 30 hub EtOH in inducing browning markers in AT. Additionally, DEGs were found to be complement genes. We also fnd TP-R deletion leads to AT macrophage polarization and evidence of complement activation in liver explants from an M2 anti-infammatory phenotype. Tus, blocking TP-R patients with AH compared to healthy controls (tissue function in AT may be efective in ameliorating AALD. from the Clinical Resources for AH Investigations at Johns Hopkins University). Further, the concentrations of C5a, a Down-regulation of Rab3D: critical role in alcohol- potent anaphylatoxin, and factor Ba, an indicator of alterna- induced liver injury tive pathway activation, were elevated in both the circulation 1 1 1 Carol Casey , Karuna Rasineni , Terrence Donohue , Paul and urine from patients with AH from the DASH consor- 1 1 1 Thomes , Armen Petrosyan ( Omaha, USA) tium compared to healthy controls. Kidney Injury Marker-1 (KIM-1) increased in urine of severe AH patients compared Te goal of our current work is to examine how ethanol to moderate AH. Taken together, these data suggest that exposure results in impaired function of the Golgi apparatus. complement activation is associated with AKI in AH and Te Golgi apparatus (also called the Golgi body or Golgi inhibition of complement activation may be a potential target complex) packages proteins into membrane bound vesicles for clinical intervention. inside the cell before the vesicles are sent to their destination. Masked hemolysis as important factor of iron As such, this organelle resides at the intersection of the secre- overload in ALD tory, lysosomal, and endocytic pathways; it is known to be of 1 1 1 particular importance in processing proteins for secretion. Vanessa Rausch , Inês Silva , Teresa Peccerella , Sebastian 1 1 Previous work from our laboratory has identifed multiple Mueller ( Heidelberg, Germany) defects in endocytosis, protein trafcking, and secretion, after alcohol administration, but we have not until now, examined Background: 50% of ALD patients develop hepatic iron over- a role for altered Golgi function in these processes. Of cen- load (HIO) and anemia, however, the underlying mechanisms tral importance to our study is the role of a small GTPase, including hepcidin response are poorly understood. Herein, Rab3D, which is involved in exocytosis, secretion and vesicle we introduce hemolysis as novel factor in disrupting hepcidin trafcking. We have shown that Rab3D protein content regulation and eventually causing iron overload. Method: we was signifcantly decreased after alcohol administration, and here studied hepcidin, molecular and laboratory iron markers recently we have obtained exciting new preliminary data that in ALD patients (n=831, mean alcohol consumption 192 g/ ethanol-impaired Rab3D function plays an important role in day). Te efect of hemolysis was further studied in C57BL/6 Golgi disorganization and fragmentation. We are focusing mice using phenylhydrazine (PHZ)-induced hemolysis our examination on a role for Rab3D in transport through model. Finally, in vitro erythrophagocytosis model was used the Golgi, and have data showing how alcohol-induced to recapitulate in vivo fndings and to investigate the under- remodeling of Golgi morphology is a signifcant impairment lying mechanisms. Results: indirect evidence for hemolysis of post-Golgi trafcking, and may lead to utilization of trans- (anemia, ferritin, LDH, MCV, CD163) as cause for HIO Golgi membranes for the formation of autophagosomes. was found in 16.4% of heavy drinkers. Despite higher ferritin With our expertise and experimental models, we hope to levels as compared to controls, hepcidin was not adequately provide novel insights as to how alcohol afects trafcking of upregulated in hemolytic patients suggesting a suppressive proteins within the liver cells leading to toxicity. effect. In confirmation, evere PHZ-induced hemolysis (anemia, transaminases, LDH) suppressed hepcidin in mice Complement activation in alcoholic hepatitis: leading to anemia, elevated transaminases and transferrin possible association with acute kidney injury saturation and LDH levels. Phagocytosed erythrocytes were Laura E Nagy1 (1 Cleveland, USA) detected in the spleen and iron-loaded Kupfer cells in the liver. In vitro, erythrophagocytosis led also to the suppres- Complement contributes to liver injury in murine models of sion of hepcidin at higher pathological levels of oxidized alcoholic liver disease (ALD); however, the role of comple- erythrocytes. Conclusion: our data suggest that suppression ment in patients with alcoholic hepatitis (AH) is not well of hepcidin by masked hemolysis seems to be an important studied. Acute Kidney Injury (AKI) is a major predictor of factor contributing to HIO in ALD patients. mortality in severe AH. Since complement activation contri- butes to AKI in other kidney diseases, here we investigated Second hits in alcohol-related organ damage the association between complement activation and AKI in ALD, making use of both murine models and patient samples. When mice are exposed to carbon tetrachloride Kusum Kharbanda (Omaha, USA)

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 239 Congress

Alcohol on fat: worse consequences for liver injury Using ROC analysis, LS was the best predictor of death in 1 1 1 1 E. Huang , A.M.P. Duly , C. Yee , S.V. McLennan , Devanshi general with an AUROC of 0.72 and a cutof value of 14.0 Seth1 (1 Sydney, Australia) kPa, followed by AP and albumin. Moreover, LS was the top predictor of death starting from 2 to 5 years. In contrast, Background and aims: drinkers who are obese are more likely LS was preceded by bilirubin and albumin in predicting to develop liver cirrhosis than those within a healthy weight one-year-survival. Conclusion: we here identify LS as the range implying the potential for an interaction between best long-term prognostic parameter in patients who hea- alcohol and obesity. Experimental models of alcohol and high vily consume alcohol. LS measurements should become an fat diet (HFD) alone have proven difcult to induce severe important parameter for the screening of alcoholics. liver injury even after several weeks of treatment. LPS is Reactive aldehydes from alcohol and cigarette commonly required as a “second hit” with alcohol to advance smoke co-exposure impairs lung innate anti- steatosis to steatohepatitis and in diet-related obesity models induction of diabetes accelerates liver injury. We studied the microbial defense 1 1 interaction between alcohol and HFD in liver injury mouse Todd A Wyatt ( Omaha, USA) model comparable to human setting of episodic heavy drin- king with fat-rich food. Methods: C57BL6 male mice were Te vast majority of individuals with an alcohol use disorder fed either chow or high fat diet (HFD) ad libitum for 12 smoke cigarettes. Alcohol misuse and cigarette smoking are weeks. A sub-set of mice from each group were also given co-morbidities resulting in the signifcant susceptibility to alcohol (2g/kg body weight) twice/week via intra-gastric lung infections leading to pneumonia. Several innate lung lavage. Liver injury was examined by histopathology and defense mechanisms are altered by the combination of liver/serum biochemistry. Expression of molecules related to drinking alcohol and smoking cigarettes. Mucociliary clea- lipid metabolism, infammation and fbrogenesis were exami- rance is negatively impacted by smoke and alcohol through ned (Q-PCR, immunofuorescence). Results: we show that cilia slowing and ciliated cell detachment caused by the chronic moderate alcohol exacerbates liver injury in a mouse co-exposure-induced activation of protein kinase C epsilon. model of HFD compared to either alcohol or HFD alone. Malondialdehyde and acetaldehyde are generated via both Alcohol superimposed on HFD increased serum and liver alcohol metabolism as well as the pyrolysis of tobacco leading lipids, triglycerides, cholesterol, circulating , infam- to the formation of stable hybrid adducts known as MAA mation and several molecules related to lipid processing. In adducts. Lung surfactant protein is altered through MAA addition, immune cells also increased with Alcohol+HFD. adduction resulting in decreased anti-microbial action by the Whereas alcohol alone moderately increased ethno-adducts, collectin. Lastly, the secretion of lung mucosal immunoglo- Alcohol+HFD fed animals showed a striking elevation of bulin A is decreased through the action of MAA adducts in etheno-DNA adducts clusters within the liver suggesting a a TGF beta-dependent manner. Both airway epithelial cells precancerous profle in our model. Conclusion: alcohol on fat and macrophages bind MAA adducted protein via CD204 worsens liver injury. suggesting that diferential expression of CD204 polymor- phisms may govern injury due to co-exposure. Tus, the Liver stiffness as a novel prognostic marker in suppression of lung innate defense is multi-faceted in alcohol heavy drinkers: first data from a prospective misuse due to the comorbidity of cigarette smoking. 10-year follow-up study Sebastian Mueller1 (1 Heidelberg, Germany) Matrix stiffness regulates fibrosis progression in alcohol-induced liver injury Background and aims: we here present frst data on the pro- Senthilkumar Thulasingam, Michael Moeller, Madhusudanan gnostic impact of liver stifness (LS) on long-term survival of Narasimhan, Carol Casey, Srivatsan Kidambi1 (1 Lincoln, USA) Caucasian heavy drinkers in a 10 year, prospective single cen- ter trial. Method: information of survival status was obtained Liver stifness (LS) is widely used clinically to monitor, sta- in 675 (71.6%) of 943 screened patients that had presented ging and diagnosis, of alcoholic fatty liver diseases (AFLD) for alcohol detoxifcation over a 10-year period from 2007- and is also believed to predict improved outcome. Te regu- 2017 with a mean daily consumption of alcohol of 178 g. latory mechanisms of LS in regulating liver function during Mean observation time was 3.7 years and mean duration of ALD, specifcally, fbrosis, is incompletely understood. Tis heavy drinking was 14.0 years. All patients had LS measure- study aims to uncover signifcant mechanisms underlying the ments by transient elastography and routine laboratory tests. role of increasing matrix stifness during liver injury in the Results: during the observation time, 106 patients (15.7%) promoting fbrogenesis induced by alcohol. We hypothesize died. Te cause of death could be clarifed in 42 patients that LS is just not the readout of fbrosis but also an active (39%) and it was liver-related in 16 (38%). Overall death was contributor of the progression of liver fbrosis and stellate cell highest associated with LS (r=0.291, P=1.3E-14), followed by activation in AFLD. Using our innovative biomimetic liver hemoglobin and alkaline phosphatase (AP). In a multivariate fbrosis model that allows modulation of substrate stifness proportional hazard model, LS next to age, AP and serum (2 kPa, 9 kPa 25 kPa and 55 kPa mimicking healthy, early albumin was the most signifcant independent predictor of fbrotic, fbrotic and extremely fbrotic substrates), we investi- survival with a hazard ratio of 1.013 (1.003 to 1.023, P<0.05). gated the role of liver matrix stifness in modulating primary

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 240 Congress hepatocytes and stellate cell function. In vitro experiments interplay; particularly regarding tobacco use, whatever the were designed using the conditioned medium (CM) of pri- underlying mechanisms. Future studies should investigate mary hepatocytes (isolated from alcohol fed rats) cultured on mechanisms related to the observed associations. stifness mimicking healthy and fbrotic environment supple- Development of a predictive clinical tool of mented to human stellate cells (HSC). A signifcant increase alcohol-related consequences: results from the in HSC proliferation, and expression of fbrosis-related genes Epidemiologic Survey on Alcohol and Related were observed in cells treated with CM from stifer matrix. Together, all these data demonstrates the plausible role of Conditions 1 stifness in regulating hepatocytes function and contribute to Nicolas Hoertel , Marie Dosquet, Hugo Peyre, Carlos Blanco, stellate cells activation and progression of liver fbrosis during Géraldine Ducoutumany, Philip Gorwood, Henri Leleu, alcohol liver disease. Understanding the impact of stifness on Guillaume Airagnes, Cédric Lemogne, Henri-Jean Aubin, hepatocytes biology will provide signifcantly more nuanced Frédéric Limosin (1 , France) data to aid drug development for AFLD and liver fbrosis. Objective. To develop an individualized risk calculator tool of the 3-year risk of 6 important alcohol-related medical, psy- What can we learn from in chological and social adverse outcomes based on predictor va- alcohol research? Recent findings from large riables that are easily and routinely collected in primary heal- population-based cohorts thcare settings. Material. A nationally representative sample of US adults aged 18 years or older was interviewed 3 years apart in the National Epidemiologic Survey on Alcohol and Guillaume Airagnes (Paris, France) Related Conditions (wave 1, 2001-2002; wave 2, 2004-2005). Analyses concerned 22,009 respondents interviewed in both Do addictive behaviors explain social inequality waves and using alcohol in wave 1. Tis sample was randomly regarding depression? Findings from the split into a construction (N=11,013) and a validation sample Constances cohort (N=10,996). Methods. Te 3-year risk of 6 important alcohol- related adverse outcomes (i.e., alcohol use disorder, with- Joane Matta1, Nicolas Hoertel, Guillaume Airagnes, Emmanuel drawal symptoms, occurrence of tremors or seizures, interper- Wiernik, Frédéric Limosin, Marcel Goldberg, Marie Zins, Cédric 1 sonal relationship problems and legal problems) was modeled Lemogne ( Villejuif, France) in the construction sample using logistic regression, with age, sex and the 3 AUDIT-C variables as predictors. Scoring was Objective. To examine the associations between depression used to combine information derived from predictors and and alcohol, tobacco and cannabis, taking into consideration quantify alcohol-related risks for each subject. Discrimination socioeconomic status (SES). Methods. We applied media- and calibration were assessed in the validation sample based tion and moderated mediation models stratifed for sex to on the C-index and the Hosmer and Lemeshow (H-L) test. a nationally representative sample (N=37,192) of French Results. Te predictive values of the risk equations were good men and women from the Constances cohort with baseline (C-index ranging from 0.75 to 0.83) and calibrated well (all and follow-up measures regarding depressive symptoms. Te H-L test p-values>0.44) in the validation sample, showing structural equation model tested the associations between low potential clinical usefulness. Conclusions and Relevance. SES status (income and education, separately) and depressive This clinician-friendly individualized risk calculator can symptoms at follow-up mediated by alcohol, smoking and be useful to identify individuals with a short-term risk of cannabis, while taking into consideration age and depres- developing alcohol-related adverse outcomes, encourage sive symptoms at baseline. A second set of analyses tested at-risk drinkers to cut down their drinking and facilitate the mediation and moderation models with interactions the implementation of focused preventive interventions. between SES and substances. Results. Mediation analyses using low education or low income did not explain the Associations of depressive symptoms with alcohol association between SES and depressive symptoms in either use: findings from the Constances cohort 1 men or women. Mediation and moderation models showed Emmanuel Wiernik , Nicolas Hoertel, Guillaume Airagnes, that direct efects were not signifcant in the presence of Joane Matta, Frédéric Limosin, Marcel Goldberg, Marie Zins, interactions and that the moderation efect was largely signi- Cédric Lemogne (1 Villejuif, France) fcant. Direct associations between substances and SES or depressive symptoms were only retained for tobacco use. In Background. It remains unclear whether the risk of alcohol the low education models, the estimate of moderation was use is related to specifc depressive symptoms (e.g. poor appe- 0.278±0.076 and 0.182±0.09 in men and women respectively. tite), to specifc dimensions underlying depression (e.g. soma- Strong moderations efects were also found in the low income tic symptoms), to a general depression factor representing the models (0.348±0.076 and 0.247±0.08 in men and women, shared efect of all depressive symptoms, or to a combination respectively). Conclusion. Prevention strategies targeting at of these explanations. In addition, these efects could be risk subgroups should consider SES and substance use not specifc to alcohol or common to other substances. Methods. only as a cumulative risk factor but take into account their From 14,117 men and 14,629 women included from January

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 241 Congress

2015 to December 2016 in the French Constance cohort, we ral hostility predicted alcohol consumption over a 21-year applied structural equation modeling to examine the shared follow-up. Interventions aiming at modulating behavioral and specific effects of depressive symptoms (Center for hostility may help reducing its long-lasting infuence on Epidemiological Studies-Depression) on alcohol, tobacco, e- alcohol consumption. cigarette and cannabis use, while taking into account the co- occurrence between those substances. Analyses were stratifed by sex and adjusted for age and education. Results. Heavy Beyond cognition: neuroscience correlates of alcohol use was significantly associated with depression affective and motivational processes in binge and this association was mostly mediated through a general drinking depression factor (i.e. shared efect of all depressive symp- toms). Tis was also the case for the other substances. Beyond Pierre Maurage (Louvain-la-Neuve, Belgium) and above the efect of that general factor, reduced positive symptoms (e.g. ) had an additional efect on heavy alcohol use, particularly in men. Somatic symptoms had an The role of interoception and emotional additional efect on cannabis use. Conclusions. Because the impulsivity in binge drinking association between depressive symptoms and substance use Aleksandra Herman1 (1 Brighton, UK) was mainly mediated through a general depression factor, a better knowledge of biological and psychological mechanisms Despite our increasing knowledge of the cognitive as well underlying this dimension may help reduce the burden of as emotional alterations in binge drinking, the factors that substance use. In addition, the importance of particular subs- actually predispose to binge drinking remain unclear. More tance-specifc dimensions of depression could help to better recently the role of interoception, defned as the sense of the identify at-risk individuals. physiological state of the body, is being investigated in alco- Do personnality traits predict alcohol consumption hol use and misuse. Te talk will present our recent neuroi- decades after their assessments? Findings from maging as well as behavioural fndings focusing on the role of the Gazel cohort emotional impulsivity and interoception as well as emotional Guillaume Airagnes1, Cédric Lemogne, Alice Gueguen, Nicolas recognition alterations as driving factors for alcohol use in Hoertel, Marcel Goldberg, Frédéric Limosin, Marie Zins (1 Paris, non-dependent binge drinkers. A model will be presented of France) how these factors interact with each other in the context of identifying potential endophenotypes associated with risk for the development of alcohol addiction. Background. Hostility has been found to be positively associa- ted with alcohol intake in cross-sectional studies. Our aim was Emotional impairments in binge drinking: insights to examine prospectively the long-lasting association of hosti- through a behavioral and neuroscientific approach lity with alcohol consumption. Methods. We included 10,612 Severine Lannoy1 (1 Reims, France and Stanford, USA) men and 3,834 women from the French Gazel cohort with mean ages in 1993 of 48.6 (SD=2.9) and 45.7 (SD=4.2), res- Binge drinking is a widespread alcohol consumption pattern pectively. Hostility (i.e. total, cognitive and behavioral) was as- in young people. It has been related to deleterious conse- sessed in 1993 with the Buss and Durkee Hostility Inventory. quences such as brain modifcations and cognitive dysfunc- Alcohol consumption was self-reported annually from 1994 tions. Recent studies have also proposed that binge drinking to 2014. Hostility scores were introduced successively in gene- may be associated with emotional difculties. Te talk will ral linear mixed models with annual alcohol consumption in present behavioral, electrophysiological, and neuroimaging drinks per week as dependent variable. Multivariable analyses fndings supporting this proposal. It will underline the exis- were adjusted for age, occupational status, marital status, tence of emotional defcits in binge drinkers as well as the retirement status and depression score. All the analyses were heterogeneity of this population. Electrophysiological and stratifed by sex. Results. Among men(women), 83.0%(76.2%) neuroimaging results related to distinct paradigms involving completed at least 75% of all annual assessment of alcohol the processing of emotional stimuli will also be presented to consumption over a 21-year follow-up. In univariate analysis, extend the understanding of emotional difculties in binge alcohol consumption was associated with total and behavioral drinkers. Tese results reinforce the importance of emotional hostility in both sex (all p<0.001). In multivariable analyses, processes in binge drinking and open new research avenues these associations remained signifcant with a greater size in the feld of alcohol-use disorders. efect for behavioral hostility. Estimated means of alcohol consumptions ranged from 10.50[95CI%:10.01-10.92] drinks How do binge drinkers inhibit alcoholic images? A per week to 13.32[95%CI:12.90-13.74] in men and from functional magnetic resonance imaging study 4.09[95%CI:3.71-4.46] to 5.78[95%CI:5.39-6.17] in women, Sónia S Sousa1 (1 Braga, Portugal) for the frst and last quartiles respectively (p trends<0.001 and all pairwise comparisons<0.01). Similar efects were observed Binge Drinking (BD) is characterized by the consumption of among participants with at-risk alcohol consumption at large amounts of alcohol in a short time followed by a period baseline. Conclusions. In both men and women, behavio- of abstinence or very low consumption. Tis pattern of alco-

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 242 Congress hol consumption is highly prevalent among adolescents and immature cognitive control system. As a result, adolescence young adults, especially college students, and an important is characterized by elevated risk-taking behaviours and risk factor for substance abuse. Although BD is not conside- seeking of novelty and rewarding sensations, associated with red an addictive disorder per se, binge drinkers (BDs) display inefcient decision-making abilities. As the adolescent brain cognitive defcits similar to addicted individuals. Namely, is still under critical development, alcohol misuse has serious defcits in executive functions, such as in inhibitory control, deleterious efects, which may exacerbate the problem. To impulse control and delay of gratifcation, in addition to func- explore brain activity associated with inhibitory control in tional alterations of the frontal networks involved in addictive motivational alcohol-related contexts in young binge drin- behaviours. Tus, while behavioural diferences are not usually kers (BD), a sample of college students performed a Go/ observed between BDs and light social drinkers, increased NoGo task with beverage (alcohol vs. non-alcohol) stimuli activation in the BDs’ frontal or fronto-parietal regions while ERPs (80 controls, 71 BD) and fMRI (36 controls, 32 during the performance of executive-related tasks is relatively BD) were recorded. Diferences between BD and controls consistent across studies. Tis greater neural activation seems were found in the anterior N2 ERP wave and in the infe- to refect a brain compensatory mechanism that enables rior frontal cortex activity during response inhibition. Tese BDs to maintain task performance levels similar to controls. diferences were modulated by the alcohol-related content of However, current knowledge on the BDs’ neurofunctional stimuli. Te results will be discussed in the frame of the dual- response when inhibiting a motor response to alcoholic sti- process models, providing new evidence on the understanding muli is scarce. In this sense, the main purpose of the present of excessive drinking habits in youth. study was to assess functional activity in the brain networks associated with motor response inhibition to alcoholic stimuli in young BDs. Twenty College BDs and 16 age-matched Don’t stress the amygdala – the role of pro- and non-alcohol consumers (NACs) (18-23 years-old) underwent anti-stress systems in AUDs a functional magnetic resonance imaging (fMRI) acquisition while performed a stop signal task with alcohol-related and Sophia Khom (La Jolla, USA) non-alcohol-related stimuli. At the behavioral level, BDs exhibited lower reaction times than NACs in response to alcoholic stimuli. When analyzing neural activation, BDs dis- Corticotropin releasing factor binding protein in played increased activity over the right dorsolateral prefrontal the amygdala: the good, the bad and the ugly cortex during response inhibition to alcoholic stimuli. In Carolina L Haass-Koffler1 (1 Providence, USA) addition, BDs showed augmented activation in limbic regions such as the parahippocampal gyrus, when observing alcoholic To investigate the causal link between corticotropin releasing stimuli. Finally, brain activity of visual regions for alcoholic factor binding protein (CRFBP) in the central nucleus of the drinks was superior relative to non-alcoholic beverages in the amygdala (CeA) and alcohol-seeking behaviors, we utilized BD group only. Overall, our results suggest that BDs may a rat model trained to self-administer ethanol, designed with need additional cognitive resources to perform similar to controlled regional expression of CRFBP in the CeA and NACs when inhibiting a response to alcoholic stimuli. Tese electrophysiology data in transgenic mice expressing green fndings are in agreement with the only fMRI study conduc- fuorescence protein (GFP) in CRF Receptor 1 expressing ted to date on motor inhibition to alcoholic stimuli in BDs neurons in chronic ethanol exposure. In rats, we found that (Amesa et al., 2014), pointing to a compensatory neurofunc- CRFBP downregulation in the CeA reduces ethanol self- tional mechanism that may allow BDs to perform efciently administration and CeA hemodynamic activity but does not in inhibitory control-related tasks. Additionally, BDs seem to attenuate yohimbine-induced ethanol self-administration. be more impulsive in their responses to alcoholic stimuli and In mice, preliminary data suggest that CRFBP inhibition to be attentionaly biased towards this type of stimulus, which decreases postsynaptic GABAA receptor function through in turn could be activating emotional memories contributing CRF R2 in the CeA. All these data support our hypothesis to alcohol consumption reinforcement. that CRFBP is not only a sequestering protein, but it may possess additional functions. Neural correlates of an alcohol-cued Go/NoGo task: a dual process approach to binge drinking in Dysregulated endocannabinoid signaling in the college students central nucleus of the amygdala (CeA): role in Javier Blanco-Ramos (Santiago de Compostella, Spain) anxiety and excessive alcohol consumption Antonia M Serrano1 (1 Málaga, Spain) Dual-process models have been proposed as an explanation of substance abuse and other risky behaviours, with inefcient Te endogenous cannabinoid system is a neuromodulatory decision-making abilities as the central core. Decision making system that plays a homeostatic role in the constraint and abilities result from the balance between a refective system termination of stress responses. Disrupted endocannabinoid and an automatic or afective system. Te adolescent brain, signaling is linked to maladaptive stress responses and afec- in comparison with normal adults, presents an imbalance tive disorders such as anxiety and depression. Tese negative between earlier developed motivational systems and a still emotional symptoms are associated with alcohol dependence

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 243 Congress and abstinence and may contribute to relapse drinking Substance P (SP) and its preferred target – neurokinin 1 and excessive alcohol intake. We have shown that stress- (NK-1) receptors – have emerged as critical players in stress- and alcohol-induced increases in the endogenous levels of elicited alcohol seeking and alcohol consumption. However, 2-arachidonoylglycerol (2-AG) in the CeA were blunted in the underlying cellular mechanisms are still poorly under- rodents with a history of alcohol dependence. Tis efect was stood. SP and NK-1 receptors are widely expressed in the associated with anxiety-like behaviors and excessive alcohol brain and SP is released in response to stressful or painful consumption, and these dependence-associated behavioral stimuli. Here, we will discuss the efects of SP and a specifc efects were alleviated by enhancement of 2-AG tone. Tese NK-1 on GABAA receptor-mediated results suggest that dysregulated 2-AG signaling in the CeA neurotransmission in the CeA of diferent rodent models may contribute to dependence-related afective disorders and of alcohol dependence as well as innate anxiety. Briefy, SP excessive alcohol intake. increases transiently frequency and amplitudes of sponta- neous inhibitory postsynaptic currents (sIPSCs) in the CeA A role for amygdalar endocannabinoid signaling in of alcohol-naïve rats suggesting increased GABA release and excessive alcohol intake and comorbid anxiety in enhanced postsynaptic GABAA receptor-mediated neuronal genetically selected Marchigian Sardinian alcohol inhibition. Te NK-1 receptor antagonist decreases sIPSCs preferring rats frequency indicative of SP release into the CeA under basal 1 1 Nazzareno Cannella ( Camerino, Italy) conditions and a SP participation in regulating neuronal activity. Most notably, SP efects on CeA GABA transmis- Addiction is a chronic disease characterized by compul- sion are more pronounced and more sustained in alcohol- sive drug seeking and taking. Transition from recreational dependent rats. Tese functional efects are accompanied by drug use to excessive consumption and eventually drug decreased SP and NK-1 receptor expression pointing towards addiction is shaped by the interaction of excessive drug increased NK-1 receptor sensitivity. Similarly, larger and consumption with genetic, stress, and environmental factors. more sustained SP-induced increases of CeA GABAergic The Marchigian Sardinian alcohol-preferring (msP) rat transmission are observed in rats undergoing alcohol with- line, genetically selected for excessive alcohol consumption, drawal. Collectively, these data support the hypothesis that have a hyperactive amygdalar corticotropin releasing factor both alcohol dependence- and withdrawal-associated stress (CRF) to CRF receptor-1 signaling resulting in increased sensitize SP/NK-1 receptor signaling. Fatty Acid Amide Hydrolase (FAAH) activity and reduced N-arachidonoylethanolamine (AEA) levels, compared to non-selected Wistar controls. MsP rats show innate traits re- The endocannabinoid system and alcohol-related sembling generalized anxiety and post-traumatic stress disor- outcomes: genes, brain and behavior der (PTSD) in humans. Negative environmental conditions exacerbate this disorder, which can be attenuated by voluntary Christian S Hendershot (Toronto, Canada) alcohol drinking or by enhanced endocannabinoid transmis- sion. Here we provide evidences that FAAH inhibition in the central amygdala (CeA) decreases alcohol self-administration Endocannabinoid genes and alcohol-induced and stress-induced anxiety in msP but not in non-preferring reward phenotypes Wistar rats. We also compared alcohol intake and comorbid Vijay A Ramchandani1, Matthew E Sloan1, Emily L Vogt1, anxiety in male and female msP and Wistar rats demonstra- 1 1 1 ting that female msPs consume a larger amount of alcohol Melanie L Schwandt , Hui Sun , Peter Herscovitch , Markus 1 1 1 1 than male. Negligible alcohol consumption and no sex dife- Heilig , Nancy Diazgranados , David Goldman ( Bethesda, rences were observed in Wistars. Both male and female msP USA) show comorbid anxiety, however, while in male alcohol alle- viates generalized anxiety assed by an elevated plus maze pa- Background: previous studies indicate that endocannabinoid radigm, in female alcohol reduces the expression of a PTSD- (eCB) signaling may be related to the etiology of alcohol use like syndrome in a fear conditioning paradigm. In addition, disorder (AUD). Preclinical studies indicate that eCB admi- inhibition of FAAH activity mimic the efect of alcohol redu- nistered during alcohol intake increases mesolimbic dopa- cing PTSD-like behavior in female msP rats. Considering mine release. Cannabinoid receptor (CNR1) and fatty acid the link between reduced endocannabinoid transmission, amyl hydrolase (FAAH) gene variations have been associated excessive stress response, and expression of PTSD-like traits, with alcohol response and AUD severity. However, the broa- our data suggest that msP rats consume high amounts of der relationship between eCB and alcohol use has yet to be alcohol to normalize their amygdalar endocannabinoid trans- elucidated in humans. Aim: the aim of this study was to exa- mission in the attempt to medicate their innate negative state. mine the impact of endocannabinoid system polymorphisms CNR1 rs2023239 and FAAH rs324420 following IV alcohol Aberrant central amygdala Substance P/neurokinin infusion on striatal dopamine release measured using [11C]- receptor 1 signaling in rodent models of alcohol raclopride position emission topography (PET). Methods: dependence and anxiety twenty-six healthy males underwent two, randomized PET 1 1 Sophia Khom ( La Jolla, USA) scans with [11C]-raclopride, one concurrent with intravenous

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 244 Congress alcohol (target breath alcohol level of 80mg%) and the other Endocannbinoid-mediated effects of acute alcohol with placebo. The difference in [11C]-raclopride binding aministration in healthy humans 1 1 1 potential between alcohol and placebo sessions was used to Leah M Mayo , Elisabeth Paul , Robin Kämpe , Niclas quantify alcohol-induced dopamine release. Results: results Stensson1, Bijar Ghafouri1, Markus Heilig1 (1 Linköping, Sweden) show that CNR1 T/T homozygotes had signifcantly greater alcohol-induced dopamine release in the posterior (p=0.001) Background: chronic alcohol use is associated with dysre- and anterior ventral striatum (p=0.025) than C-allele car- gulation of the endocannabinoid system (eCB), a neuromo- riers. Tere was no impact of the FAAH polymorphism on dulatory system implicated in stress and reward processing. alcohol-induced dopamine release. To explore interactions However, little is known regarding the acute efects of alcohol between eCB and opioidergic mechanisms, a linear model on the eCB system in humans. Aims: our goal was to assess combining CNR1 and OPRM1 A118G genotype demons- the consequence of acute alcohol administration on circula- trated a signifcant additive efect of both genotypes on alco- ting eCBs and subsequent relationship to the and hol-induced dopamine release (R2=0.43). Conclusion: these rewarding efects of alcohol as assessed via functional magne- results suggest an additive efect of opioid and cannabinoid tic resonance imaging (fMRI). Methods: thirty-two healthy systems on striatal dopamine release following alcohol. Future adults (16 each men, women) participated in a within-sub- studies should explore the efect of eCB polymorphism on ject pharmacological fMRI study consisting of two sessions alcohol measures across the spectrum of AUD. (placebo, alcohol) on separate days. After drink consumption, they completed an fMRI scan assessing threat reactivity and A laboratory-based investigation of FAAH C385A reward processing. Blood samples were collected at baseline and alcohol-related phenotypes in youth and before, during, and after the scan to assess plasma eCB 1 1 1 Christian S Hendershot , Jeffrey D Wardell , Laura M Best , levels. Breath-alcohol concentrations (BrAC; target=0.06g%) 1 1 1 Rachel F Tyndale , Isabelle Boileau ( Toronto, Canada) and subjective mood and drug efects were assessed repeate- dly throughout sessions. In addition, participants were Background: the FAAH gene encodes the fatty acid amide genotyped at eCB-relevant loci (e.g. FAAH rs324420 and hydrolase (FAAH) enzyme, which metabolizes the endocan- CNR1 rs2023239). Results: alcohol signifcantly infuenced nabinoid anandamide. Te A allele of the FAAH C385A circulating eCBs, but this effect differed between sexes. variant (rs324420) has been linked to reduced enzyme acti- Specifcally, alcohol consumption increased eCBs in women, vity, higher anandamide, and increased severity of alcohol use but decreased eCB levels in men. Preliminary analyses indi- in adults. Aims: to extend this fnding to a younger sample, cate that the anxiolytic efects of alcohol (e.g. attenuation of we investigated associations of C385A with alcohol-related amygdala reactivity to threat cues) appear to be mediated, in phenotypes among heavy-drinking youth. Methods: partici- part, by eCB levels. Current analyses are underway to deter- pants (N=283, mean age=19.74 years) completed a self-report mine if eCBs similarly modulate reward processing during battery. A subset of participants completed alcohol adminis- alcohol intoxication. Conclusion: these data provide insight tration sessions involving the alcohol clamp (target BrAC= into acute biochemical consequences of alcohol that may 80mg%; n=88) and intravenous alcohol self-administration contribute to individual diferences in alcohol use and misuse. (IVASA) (n=61). Results: relative to youth with the C/C Are deficits in brain endocannabinoid metabolism genotype, those with the A allele (37%) reported greater alcohol consumption (AUDIT-C; p=0.047), hazardous use linked to heavier alcohol use? Neuroimaging (AUDIT; p=0.075), and stronger coping motives for drin- studies of the enzyme fatty acid amide hydrolase 1 king (p=.014). Coping motives mediated the association of Laura M Best , Bernard Le Foll, Esmaeil Mansouri, Richard genotype with consumption and alcohol-related problems. Bazinet, Dina Lagzdins, Pablo Rusjan, Rachel F Tyndale, During the alcohol clamp, within-subjects analyses of sub- Christian S Hendershot, Markus Heilig, Junchao Tong, Stephen jective responses revealed a signifcant, positive association J Kish, Isabelle Boileau (1 Toronto, Canada) between sedation and craving for A allele carriers (p=0.045), but not CC participants (p=0.138). Among CC participants, Background: fatty acid amide hydrolase (FAAH) is the within-person increases in sedation during IVASA predicted catabolic enzyme for the major endocannabinoid neurotrans- within-person decreases in craving, which in turn predicted mitter anandamide and a target for medication development. lower self-administration (indirect efect p=0.002). Tis efect Preclinical and genetic studies of a functional polymorphism was not observed for A allele carriers. Between-subjects in the FAAH gene (C385A, rs342240) suggest that lower analyses showed no genotype diferences on peak BrAC FAAH levels might be associated with risk for alcohol use during IVASA. Conclusion: fndings support an association disorder (AUD). Aim: to investigate whether lower brain of the C385A variant with self-reported consumption in FAAH level is associated with AUD, family history of AUD young drinkers. Results also implicate negative reinforce- and/or behavioural phenotypes related to risk for AUD. ment processes, including the motivational salience of acute Methods: FAAH brain levels were measured with positron efects, as candidate behavioral mechanisms for this emission tomography using the FAAH radioligand [11C] association. Funding: supported by the Canadian Institutes CURB in healthy controls (n=25), in heavy-drinking youth of Health Research, the Ontario Mental Health Research with positive (n=14) or negative family history of AUD Foundation, and NIH P60AA007611. (n=17) and in subjects with AUD at two time points (~5

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 245 Congress and ~25 days of monitored abstinence: n=14; n=11). Heavy- has hormonal efects, including increasing oestradiol levels, drinking youth completed an intravenous alcohol infusion which may afect the risk of breast cancer. Ethanol facilitates session and blood samples were taken in all participants to uptake of carcinogens in the mouth and throat, thus also assess FAAH C385A genotype and plasma endocannabinoid increasing the risk of tobacco-induced cancers. It may also levels at multiple time points. Results: [11C]CURB binding propel already existing cancers through immunosuppression, was globally lower than controls during early but not pro- angiogenesis, and decrease the efect of chemotherapeutics. tracted abstinence and signifcantly correlated with drinks per Alcohol use increases overall cancer incidence and overall and week and with plasma concentrations of anandamide. Family cancer-specifc mortality. Epidemiological and experimental history of AUD did not afect [11C]CURB binding, however research on alcohol and cancer remains limited. Research higher alcohol consumption and hazardous use (Alcohol Use priorities include 1) investigation of the role of alcohol in a Disorders Identifcation Test (AUDIT)) in heavy-drinking wider list of cancers; 2) better analytical strategies to elucidate youth, as well as lower sedative efects of alcohol during intra- the role of drinking patterns and of specifc alcoholic drinks; venous administration (Biphasic Efects of Alcohol Scale) and 3) elucidation of the role of smoking in alcohol-related was related to lower [11C]CURB binding. Conclusion: in carcinogenesis, particularly in those cancer sites that are line with preclinical studies our fndings that lower FAAH is tobacco-related. related to higher alcohol consumption (in AUD and in non- Role of adaptive immune system, particularly IgA+ AUD heavy drinking youth) may be an acute consequence and CD8+ T cells, in alcoholic steatohepatitis and of recent chronic alcohol use (in AUD) and or a predating its progression to hepatocellular carcinoma factor increasing vulnerability for hazardous use. Although 1 1 clinical signifcance of low FAAH in AUD remains to be Shabnam Shalapour ( La Jolla, USA) established, treatment approaches targeting FAAH should consider that increased endocannabinoid tone during early Hepatocellular carcinoma (HCC) is one of the most preva- abstinence could drive drinking however some aspects could lent malignancies worldwide and a leading cause of cancer- be benefcial. Funding: supported by the Canadian Institutes related deaths. Despite major growth in fghting hepatitis of Health Research, the Ontario Mental Health Research virus B and C, the epidemic of liver disease continues to Foundation, and NIAAA 1R21AA022246-01A1. grow with clear links to obesity and alcohol abuse. Non- alcoholic and alcoholic steatohepatitis (NASH/ASH) are the major drivers of HCC, but the mechanisms underlying Insights into alcohol-associated cancers the progression to HCC are poorly known. Progress in this feld depends on the availability of reliable preclinical models Sebastian Mueller (Heidelberg, Germany) amenable to genetic and functional analyses and exhibiting robust NASH-to-HCC progression. Using MUP-uPA mouse models, we found that accumulation of IgA+PD- Alcohol and cancer: the epidemiological evidence L1+CD138+IL-10+ plasmocytes in NASH-aficted human 1 1 1 1 Elisabete Weiderpass , Carolina Espina , Pietro Ferrari ( Lyon, and mouse livers results in localized immunosuppression that France) fosters HCC development by attenuating the activation of a protective, tumor-directed cytotoxic CD8+ T-cell response. Alcohol consumption is one of the top-10 health risks, Moreover, we found that alcohol feeding increases tumor contributing to circa 3.9% of worldwide burden of disease development through a profound induction of gut barrier and 3.3 million annual deaths. Alcohol use increases overall disruption, and systematic changes in adaptive immune cells’ cancer incidence and overall and cancer-specifc mortality. development and responses. Although high fat diet (HFD) Te 2012 IARC Monograph reviewed the epidemiological supports CD8+ T cell infltration in the liver, alcohol sup- evidence on the possible association between alcoholic beve- presses it. Our work provides new insights on how HFD rage consumption and cancer risk at 27 anatomical sites, and and alcohol regulate adaptive immune cells and thereby afect reported that cancers of the upper digestive tract (UADT; fbrosis and the response to immunotherapy. Specifcally, oral cavity, pharynx, larynx, esophagus), liver, colorectum and consumption of alcohol or HFD regulates the response to female breast are causally related to the consumption of alco- anti-PD(L)1 therapy in a diferent manner, namely due to holic beverages. In 2018 the World Cancer Research Fund/ its distinct ability to regulate CTL function and induction of American Institute for Cancer Research (WCRF/AICR) dysbiosis, both locally and systemically. report additionally showed that alcohol intake was inversely Molecular mechanisms of alcohol-associated associated with the risk of kidney cancer and described a sug- gestive positive relationship with pancreatic cancer. Tere are cancers: from metabolism and oxidative stress to several mechanisms through which alcohol induces cancer: stem cells and genomic instability 1 2 a direct efect on the cells where conversion to acetaldehyde Vasilis Vasiliou , David C Thompson (1 New Haven, USA, such as primarily in the liver cells, where it can induce cirrho- 2 Aurora, USA) sis; conversion also occurs in the saliva and the large intestine. Ethanol promotes production of highly reactive oxygen spe- Chronic ethanol abuse is associated with, and may be cies, which can damage DNA, alter DNA methylation, and involved with the development of, several types of cancers

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 246 Congress in humans, including liver, colon, pancreas and, in women, the aldehyde dehydrogenase 2 gene, which has randomized breast. Although the exact cause of alcohol-induced cancer is millions of alcohol consumers to markedly increased ACH currently unknown, several possible underlying mechanisms exposure via saliva and gastric juice. Tis novel human cancer have been suggested and are currently under investigation. model is associated with a manifold risk for upper GI tract It is well established that alcohol metabolism promotes cancer and proves conclusively the causal role of local ACH oxidative stress. Of the many molecules formed during in alcohol-related upper digestive tract carcinogenesis. Most ethanol metabolism, the aldehydes exhibit activities that importantly, the model minimizes the role of confounding can promote cancer. Acetaldehyde and lipid peroxidation factors hampering most epidemiological studies on alcohol aldehydes (generated by reactive oxygen species during and cancer. Normal human saliva does not contain mea- CYP2E1-mediated alcohol metabolism) can form DNA surable levels of ACH. However, alcohol ingestion results adducts, and thereby cause genetic mutations, inhibit DNA within seconds in a concentration-dependent accumulation repair, and disrupt DNA replication processes, all of which of ACH in saliva, which continues up to 10-15 minutes after cause genomic instability – a hallmark of cancer. In addition, each sip of alcoholic beverage. Te prominent instant increase aldehydes may adduct proteins and inhibit cellular processes, of salivary ACH level is followed by a long-term phase las- such as folate and retinoid metabolism. Tey can also regulate ting for as long as ethanol stays in the saliva. Bacteria and redox-sensitive signaling pathways and transcription factors yeasts representing normal upper GI tract microbiome play that sustain infammation. Alcohol consumption afects the a major role in local ACH formation from ethanol. Tis is composition and function of the gut microbiome, leading to contributed locally by organ specifc expression and gene infammation and changes in the function of the immune polymorphisms of ethanol- and ACH-metabolizing enzymes. system, both of which are known to promote and exacerbate References cancer. Importantly, gastrointestinal bacteria can produce - Lachenmeier DW, Salaspuro M, Regul Toxicol Pharmacol 2017;86:128-136. acetaldehyde from ethanol. Finally, reactive oxygen species - Nieminen MT, Salaspuro M, Cancers 2018;10;11; doi:10.3390/can- cers10010011 and lipid aldehydes may also afect immunometabolism that is now recognized as an important factor in cancer. As such, there are many mechanisms by which aldehydes can contri- Novel concepts in the evaluation of fibrosis in bute to alcohol-associated cancers. As the primary enzymes responsible for detoxifying aldehydes, aldehyde dehydro- alcohol-related liver disease genases (ALDHs) have the potential to infuence alcohol- associated carcinogenesis. Indeed, we and others have shown Carolin Lackner (Graz, Austria) ALDHs to be involved in the pathophysiology of cancer and cancer stem cells. We now propose two novel hypotheses Non-invasive fibrosis assessment in compensated regarding the role of ALDHs in alcohol-mediated carci- and decompensated alcohol-related liver disease nogenesis. First, ALDHs may shield tumor-initiating or tumor cells against genomic instability by protecting their and clinical implications 1 1 DNA repair mechanisms from damage by acetaldehyde and R. Stauber ( Graz, Austria) lipid-derived aldehydes. Second, ALDH-derived acetate could serve as a precursor of acetyl-CoA, a fuel used to form Alcohol-related liver disease (ALD) is the most common biomolecules in cancer cells. In conclusion, the mechanisms liver disease in the Western World. Especially in Europe by which alcohol metabolites and associated oxidative stress ALD is a substantial burden as the European population may contribute to carcinogenesis are complex and often has the highest per capita alcohol consumption world-wide. interconnected. Teir elucidation provides new opportunities However, awareness about the risks of heavy drinking is low for the prevention or mitigation of alcohol-associated cancers. among patients with ALD and they usually present at an advanced stage with clinical symptoms such as jaundice and/ local acetaldehyde – its key role in alcohol-related or ascites. Early/compensated ALD does not manifest itself upper gi tract carcinogenesis clinically and patients rarely seek medical advice, unless ALD Mikko Salaspuro1 (1 Helsinki, Finland) is detected during work-up of other illnesses. Screening for early ALD is warranted since it is readily reversible under Ethanol molecule is neither genotoxic nor mutagenic. abstinence and treatment is available (psychological interven- However, its frst metabolite acetaldehyde (ACH) associated tions, anti-craving drugs). Recent data show that prognosis with the consumption of alcoholic beverages is classifed as of early ALD, apart from abstinence, is mainly determined carcinogenic in humans. ACH concentrations present in by histological fbrosis stage; advanced fbrosis (F3, bridging saliva instantly after alcohol drinking result in the generation fbrosis; F4, cirrhosis) carries dismal outcome (10-year mor- of mutagenic ACH-DNA adducts in human mouthwash tality 45%). However, due to the high prevalence of ALD, samples and in oral mucosa of rhesus monkeys. Recently, universal liver biopsy is not feasible. Noninvasive fbrosis tests similar ACH levels have been shown to play a key role in have shown high diagnostic accuracy for advanced fbrosis ethanol-dependent telomere shortening in primary human in a variety of liver diseases including ALD. Among these, foreskin fibroblasts. Strongest evidence for the local carci- simple fbrosis tests such as FIB-4 are based on routine cli- nogenicity of ACH in man provides a point mutation in nical and laboratory parameters and provide relatively high

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 247 Congress diagnostic accuracy at low cost. Proprietary fbrosis panels of cirrhotic fbrosis with diverse etiology, we developed a based on direct fbrosis markers including Enhanced Liver dual-photon microscopy-based computerized image analy- Fibrosis (ELF™) test, FibroMeter™ and FibroTest show sis tool to detect architectural features including improved diagnostic accuracy. Vibration-controlled transient both collagen geometry and collagen spatial network within elastography (VCTE, FibroScan®) enables noninvasive esti- septa, nodule, and sinusoidal regions of interest. A profle of mation of liver stifness (LS) and has shown superior dia- these collagen features composed the quantitative collagen gnostic accuracy in comparative studies. However, LS is not pattern (QCP) of the detected biopsy specimen. By using only determined by fbrosis but also infammation, cholestasis this tool, we analyzed a retrospective cohort, which included and/or hepatic congestion. Importantly, several studies have 225 hospital patients diagnosed with acute decompensation demonstrated a rapid decline of LS during alcohol detoxifca- of biopsy-proven cirrhosis, with clinical collaboration from tion, which presumably refects the resolution of steatohepati- the Institute of Liver and Biliary Science, New Delhi, India. tis. A recent comparative study showed best performance for We found that QCP faithfully captured the morphologic VCTE (per protocol) and ELF score. While the latter tests diference in fbrosis patterns between etiologies of fatty liver are mostly restricted to referral centers, simple tests such as diseases and viral hepatitis and between Laennec stages. It FIB-4 are suitable for screening in primary care. Te clinical also related to MELD and HVPG. When using for pre- impact of noninvasive fbrosis tests in decompensated ALD is dicting the short-term mortality in these patients, QCP rather limited since most patients already have clinical signs had a robust prognostic performance superior to MELD, of cirrhosis. CTP, Laennec stage and collagen proportionate area. On multivariate regression, it was identifed as an independent Staging of alcohol-related liver disease and clinical prognostic factor. All the data indicate that histologic fbrosis implications captured by QCP could have an important prognostic value 1 1 C. Lackner ( Graz, Austria) even in the end stage of liver disease. In addition, because by using the same measurement strategy, we previously found Histological stage has emerged as one of the most important that cirrhosis remodeling in viral hepatitis can be detected in predictors of outcome in most chronic liver diseases including high accuracy, we would propose that QCP may be helpful non-alcoholic fatty liver disease (NAFLD), chronic viral for assisting the investigation of undiscovered meaning of hepatitis and autoimmune liver diseases. Surprisingly, despite histologic fbrosis with diverse chronic liver disease. the fact that ALD is among the most frequent liver diseases, to date no universally accepted histological staging system has been devised. Because NAFLD and ALD share broad Ethanol-induced neuroinflammation and morphological overlap, also with respect to type of fbrosis oxidative stress: chronic ethanol intake and and its centrilobular predominance, several authors have pro- binge-like intoxication – translational options posed that histological staging systems for NAFLD may also be used for ALD. However, there are points of concern with such an approach. Firstly, most patients with compensated Yedy Israel (Santiago, Chile) as well as decompensated ALD are cirrhotic at frst presen- tation. Tere are diferent histological stages of severity of It is well accepted that chronic alcohol intake or its admi- cirrhosis which are not refected in most staging systems used nistration lead to neuroinfammation; seen as morphological in NAFLD. Secondly, there are data suggesting that pericel- changes in astrocytes and microglia. Tese are clearly seen lular and septal fbrosis types may have diferent impact on in alcohol-consuming rodents, while microglial changes outcome in patients with ALD. An ALD-specifc staging in humans presenting alcohol-use-disorders (AUD) may system has recently been devised by the Consortium for depend on the noninvasive detection method employed. the Study of Alcoholic LiVer Disease in Europe (SALVE) Neuroinfammation and oxidative stress are self-perpetuated Histopathology Group. Tis ALD-specifc staging system for prolonged periods in a vicious-like cycle and appear to be (SALVE stage) refects diferent histological stages of severity the basis for protracted increases in chronic ethanol intake of cirrhosis and the degree of pericellular fbrosis in ALD. and relapse-like binge drinking. Te presentations in this Te potential clinical implications of SALVE staging will Symposium show (i) clear microglial changes, as determined be discussed. by PET imaging in AUD patients; (ii) demonstrate marked reductions in ethanol intake induced by antioxidant N-acetyl Histologic fibrosis patterns associate with disease cysteine both in operant and chronic ethanol administration severity and short-term mortality in patients with paradigms in rodents, including the abolition of binge-like acute decompensation of cirrhosis drinking in the post-deprivation and re-access condition Yan Wang1 (1 Guangzhou, China) (ADE) (iii) show that N-acetyl cysteine normalizes the both oxidative stress and the glial changes induced by ethanol, in Histologic fbrosis has an important prognostic value in line with the vicious cycle hypothesis (iv) demonstrate that natural history of chronic liver disease. However, there are the systemic or intranasal administration of mesenchymal few evidences for its role in the end stage of liver disease. stem cells (MSC) or MSC-products, presenting marked anti- For the purpose of fully describing the morphologic features infammatory and antioxidant properties, strongly inhibit

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 248 Congress ethanol-induced and relapse drinking. Tese studies further administration, alcohol-seeking behavior and to reduce show that MSCs normalize the ethanol-reduced levels of the relapse on rats that were abstinent for 17 days. In a “post- glial glutamate transporter GLT-1. Overall the Symposium dependent” state rat model (induced by chronic intermittent ties neuroinflammation/oxidative-stress/hyper-glutama- ethanol (CIE) vapour exposure for 10 weeks in male Wistar tergic conditions as the likely mechanism that perpetuates rats), we evaluated the efects of NAC during acute with- chronic alcohol intake and promotes intoxicating relapse. drawal, 8 hours after inhalation chambers were turned of. We Translational options are envisioned. showed that a lower dose of NAC (25 mg/kg) was enough to reduce ethanol self-administration and motivation to TSPO PET imaging to study the dynamics of consume ethanol, evaluated in a progressive ratio paradigm, ethanol-induced neuroinflammation while the 50 mg/kg NAC reduced extinction responding and 1 1 Wadad Saba ( Orsay, France) reacquisition of self-administration after 1 month abstinence. Overall, our results demonstrate that NAC is able to limit Imaging techniques play an important role for the non-in- rat’s ethanol self-administration, extinction responding, and vasive determination of the efects of alcohol in vivo, mainly relapse, making it a potential new treatment for the mainte- focusing on brain structure and neuronal function. Positron nance of abstinence via an anti-craving efect. We will discuss Emission Tomography (PET) imaging using radioligands further the possibility to evaluate NAC potential as a new of the translocator protein 18 kDa (TSPO), a biomarker of treatment of AUD in patients. glial activation, is useful to address the importance of neuro- infammation in various pathophysiological states and ofers Administration of anti-inflammatory mesenchymal a unique tool to study the dynamics of the neuroimmune stem cells or its secretome inhibits alcohol impact of alcohol exposure on the brain. In adolescent mon- self-administration and blocks relapse intake. keys, TSPO PET imaging using 18F-DPA-714 revealed that Mechanisms and translational opportunities an acute and initial ethanol exposure (0.7-1.0 g/L) induced Fernando Ezquer1, Maria Elena Quintanilla1, Paola Morales1, an immediate and prolonged (7-12 months) glial activation, Daniela Santapau1, Pablo Berrios-Cárcamo1, Marcelo Ezquer1, suggesting a priming of glial function after initial alcohol Mario Herrera-Marschitz1, Yedy Israel1 (1 Santiago, Chile) exposure. In rats chronic alcohol exposure over a 14-day period induced an increase in the binding of 18F-DPA-714 Chronic alcohol consumption leads to neuroinfammation which was reduced by (0.4 mg/kg, s.c, 1 hour prior and brain oxidative stress, which inhibit the astrocyte Na- to ethanol injection). Tis unexpected efect could be linked glutamate transporter (GLT-1), proposed to perpetuate to the antagonist property of nalmefene on Toll-like receptor alcohol intake and relapse. Mesenchymal stem cells have 4 (TLR4). Our results are consistent with neuroinfamma- been postulated as a therapeutic option for the treatment of tion associated with acute/chronic alcohol exposure. Tis diferent diseases since they can produce potent anti-infam- contrasts with the decreased binding of 11C-PBR28, another matory molecules and reduce oxidative stress. Studies pre- TSPO ligands, observed in the brain in patients with alcohol sented evaluate the addictive-like suppression exerted by (i) use disorders, which is consistent with a blunted peripheral intracerebroventricular administration of anti-infammatory proinfammatory response compared with controls. TSPO mesenchymal stem cells (MSCs), (ii) intravenous adminis- PET imaging provides a novel insight into the dynamics of tration of MSC-spheroids or (iii) intranasal administration glial function related to alcohol exposure and may be useful of MSC-derived secretome in a rat model of chronic ethanol to i) address the neuroimmune component of alcohol-related intake and relapse drinking. Studies show that administra- neurotoxicity and addiction and ii) evaluate immunothera- tion of a single intracerebroventricular or intravenous dose peutic strategies for or the treatement of of MSCs or the administration of three intranasal doses of alcohol dependence. MSC-derived secretome: (a) inhibited chronic ethanol intake and relapse binge drinking by 80-90%, displaying protrac- Evaluation of N-acetylcysteine effects on two ted efects over 4-5 weeks; (b) fully normalized alcohol- preclinical rat model of Alcohol Use Disorders: induced neuroinfammation, shown as a reduced astrocyte the operant binge drinking model and the “post- and microglia activation in hippocampus; (c) reduced brain dependent” state rat model oxidative stress evidenced by the restoration of the normal 1 1 Catherine Vilpoux , Sophie Lebourgeois , María Carmen hippocampal GSH/GSSG ratio and (d) markedly increased González-Marín1, Mickael Naassila1 (1 Amiens, France) the levels of the GLT-1 transporter in prefrontal cortex and nucleus accumbens. Knockdown of GLT-1 transporter by Many components of ethanol addiction such as reinforce- administration of an antisense oligonucleotide fully abolished ment, withdrawal, extinction, and relapse are known to in- the inhibitory efect of MSC-derived secretome on ethanol volve glutamate transmission, and N-acetylcysteine (NAC) is intake, suggesting that the glutamate transporter GLT-1 thought to counteract glutamatergic dysregulation underlying mediates the addictive-like MSCs inhibitory efects. Overall, ethanol addiction. We tested NAC efect on two diferent rat studies indicate that the administration of anti-infammatory models of Alcohol Use Disorders (AUD). In a rat model of mesenchymal stem cells or its secretome afords translational operant binge drinking, we demonstrated the efcacy of acute opportunities for the treatment of alcohol-use disorders. 100 m/kg NAC treatment to reduce ethanol alcohol self- Supported by FONDECYT #1180042 to YI.

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Translational studies on ethanol effects in adult hippocampal neurogenesis, reduced forebrain choliner- adolescence gic and midbrain serotonergic neurons as well as increases in markers of neurodegeneration. Post-mortem AUD human Cindy L Ehlers (La Jolla, USA) brain also indicated increases in HMGB1, toll-like receptors, RAGE, interferon, CCL2 and other signaling genes. Further, increases in human AUD brain gene expression correlated Neurophysiological synchrony and sleep are both with age of drinking onset and lifetime alcohol consumption. disrupted in young adult humans and rats with a Emerging studies fnd exercise, indomethacin, and history of adolescent alcohol exposure galantamine prevent and/or reverse AIE pathology Tese fn- Cindy L Ehlers1 (1 La Jolla, USA) dings support the hypothesis that adolescent alcohol exposure increases expression of HMGB1, TLR, RAGE, NFkB, and The present study aimed to document the young adult other genes are modifed through epigenetic gene silencing consequences of adolescent alcohol exposure, in humans and or enhancing signals that persist into adulthood long after rodents, as assessed by waking EEG and sleep. Synchrony alcohol exposure ends, but are in some cases are reversible. of phase (phase-locking, PL) of event-related oscillations Tese fndings identify targets for consideration of treatment (EROs) between frontal and parietal cortex and sleep data, of adolescent onset AUD. Supported by NIAAA AA020024, were evaluated. Te human participants were young adults AA020023, AA011605. (age 18-30 yrs, n=1041), with and without a history of adolescent binge drinking (5 drinks for boys 4 for girls per Impulsive action and decision making in young occasion at least once per month), and 74 young adult rats adults binge drinkers; brain mechanisms with and without a history of 5 weeks of adolescent alcohol Theodora Duka1 (1 Brighton, UK) vapor exposure (PD 23-55). Human binge drinkers were found to have lower PL in the beta and theta frequencies Binge drinking is associated with increased impulsivity. between frontal and parietal cortex. PL was also decreased Data will be presented to elaborate on the role of impul- in the rats exposed to ethanol vapor in the theta band across sivity facets and brain function in alcohol abuse. In young the two cortical regions. A history of adolescent regular adult binge drinkers (BD; aged 18 to 25) the relationship binge drinking was also associated with reduced sleep qua- between “motor” impulsivity in the form of “can’t stop“ and lity as indexed by: longer sleep latencies, more problems “can’t wait” as well as “temporal” impulsivity (failure to delay with breathing, bad dreams and an overall higher Pittsburgh gratifcation) is examined. In parallel, “can’t wait” impulsivity sleep quality index total score. Adolescent vapor exposure in is tested in two inbred strains of mice known to difer in the rat was found to result in decreases in theta power (4-8 alcohol intake (alcohol preferring and alcohol averse mice). In Hz) and delta (1-4 Hz) and theta (4-8 Hz) power during addition functional brain activity and resting state functional slow wave sleep (all p’<0.05). Tese fndings suggest that connectivity is tested in BD. Binge drinkers showed robust alcohol exposure during adolescence may result in defcits in impairments in “can’t wait” impulsivity under increased atten- sleep quality in humans and slow wave sleep in animals and tional load; alcohol preferring mice also showed impairments decreases in synchrony between cortical neuronal networks, in “can’t wait” impulsivity compared to alcohol-averse mice in both species. Supported by R37 AA010201, AA026248, before any exposure to alcohol. Brain imaging revealed that RO1 AA027316, U01 AA019969. higher BD severity is associated with enhanced activation in Adolescent alcohol exposure alters adult precentral gyrus and superior parietal lobule during success- neurobiology through neuroimmune and ful stop responses, indicating a compensatory mechanisms. epigenetic signaling Delayed gratifcation was associated with lower frontopolar Fulton Crews1 (1 Chapel Hill, USA) activation. Resting-state functional connectivity revealed that the higher the incidence of BD, the lower the coupling of the right supramarginal gyrus to the Ventral Attentional Adolescence brain develops in parallel with maturation of Network (VAN). Tese fndings support the assumption that self-control. To investigate the impact of adolescent binge aspects of cognitive impairments seen in binge drinkers in drinking on brain development Wistar rats were exposed particular those associated with “can’t wait” impulsivity may to adolescent intermittent ethanol (AIE, 5gm/kg/day-2 precede drinking behaviour. Disrupted functional connecti- days on-2 of ) across puberty and assessed in adulthood vity within the Ventral Attention Network in more binging for cognitive defcits and changes in neurobiology. Post- individuals may suggest disrupted attentional processing pro- mortem human brain of controls or AUD patients were also viding supporting evidence for the brain signature associated determined. AIE increased adult brain expression of innate with binge drinking. immune genes HMGB1, CCL2, Toll-like receptors, and RAGE. AIE also increased activation of the transcription Epigenetic, neuroinflammation and GluN2B factor NFkB and altered glial morphology in parallel with participate to cognitive deficits after ethanol increases in histone methylation, H3K9me2, a marker of binging in adolescent rat epigenetic silencing. AIE also increased adult risky decisions 1 1 and blunted behavioral fexibility in parallel with reduced Olivier Pierrefiche ( Amiens, France)

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Binge drinking induces memory impairment and recently, we the family Anaeroplasmataceae to be more abundant. No showed that only Two Ethanol Binge-like Exposure (TEBE) significant difference was found between Conditions A in adolescent rats are sufcient to transiently abolish long- and B. Our fndings suggest that in alcohol binge drinking term synaptic depression (LTD) in the hippocampus leading baboons, long-term exposure to alcohol binge drinking (G3) to mnesic defcits after 48h. To understand the mechanism of leads to signifcant changes in the gut microbiome, whereas such long-lasting action of EtOH, we investigated the role of short-term (G2) does not. Tese changes were not afected epigenetic and neuroinfammation after TEBE in the hippo- by acute short-term forced abstinence. Tis was confrmed in campus of young adult rats. Neuroinfammation was revealed a rat model of binge alcohol drinking, where we see similar through an increase in TLR4 immunolabelling in CA1 area reduction in microbiome diversity after prolonged exposure, and in vimentin + GFAP co-labelling showing astrogliosis in and this whether the rats were knock-out or not for the the dentate gyrus (DG). Neurogenesis was revealed with an receptor of the feeding-related hormone ghrelin. Our result increase in doublecortin labelling in the subgranular zone of support a role for the gut microbiome in binge drinking. the DG. In stratum oriens of CA1, synaptic pruning probably Implication of the gut microbiota in the occurs since synaptophysin labelling decreased. Expression behavioral changes linked to alcohol-dependence: level and activity of Histone Deacetylase 2 (HDAC2), mechanistic approach involved in epigenetic, increased while acetylated Histone 4 1 1 1 (Ac-H4) decreased. Further, TEBE increases mRNA level Sophie Leclercq , Tiphaine Le Roy , Laure Bindels , Caroline 1 1 1 1 for GluN2B subunit of the NMDA receptor while ChIP Quoilin , Audrey Neyrinck , Peter Stärkel , Philippe de Timary , 1 1 analysis revealed that HDAC2 modulates the GluN2B gene Nathalie M Delzenne ( Brussels, Belgium) promoter. Further, TEBE altered GluN2A/GluN2B balance in synaptic transmission. Finally all cellular efects of TEBE It is well established that alteration of the gut microbiota were prevented with sodium butyrate, an HDAC inhibitor. composition can disturb many aspects of host physiology, In conclusion, two EtOH exposures induces long-lasting including metabolism, immunity and peripheral and central memory-impairment because it overexpressed HDAC2 nervous system with consequences for brain functions and resulting in GluN2A/GluN2B imbalance that leads to LTD behavior. In a previous study, we showed that alterations blockade. In parallel, neuronal injury and altered morphologic of the gut microbiota composition of alcohol-dependent plasticity in the hippocampus take place. It is now important (AD) patients were associated with high score of depression, to study the link between infammation and epigenetic in anxiety and alcohol craving, suggesting the existence of a gut- the efects of ethanol since this would help developing new brain axis in AD patients. Here, we demonstrated the causal therapeutic strategy. role of the gut microbiota in the development of the psy- chological symptoms associated with alcohol dependence, by using fecal microbiota transplantation. Te microbiota of AD New arguments for a role of the gut microbiota patients and healthy controls (CT) were transferred into two and inflammation in alcohol-use-disorders groups of mice which were subsequently tested for behavior. We found that mice transplanted with the gut microbiota Philippe de Timary (Brussels, Belgium) of AD patients exhibited increased depression-like behavior and decreased social behavior compared to CT-recipient mice. Furthermore, AD-recipient mice showed increased The gut microbiome in binge alcohol drinking: infammatory and activated microglia markers in recent translational efforts the striatum, decreased expression of myelin-related genes 1 1 Lorenzo Leggio ( Providence, USA) in the frontal cortex and unbalance of GABA/glutamate neurotransmission. Metabolomics analysis revealed that a We provided the frst descriptive analysis of the gut micro- specifc metabolite might be responsible for these changes biome in a unique non-human primate model of alco- in brain functions and behavior observed in AD-recipient hol binge-drinking. We analyzed the gut microbiome on mice. Tese results strongly reinforce the existence of gut- fecal samples from male baboons chronically exposed to brain interactions in mental disorders, and highlight the gut either alcohol or a non-alcoholic isocaloric beverage (tang). microbiota as a new potential target in the management of There were three treatment groups: G1=tang (controls); alcohol addiction. G2=“short-term” alcohol binge drink (2-3 years); G3=“long- The gut microbiota as a new target in the term” alcohol binge drink (10 years). Fecal samples were collected in two conditions: A=early abstinence (days 3-5) treatment of disinhibition in alcohol-dependence: and B=during 3 days of ongoing drinking. Microbial alpha- a clinical study diversity was signifcantly lower in the G3 group vs. the G1/ Caroline Quoilin1, Julie Duque1, Philippe de Timary1, Sophie G2 groups. Te two genera Lactobacillus and Streptococcus Leclercq1 (1 Brussels, Belgium) showed high relative abundances in G3. Fecalibacterium was reduced in G3 only. For G2, the order Clostridiales and the Alcohol dependence is usually seen as a disinhibitory disor- family Ruminococcaceae showed high relative abundances der, notably characterized by a state of central nervous system compared to G1 and G3. Cohort G1 showed members of hyperexcitability and a lack of inhibitory control. In parti-

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 251 Congress cular, by applying transcranial magnetic stimulation (TMS) to plasma infammatory markers, to evaluate whether these over primary motor cortex (M1) to assess the excitability of changes could express brain infammation and to changes in the motor corticospinal pathway during action preparation, functional connectivity and behavior. we have shown that alcohol-dependent (AD) patients sufer from a defcit in physiological motor inhibition when plan- ning a behaviour. Recently, among the potential mechanisms CIFASD advances in the pathophysiology and underlying this inhibitory defcit, an alteration of the gut diagnosis of FASD microbiota composition has been identifed as a promising candidate. Here, we aimed at determining whether a treat- Michael E Charness (Boston, USA) ment with prebiotics, by restoring the microbiota, modifes motor cortex excitability and inhibitory abilities of AD pa- tients. To do so, 50 AD patients participated in a randomized, Molecular mechanisms underlying ethanol double blind, placebo-controlled clinical trial, in which they teratogenesis and its antagonism received a supplementation with dietary fbers (inulin) or X. Dou, J.Y. Lee, Michael E Charness1 (1 Boston, USA) a placebo during a 3-week detoxifcation program. Motor cortex excitability was examined at the end of the treatment, Ethanol teratogenesis is caused in part by ethanol disrup- using a range of TMS measures, including the resting motor tion of the L1 neural cell adhesion molecule (L1). Ethanol threshold, recruitment curve, short and long intracortical inhibits L1 adhesion by interacting with a binding site inhibition and intracortical facilitation within M1. Moreover, in the extracellular domain of L1 at the Ig1-Ig4 inter- physiological motor inhibition was assessed during action face. NAPVSIPQ (NAP) an octapeptide, blocks ethanol preparation by applying TMS in a choice reaction time task. inhibition of L1 adhesion and prevents ethanol teratogene- Finally, all patients completed questionnaires and performed sis in mouse embryos at femtomolar concentrations by an neuropsychological tasks to evaluate their level of impulsivity unknown mechanism. Ethanol inhibition of L1 adhesion and behavioral response inhibition. Tis double-blinded cli- requires L1 binding to ankyrin-G and the spectrin/actin nical study will allow to elucidate whether the gut microbiota cytoskeleton. Dephosphorylation of selected residues on is an efective target in the treatment of a core symptom of the L1 cytoplasmic domain (L1-CD) leads to uncoupling alcohol dependence. of L1 from ankyrin-G, rendering L1 insensitive to ethanol. Polymorphisms in the genes that encode p90rsk, a kinase that Does inflammation lead to changes in brain phosphorylates the L1-CD, and ankyrin-G were associated anatomy in alcohol use disorder (AUD)? The with facial dysmorphology in children with heavy prena- effects of alcohol withdrawal in MRI scans tal alcohol exposure. Ankyrin-G binding to L1 requires 1 1 1 Philippe de Timary , Géraldine Petit , Laurence Dricot , Sophie dephosphorylation of tyrosine in the FIGQY1229 ankyrin- 1 1 1 Leclercq , Pierre Maurage ( Brussels, Belgium) binding motif of the L1-CD. Femtomolar concentrations of NAP activated the phosphorylation of L1-Y1229 and the AUD is characterized by large brain morphological altera- dissociation of L1 and ankyrin-G. NAP phosphorylation tions. Alcohol-withdrawal, is attended by large behavioral of L1-Y1229 was mediated by EphB2, and knockdown of and infammatory changes that have been related to altered EphB2 abolished ethanol inhibition of L1 adhesion. Tus, microbiota composition. Here we tested by MRI scans NAP antagonizes ethanol inhibition of L1 adhesion by the hypothesis of a drinking related edema that would activating the phosphorylation of L1-Y1229, inducing the resolve during withdrawal. 19 AUD inpatients undergoing dissociation of L1 and ankyrin-G, and stabilizing an ethanol- a detoxifcation, were tested on the frst and 18th day of insensitive conformation of L1. Supported by NIAAA Grant withdrawal, for MRI brain anatomy and DTI. Results: using R01AA012974; U24AA014811 (CIFASD); VA Merit paired T tests, alcohol withdrawal was attended by signi- Review 5I01BX002374; and DoD W81XWH-12-2-0048 fcant decreases in volume of 4th ventricle, choroid plexus, Subaward 8742sc. white matter, while the cortical volume was increasing. In parallel to these changes, we also observed a decrease in Synergistic gene-environment interactions in mean difusivity in all the white matter regions tested, and a zebrafish model of Fetal Alcohol Spectrum these decreases were signifcant in 21 out of the 36 regions Disorders tested. A decrease in mean difusivity was also observed in J.K. Eberhart1, S. Tucker, Y. Fernandes1, N. McCarthy1 (1 Austin, grey matter in 160 of the 180 regions tested and signifcant USA) in 50 of these regions, but globally signifcant changes were observed in regions corresponding to the central executive, Susceptibility to FASD is genetically modulated, but the motor, salience, auditory, and most signifcant in the visual nature of these modifying loci is poorly understood. We and default-mode grey matter regions. In only one region have used genetic screens to identify mutants that are sensi- we observed a signifcant increase in mean difusivity : the tive to normally sub-teratogenic doses of ethanol. Embryos right pallidum. Conclusion: alcohol withdrawal is related to mutant or heterozygous for platelet-derived growth factor changes in brain anatomy and mean difusivity in both white receptor alpha (pdgfra) are exquisitely sensitive to ethanol- and grey matter regions. Tese changes will be compared induced facial defects. Under control conditions, proper

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1 1 2 3 neural crest cell migration, but not survival, requires Pdgfra Michael Suttie , Peter Hammond , Neil Aiton , CIFASD 1 2 3 function. Following ethanol exposure, Pdgfra promotes the ( Oxford, UK, Brighton, UK, Collaborative Initiative on Fetal survival of neural crest cells. Te PI3K pathway is the major Alcohol Spectrum Disorders, USA) Pdgfra efector, and the PI3K/mTORC1 pathway modu- lates ethanol teratogenesis. We have used CRISPR/Cas-9 At the most severe end of the FASD spectrum is Foetal to generate mutants altering mTORC1 function and are Alcohol Syndrome (FAS), where diagnosis is reliant on the currently assaying the ethanol sensitivity of these mutants. identifcation of a complex set of neurocognitive defcits and Disruption of the essential mTORC1 complex member rap- identifable facial features. To attain diagnosis, individuals tor sensitizes embryos to ethanol-induced defects, whereas would typically have been brought to the attention of care elevating mTORC1 function by disrupting tsc1a restores professionals or by guardians or parents at early stages of facial development in ethanol-treated pdgfra mutants and education when behavioural and neurodevelopmental def- heterozygotes. Te mTORC1 pathway is also associated with ciencies become apparent. Individuals with FAS facial criteria social behavior, and ethanol alters social behavior in zebrafsh. make up only a small proportion of those prenatally exposed Consistent with our fndings in the face, reduced tsc1a gene to alcohol, and cases are often missed. At the neonatal stage, dosage appears to protect against ethanol-induced social cognitive assesment is not possible, and an individual without behavioral reductions. Collectively, our fndings implicate the the characteristic facial features will not receive diagnosis mTOR pathway in multiple aspects of FASD. Tis work is untill later on in childhood. Previous studies in adolescent supported by NIH/NIDCR R01DE020884, NIH/NIAAA populations have utilised 3D imaging, accurately identifying and NIH/NIAAA U01AA021651 as a component of the FAS individuals, and objectively recognising subtle facial Collaborative Initiative on Fetal Alcohol Spectrum Disorders dysmorphism across the FASD spectrum. For this study, 3D (CIFASD) to J.K.E. images of infants taken at one month and one year were col- lected from a South African population, and neonatal images Ethanol and cannabinoids interact to induce were obtained from a Caucasian population from a clinic in birth defects through a mechanistic pathway Brighton, UK. Using surface-based analysis of facial form, we converging on primary cilia observe subtle changes in dysmorphism that occur at the two S.E. Parnell1, E.W. Fish1, K.E. Boschen1 (1 Chapel Hill, USA) time points. In addition, we identify subtle dysmorphology in those with a record of prenatal alcohol-exposure in the neo- Ethanol has long been known to be teratogenic, inducing natal population. At neonatal and infant stages, 3D imaging signifcant craniofacial and brain abnormalities during em- may accurately and objectively assist facial analysis in those bryogenesis. We have shown that exposure to both synthetic exposed prenatally to alcohol, particularly where features are cannabinoids and the natural cannabinoids found in canna- subtle and more challenging to identify. Early diagnosis is bis – THC and (CBD), during early gestation in paramount to providing early support and improving out- mice results in abnormalities similar to those caused by etha- comes. Supported by NIAAA Grant U01AA014809 as a nol, including midfacial hypoplasia, holoprosencephaly, and component of the Collaborative Initiative on Fetal Alcohol cleft lips/palates. Tese efects are synergistically produced by Spectrum Disorders (CIFASD). the combination of small doses of ethanol and cannabinoids. Te teratogenic efects of these drugs are mediated through reductions in the sonic hedgehog (Shh) signaling pathway Gut microbiota and alcohol: present and future within the neural tube and can be ameliorated in two ways: amplifcation of Shh signaling; or inhibition of cannabinoid Jose Antonio López-Moreno (Madrid, Spain) signaling with cannabinoid receptor 1 (CB1) antagonists. Ethanol inhibits Shh signaling through disruptions of pri- Gut microbiome may predispose to alcohol use mary cilia function (Shh requires cilia for normal signaling). We also show that cannabinoids inhibit Shh signaling disorder in rats 1 2 2 through inhibition of smoothened (Smo), the main efector Kshitij S Jadhav , Veronica L Peterson , John F Cryan , Benjamin 1 1 2 molecule of the Shh pathway, and CB1 binds to Smo in the Boutrel ( Lausanne, Suisse, Cork, Ireland) primary cilium, acting as an endogenous regulator to fne- tune Shh signaling and downstream mTOR pathway activity. Alcohol use represents a significant public health cost, Together, these data demonstrate a mechanistic pathway accounting for 4.5% of global disease burden. With current through which ethanol and cannabinoids act synergistically pharmacotherapies largely unsatisfying, discovering novel to impair embryonic development. Supported by NIAAA alternatives to prevent alcohol use disorder becomes a priority. grants R01AA026068 and U01AA021651 as a component Hence, identifying biological markers predicting vulnerability of the Collaborative Initiative on Fetal Alcohol Spectrum to develop excessive alcohol consumption may lead to a real Disorders (CIFASD) (S.E.P.). improvement of clinical care. With converging evidence sug- gesting that gut microbiota is capable of infuencing brain Utilising 3D facial analysis for the early and behavior, we aimed at investigating the gut microbiome identification of fasd associated facial in rats exhibiting uncontrolled alcohol seeking behaviors dysmorphism at neonatal and infant stages defned as: 1) inability to abstain during a signaled period

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 253 Congress of reward unavailability, 2) increased motivation assessed in lation of intestinal bacteria analyzed by Next Generation a progressive efortful task and 3) persistent alcohol seeking Sequencing and the similarities / diferences found between despite aversive foot shocks. Based on addiction criteria humans and animals. Acknowledgements: this work was sup- scores, rats were assigned to either Vulnerable or Resilient ported by the Ministerio de Sanidad, Consumo y Bienestar group. Not only Vulnerable rats displayed increased impulsive Social (Plan Nacional sobre Drogas 2018/050 to J.A.L.M) and compulsive behaviors, but also displayed increased relapse and the Fondo de Investigación Sanitaria (Red de Trastornos after abstinence and increased sensitivity to baclofen treat- Adictivos, FEDER, RD16/0017/0008 to J.A.L.M). ments compared to resilient animals. Following a 2-month wash out period, rats were sacrifced; dorsal striatum was Gut microbiota in alcohol related disease collected to assess dopamine receptor mRNA expression, Ali Keshavarzian1, Christopher B Forsyth1, Robin M Voigt1, and 16S microbiome sequencing was performed on caecal Garth R Swanson1, Faraz Bishehsari1, Maliha Shaikh1, Stefan contents. Analyses revealed signifcant correlations between Green1 (1 Chicago, USA) gut microbiome and impulsivity measures, as well as augmen- tations in striatal Dopamine 1 receptor (D1R) and reductions in D2R as vulnerability to AUD increased. Terefore, using The gut microbiota is currently one of the most active a singular translational approach based on biobehavioral research areas in medicine, including alcohol-related diseases. dispositions to excessive alcohol seeking without heavy However, this has not always been the case. Our laboratory intoxication, our observations suggests an association between has been a leader in investigating the complex relationship gut microbiome composition and the vulnerability to lose between the gut microbiota and diseases related to alcohol control over alcohol seeking behaviors. Acknowledgements: consumption/abuse. In early studies from our lab we demons- this work was supported by APC Microbiome Ireland trated that increased intestinal permeability to gut microbial and Science Foundation Ireland (SFI) [Grant No. 12/ contents was associated with development of alcoholic liver RC/2273], the Swiss National Science Foundation (Grant disease (ALD) in human patients. We then went on to carry No. 310030_185192) and the Department of Psychiatry, out studies in rats to show that disruption of the normal Lausanne University Hospital. KSJ is recipient of a Swiss pattern of the gut microbiota (so called dysbiosis) by chronic Government Excellence Scholarship. alcohol feeding was associated with intestinal hyperpermea- bility and ALD in those models. In addition we went on the Translational studies in alcohol-induced changes in show that dietary microbiota-directed therapy with probio- gut microbiota tics and prebiotic fber could reverse the changes in the gut Jose Antonio López-Moreno1, I. Rincón-Pérez1, V. Echeverry- microbiota as well as prevent intestinal hyperpermeability Alzate1,2, K. Bühler1, J. Calleja-Conde1, L. Segovia1, E. Giné1, and ALD. We then published the frst comprehensive study F. Rodríguez de Fonseca1,2, J. Albert1, J.A. Hinojosa1 (1 Madrid, of the gut microbiota in alcoholics with and without liver Spain, 2 Málaga, Spain) disease and found an association of specifc microbiota dys- biosis with ALD in human subjects. Our lab was also the frst Te intestinal microbiota is an own ecosystem within our one to show that disruption of circadian rhythms resulted body that evolved with us and that changes with our diet in dysbiosis of the gut microbiota. Signifcantly, we went on and other environmental infuences. In a sample of 507 to show that disruption of circadian rhythms together with university students we have found that the consumption of chronic alcohol feeding exacerbated intestinal hyperpermea- alcohol during weekends is associated with a change in the bility to gut microbial contents and resulted in increased composition of feces and intestinal bacteria. Te most plau- liver infammation and steatosis. Using the APC mouse sible hypothesis would be to consider that alcohol changes model of colon cancer, we showed that circadian disruption the intestinal microbiota although a second hypothesis can- increased chronic alcohol feeding promotion of colon polyps not be ruled out: that the diferences in the composition of and invasive polyps in that model and that microbiota direc- our gut microbiota would explain, at least in part, individual ted intervention with prebiotic fber feeding reversed these diferences in the amounts of alcohol consumed. To verify efects. We then went on the show that human shift workers this second hypothesis, we conducted a series of experiments with disrupted circadian rhythms exhibit dysbiosis and are with animal models (Wistar rats). A group of rats were sub- more susceptible (than day workers) to alcohol-induced gut jected to alcohol intoxications and another group of rats were leakiness with only moderate consumption of alcohol. Finally treated only with water. Tese two groups of rats were donors in our most recent studies we utilized intestinal organoids of fecal microbiota transplantation. Other groups of rats from chronic alcohol fed mice to show that decreased colonic received the fecal microbiota transplantation. Animals that microbiota short chain fatty acids (dysbiosis) is associated received the fecal microbiota transplantation from animals with epigenetic changes in colonic stem cell fate and junc- treated with alcohol drank more alcohol than animals that tional proteins resulting in intestinal hyperpermeability and received the microbiota transplantation from animals treated liver infammation. Our current studies are focused on further with water or treated with vehicle. We also found that the use investigating these circadian and intestinal stem cell efects of a cocktail of antibiotics to sterilize the intestine produced of the microbiota and alcohol related diseases and on how a reduction in alcohol consumption. During the presentation microbiota directed therapies can be used to treat and prevent we will show the main changes that we found in the popu- alcohol-related disease.

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Intestinal microbiota in alcoholic patients age of frst use. Tese fndings suggest that distinct regional 1 1 Dragos Ciocan ( Clamart, France) brain volumes in areas involved in learning and memory (hippocampus) and adaptive decision-making (frontal lobe), Chronic harmful alcohol consumption can induce a large and functional diferences in hippocampal efciency and sen- spectrum of conditions including liver, pancreatic, neuro- sitivity across emotional contexts may be important predictors logic and psychiatric diseases. Te intestinal microbiota is for the timing of alcohol use initiation. Characterizing brain recognized as an important player in the development and structure and function may therefore be useful in the search the severity of diferent diseases related to alcohol. Alcohol for biomarkers of risk for hazardous adolescent behaviors, induces changes in the composition and functions of micro- such as alcohol consumption. Further, such neurobiological biota (called dysbiosis). Also, alcohol increases the intestinal patterns, established early in the second decade of life, may permeability which allows translocation of bacteria, bacterial then escalate into adulthood, conferring neurobiological components, and bacterial metabolites (such as short chain risk of developing an alcohol use disorder later in life. fatty acids, bile acids and tryptophan metabolites) into the portal and the systemic circulation. Tese elements can reach Evaluating combined treatment versus single- the liver or the brain, establishing a gut-liver or gut-liver- focused treatment for depression and heavy brain axis, and can trigger local and systemic infammation. episodic drinking in college students Targeting microbiota has been shown to improve liver injury Paola Pedrelli1 (1 Boston, USA) both in animal models of diseases. Diferent strategies were tested: live bacteria (probiotics), microbiota transplantation, Heavy Episodic Drinking (HED) and depressive symptoms or the consumption of dietary fbres, such as pectin. All often co-occur among college students and are associated these methods can alter the ratio of bacterial species and thus with signifcant personal and societal problems. However, their functions. But although the connections between the evidence-based treatments are not available for these comor- microbiota and the host are well established, the underlying bid conditions. Te current study compared the efectiveness mechanisms, including key components that might serve as of a treatment combining Cognitive-Behavioral-Therapy potential therapeutic targets, remain to be elucidated. From a for Depression and Brief Motivational Interviewing (CBT- clinical perspective, well-designed studies that target micro- D+BMI) and CBT-D alone among 94 college students with biota in order to modify the clinical course, reverse disease depressive symptoms and HED. Both treatment programs or prevent complications are needed in patients with harmful were associated with signifcant reduction in HED, alcohol- alcohol consumption. related problems (ARP), and depressive symptoms, at the end of treatment and at one-month follow-up, of similar magni- tude. Moderators of outcomes were also examined. Among From alcohol initiation to disorder: perspectives students with fewer depressive symptoms at baseline CBT-D and treatment across the lifespan was associated with greater preservation of gains relative to CBT-D+BMI long-term. Furthermore, among students Marisa M Silveri (Boston, USA) with higher baseline frequency of HED, those who received CBT-D+BMI had a higher ARP reduction between baseline and the end of treatment than their peers receiving CBT-D. Adolescent neurobiology and the impact of While the study did not include a no-treatment condition, alcohol use initiation the magnitude of improvement during treatment was higher 1 1 Marisa M Silveri ( Boston, USA) than usually observed with time, suggesting a beneft of psychosocial treatments for this population. Depending on Early initiation of alcohol use is considered an important risk the main focus of treatment, HED or depression, diferent factor for the later development of an alcohol use disorder. approaches may be implemented. While behavioral and neurobiological alterations have been Mindfulness-based relapse prevention for drinking observed in relation to early onset alcohol use, it is not been fully elucidated whether diferences refect antecedents to, or reduction among community-dwelling older adults consequences of, early alcohol initiation. In our longitudinal seeking treatment for alcohol use disorder study, alcohol- and drug-naïve adolescents (n=81, 13-14 years- Katie Witkiewitz1 (1 Albuquerque, USA) old) completed neuroimaging at baseline, magnetic resonance imaging (MRI) and functional MRI during spatial navigation Treatments for alcohol use disorder (AUD) have progressed and emotional response inhibition task performance. Based considerably over the past 30 years. Yet, relatively few studies on quarterly follow-up assessments of alcohol use, those have been conducted to evaluate treatments that target drin- who initiated use by age 16 exhibited smaller hippocampal king reduction for less severe drinkers who are interested in volumes and larger posterior cingulate and superior frontal reducing alcohol consumption and even fewer studies have volumes than non-initiators. During fMRI, hippocampal examined treatments for older adults interested in drinking re- activation during memory retrieval was negatively correlated duction. Mindfulness-based relapse prevention is a treatment with age of frst use, while hippocampal activation during ne- for addiction that holds considerable promise for targeting co- gative response inhibition trials was positively correlated with gnitive and afective processes, and has recently been found in

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 255 Congress two randomized clinical trials to be superior to gold standard Precision medicine in alcohol dependence: cognitive behavioral treatment in reducing heavy drinking and matching phenotypes to pharmacotherapy drug use days. Yet, prior work as not examined mindfulness- Henry Kranzler1 (1 Philadelphia, USA) based relapse prevention (MBRP) for heavy drinking reduc- tions as primary outpatient treatment and has not examined Early eforts to identify predictors of response to alcohol MBRP in older adults. In the current study, we examined use disorder (AUD) treatment focused largely on patient the efectiveness of rolling group MBRP among treatment characteristics that moderate the response to psychotherapy. seeking individuals (n=84) with alcohol use disorder who were Although some signifcant moderators were identifed in interested in reducing their heavy drinking. We also examined Project MATCH, a large, multi-center trial that examined a the efects of treatment among older adults (age 60+, n=25). series of features as predictors of response to three diferent Results indicated a signifcant efect of treatment on drinking psychotherapies for AUD, the fndings have not substantially reductions. Te magnitude of drinking reductions also cor- altered clinical care. More recent eforts have examined gene- related with the number of MBRP group sessions attended. tic predictors of medication response, with the most widely Results from the subgroup analyses indicated the efects were studied gene variant-medication pair being the Asn40Asp also signifcant in those over the age of 60. Taken together, single nucleotide polymorphism in the mu-opioid receptor these results suggest drinking and craving reductions are gene as a moderator of naltrexone treatment response. In a achievable among people with AUD, including older adults. recent meta-analysis of that literature, we found no efect of Te current fndings also suggest a dose response relationship the Asp40 variant on the response to naltrexone. Mann et between MBRP sessions attended and drinking reductions. al. recently reported that a subgroup of individuals from the PREDICT Study identifed as reward drinkers showed an Pharmacological treatments for alcohol use disorder: 83% reduction in the likelihood of any heavy drinking when riding the waves of medications development treated with naltrexone compared to placebo. In a secondary Lorenzo Leggio1 (1 Baltimore and Providence, USA) analysis of a randomized trial of naltrexone among problem drinkers, reward drinkers treated with naltrexone reported Harmful alcohol use is a risk factor in more than 60 diseases signifcantly less frequent and less heavy drinking, and desire and injuries resulting in approximately 2.5 million deaths to drink mediated the efect of naltrexone on daily drinking. per year worldwide, therefore efective treatments for alcohol If validated prospectively, the reward drinker phenotype could use disorder (AUD) are needed. Among them, medications serve as a clinically useful self-report measure that predicts development represents a high priority in the alcohol feld. the subgroup of AUD patients who beneft most from nalt- However, only a few medications (“frst wave”) are currently rexone treatment. approved by the appropriate regulatory bodies (e.g., FDA, Harm reduction: an alternative avenue for alcohol EMEA) for AUD. Research has been conducted with the use disorder pharmacotherapy goal of testing medications already approved for other cli- Karl Mann1, Katie Witkiewitz1, the ACTIVE group1 (1 Mannheim, nical indications as potential efective treatments for AUD Germany) (“second wave”). Among several medications that may be mentioned in this group, two examples include GABAB receptor agonism via baclofen, a medication already approved Only 10-20% of individuals with a diagnosis of alcohol for spasticity; and alpha-1-blockade via and doxazo- dependence subscribe to a specifc treatment. Empirical data sin, two medications already approved for hypertension and show that the most important reason for this treatment gap is benign prostatic hyperplasia. More recently, additional eforts the requirement to subscribe to total abstinence. Te scientifc have been made toward the identifcation of new neurobio- discussion of this issue gained signifcant momentum when logical pathways involved in the development and mainte- the European Medicines Agency accepted Reductions in nance of AUD and possibly useful as new therapeutic targets alcohol consumption as an interim harm reduction strategy (“third wave”). Among them, neuroendocrine pathways such in the treatment of alcohol use disorder. Earlier studies have as ghrelin, GLP-1 and oxytocin seem promising and are found that signifcant reductions in alcohol use are common currently under investigation. Te overall goal of the clinical among individuals with alcohol use disorder, however it re- and translational research in the feld of medications develop- mained unclear whether reductions in drinking risk levels are ment for AUD is to increase the armamentarium of efective associated with signifcant improvements in health, quality of pharmacological treatments that health care providers may life, and other consequences of alcohol use disorder, and also use to treat patients with AUD and provide evidence toward whether reductions in drinking are stable over time. Te goal patient-oriented precision medicine. of our approach was to examine the correspondence between levels of alcohol consumption and experiences of drinking- related consequences, mental health, blood pressure, and liver The changing face of clinical trials for alcohol function tests during treatment among individuals receiving use disorders treatment for alcohol dependence in the COMBINE study. Results indicated reductions in WHO drinking risk levels were associated with significantly fewer alcohol related Henri-Jean Aubin (Villejuif, France) consequences, greater mental health, and improvements in

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 256 Congress physical health functioning, including reduced blood pres- and experience of negative afect/stress are expected to pre- sure and better liver function. Importantly, reductions in risk dict increased alcohol craving, which in turn leads to alcohol levels were also stable over time. Te results provide evidence use. Although our data EMA data failed to support expected of reductions in WHO risk levels as a viable alternative to intervention-specifc efects, we found independent efects of abstinence as an endpoint for alcohol clinical trials associated self-reported stress (2 items: stressed and overwhelmed), re- with meaningful reductions in alcohol related consequences ports of experiencing specifc stressors (fnancial/job-related, and improvements in mental and physical health. relationship, health-related, daily hassles), and negative afect on craving. For example, we found that relationship-related Placebo response in clinical trials for alcohol use stressors and daily hassles predicted level of craving at the disorders: can we improve clinical trial designs? within-subject level, whereas fnancial/job-related stressors Henri-Jean Aubin1, Claire Farina1 (1 Villejuif, France) and health-related stressors predicted level of craving at the between-subject level. Tese kinds of nuanced associations Te placebo response in Alcohol Use Disorder (AUD) trials between stress-related factors and alcohol-related outcomes has been shown to be negatively correlated to the treatment have yet to be examined using ecologically valid data. Further, effect size. This systematic meta-analytic approach gives we found independent efects for four out of fve cue exposure insight on how the choice of various endpoints and time measures. Specifcally, at the within-subjects level, we found points afect the placebo response in AUD trials. Moderator that exposure to alcohol advertising, seeing alcohol, being and meta-regression analyses can provide valuable informa- proximal to a drinking establishment, and hearing conver- tion regarding the impact of variables such as patient cha- sations about alcohol were all uniquely predictive of alcohol racteristics and trial design options on the placebo response. craving (though being around other individuals with whom Depending on endpoints and time points, placebo response they have consumed alcohol was not). efect size (standardized mean diference) may vary from va- Alcohol use among college students: lessons lues as high as 1.5 to 2.2. Return to any drinking at 12 weeks learned from the Protective Strategies Study Team was observed in more than 70% individuals in trials selecting 1 1 abstinent subjects at baseline. Placebo response moderators Protective Strategies Study Team ( Albuquerque, USA) difered according to endpoints. Nevertheless, journal impact factor appeared to be consistently related to a lower placebo Protective behavioral strategies (PBS) are specifc behaviors response. Interestingly psychiatric comorbidity did not appear one can utilize to minimize the harmful consequences of to signifcantly afect the placebo response efect size. Also, alcohol consumption. Tere has been an increasing amount studies published in recent years showed a higher placebo of interest in use of PBS among college students, especially response efect size for alcohol reduction-related endpoints, as an intervention target. In 2013, Pearson reviewed all PBS whereas earlier studies showed a higher placebo response studies conducted at that time. Tis review demonstrated efect size for abstinent-related endpoints. Some of these that despite the fact that a large number of studies have fnding can be valuable for future AUD randomized clinical been conducted on PBS, there have been few replication trials protocols. attempts. To advance the PBS feld, the Protective Strategies Study Team (PSST) was formed to conduct large multisite studies investigating PBS. In our frst study, we collected Examining psychosocial predictors of alcohol over 7,307 across 10 diferent states in the United States. In outcomes: implications for etiology, prevention, this presentation, we highlight our major fndings to date. Chiefy, we replicated and extended past fndings by exploring and treatment a wide range of more distal antecedents (i.e., mental health symptoms, drinking motives, impulsivity-like traits) that may Matthew R Pearson (Albuquerque, USA) afect alcohol-related outcomes via PBS use (i.e., mediation) as well as a number of factors that interact with PBS use to An ecological test of stress- and cue-induced predict outcomes (i.e., moderators). We also compare and contract alcohol PBS fndings with cannabis PBS fndings. alcohol craving Finally, we discuss the implications of these fndings for 1 1 1 Matthew R Pearson , Katie Witkiewitz , Eric Claus prevention and intervention eforts. (1 Albuquerque, USA) Beyond one size fits all: deviance regulation A sample of 68 participants were recruited and randomized theory-based interventions targeting use of to a mindfulness-based intervention (n=24), attentional bias alcohol protective behavioral strategies modifcation (n=21), or a health education/attentional bias Robert D Dvorak1 (1 Orlando, USA) assessment control condition (n=23). Our design included the collection of drinking data for 2 weeks via ecological The prototypical college student alcohol intervention is momentary assessment (EMA) before, during, and after the norm-based. Most of these interventions use personalized intervention. Our hypotheses were built from a craving-based normative feedback to correct college students’ overestima- model of drinking such that both exposure to alcohol cues tions of their peers’ alcohol use. Unfortunately, these inter-

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 257 Congress ventions tend to have small, short-lived efects on alcohol Do you feel your body or your emotions? use. Protective behavioral strategies (PBS) are behaviors one Drinking alcohol in the context of body-mind can engage in before, during, or after drinking to help reduce communication alcohol use, intoxication, and/or alcohol-related harms. PBS have been shown to be directly related to lower alcohol Marcin Wojnar (Warsaw, ) consequences even when controlling for levels of use, making it a promising intervention target. Tere is reason to suspect that traditional normative feedback will not translate well to Drink until you cannot feel feelings: the role of targeting PBS use. In this presentation, we review a social physical pain in predicting relapse to alcohol use psychological theory, deviance regulation theory (DRT), that following treatment for alcohol use disorder posits individuals engage in behaviors to stand out in positive Katie Witkiewitz1 (1 Albuquerque, USA) ways or avoid standing out in negative ways. We review the fndings from 5 studies conducted to date evaluating DRT- Identifying factors that predict a return to heavy drin- based interventions targeting alcohol PBS. Taken together, king (i.e., relapse) following alcohol treatment is critical these interventions have been shown to increase alcohol PBS for the development of relapse prevention interventions. use, which in turn reduces alcohol use and/or alcohol-related Physical pain is common among individuals with alco- problems. We discuss the implications of matching inter- hol use disorders (AUDs), yet few studies have examined vention content (in our case, positively framed vs. negatively associations between pain and alcohol relapse. Data from framed messages) to individuals’ preexisting beliefs (in our the COMBINE study (n=1383) and the United Kingdom case, their normative perceptions) in the broader context of Alcohol Treatment Trial (n=743) were used to examine the alcohol interventions. associations between physical pain and alcohol relapse in I am what i am: defining the nomological network randomized clinical trials for alcohol use disorders. Results of alcohol identity using meta-analysis indicated a signifcant association between physical pain Kevin S Montes1, Matthew R Pearson2 (1 Carson , USA, and alcohol relapse, which was mediated by experiences of 2 Albuquerque, USA) negative afect. Targeting acceptance and management of physical pain in the treatment of alcohol use disorder may help reduce relapse risk. Alcohol identity refers to the centrality of alcohol use to an individual’s self-concept (e.g., the extent to which alcohol Can you feel your own body? The role of use is important to one’s self-image or self/personal identity). interoception in alcohol use disorder Teory (e.g., PRIME theory of motivation; social identity Andrzej Jakubczyk1 (1 Warsaw, Poland) theory, theory of planned behavior) suggests that alcohol identity may serve as an important source of motivation that It has been suggested that interoception (which reflects guides alcohol use. Empirical research has also been conduc- the way one perceives somatic stimuli from the body) may ted to examine the associations between alcohol identity and contribute to alcohol use disorder (AUD) as it relates to the alcohol-related outcomes (e.g., frequency, quantity, conse- body’s experience of substance use or withdrawal. Moreover, quences, and dependence). Traditionally, alcohol identity there is growing evidence that interoceptive responses are has been assessed using self-report measures although more associated with immediate, discrete emotions. However, only recent investigations have assessed alcohol identity using a few studies have directly investigated associations between implicit measures (e.g., implicit alcohol identity). Over 40 interoception, emotion regulation and alcohol use. Te study studies have been conducted to examine the associations conducted in a group of sober alcohol-dependent individuals between alcohol identity and alcohol-related outcomes along showed that when controlling for level of anxiety, sleep pro- with narrative reviews that summarized fndings from many blems, age, sex and education, individuals with AUD scored of these studies. Missing from this extant literature is a signifcantly higher on self-reported interoceptive sensibility systematic review that includes a meta-analytic component and lower on interoceptive accuracy in comparison to healthy that synthesizes efect size estimates across these studies in controls. Moreover, in the group of subjects with AUD mea- order to derive a single, weighted efect size estimate between sures of emotional utilization remained a signifcant correlate alcohol identity and each alcohol-related outcome. In the of interoceptive accuracy, whereas lack of own emotional current study, we meta-analytically examined the associations awareness, difculties controlling impulsive behaviors when between alcohol identity and alcohol-related outcomes. We experiencing negative emotions, and appraisal of emotions also examined the associations between alcohol identity and remained signifcantly associated with interoceptive sensibi- known correlates of alcohol use (e.g., motives, expectancies, lity. Tese results have to be treated as preliminary and need negative afect, norms, cravings). In addition, we summarize to be replicated; however, fndings indicate that interocep- measurement approaches used in the assessment of alcohol tion may present a novel therapeutic target for treatment of identity. Finding from this meta-analytic study will inform AUD. a discussion surrounding opportunities to improve the mea- Can you feel what I am feeling – childhood surement of alcohol identity as well as how best to target alcohol identity in future prevention/intervention eforts. adversity, alcohol use and mentalization

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Maciej Kopera1 (1 Warsaw, Poland) Social cognition in severe alcohol-use disorders: from emotional decoding to dehumanization Although theoretical link between childhood trauma and experience mentalization has been established empirical evidence for it is still limited. Current data shows that the direction of this Pierre Maurage (Louvain-la-Neuve, Belgium) relationship might be individually shaped in selected at-risk populations. Childhood trauma is highly prevalent in treat- ment-seeking subjects with AUD and may play a signifcant It’s complicated: different tasks lead to different role in the development and severity of AUD. Te frst goal conclusions as to the severity of the impact of of the presented studies was to see if the presence of risky severe alcohol-use disorder upon social cognition alcohol use during the developmental age would infuence the Sharon Cox1 (1 London, UK) relationship between childhood adversity and mental states recognition in early adulthood (Study 1). Additionally, we Severe alcohol-use disorder (SAUD) is associated with wanted to see if the transgression from risky alcohol use to defcits in social cognition, frequently evidenced by errors in AUD would infuence the trauma-mentalization relationship emotional facial recognition and a lesser ability to theorise in another treatment seeking AUD sample (Study 2). Our about another’s state of mind. To date, little evidence exists fndings highlight an important and lasting role for variations on the clinical impact of these defcits, including how they in early life stress on individual diferences in adult social infuence treatment outcomes and are perceived by others. cognitive functioning. Findings from two studies will be presented; the frst study (N=123) highlights the impact of poor social cognition on Heavy drinking, depression, and treatment treatment outcomes (completion versus drop out), and how seeking for mental health concerns in physicians diferent tasks (binary versus non-binary responses) generate Kirk J Brower1 (1 Ann Arbor, USA) varying conclusions as to the extent of the defcits caused by SAUD. Results suggest that non-binary tasks which are Introduction: the relationship between heavy drinking and non-time defned allow adults with SAUD scope to explore their answers and self-correct, leading to lower error rates. In depression in physicians is worthy of further study. A large the second study (N-89), how apparent these problems are survey of U.S. physicians found that 15.3% screened positive to professionals who support these individuals is explored for an alcohol use disorder using the AUDIT-C, which through the correlation of estimates of defcits to experi- correlated signifcantly with depression. Te objective of mental data. Professionals’ ability to recognise these defcits this study was to look at the infuences of heavy drinking is contained to those who clients presenting with SAUD (HD) and depression on treatment seeking for mental and other clinical symptoms (e.g., anxiety, psychosis). Taken health concerns in physicians. It was hypothesized that each together, results suggest that the nature of the experimental would have independent efects and reduce the likelihood of tasks afects the degree to which we can estimate the severity treatment seeking. Methods: a faculty and physician health of the impact of SAUD upon social cognition. survey was sent by an email link to 3657 faculty members at a medical school. Anonymity was assured by using an external Self-evaluative emotions in severe alcohol use survey center, so that only de-identifed data were analyzed. disorders A total of 1710 (46.8%) of surveys were returned, including Delphine Grynberg1 (1 Lille, France) 1089 physicians who constituted the study group. Te sample consisted of 46% females with a modal age group between 36 Guilt and shame have been very little explored in severe to 45 (34.5%). Te NIAAA single question screen and crite- alcohol-use disorder (SAUD). Although previous fndings ria for HD, and the PHQ9 for depression were used. Results: suggest that the experience of shame may play a major role rates of any HD and for at least monthly HD during the past in the maintenance of SAUD, they do not allow to determine year were 23.1% and 7.3%, respectively. Moderate-to-severe (1) whether proneness to experience guilt and shame is asso- depression and any suicidal ideas during the past 2 weeks ciated with the experience of the emotions in the context of were endorsed by 13.6% and 3.7%, respectively. Physicians their consumption, (2) whether SAUD and healthy controls with at least monthly HD had signifcantly increased severity (HC) difer in terms of guilt and shame concerning their of depression. Frequency of suicidal thoughts were signif- alcohol consumption, and (3) whether these later are asso- cantly correlated with both any HD and at least monthly ciated with alcohol use severity. Te present study examined HD. Tose who drank heavily at least monthly were less these hypotheses in 40 patients diagnosed with SAUD willing to seek treatment for a mental health concern (60.8% according to DSM-V criteria and 54 HC (AUDIT scores<7). vs. 67.2%, p=0.02). Conclusion: frequent heavy-drinking Participants were instructed to complete the Test of Self- physicians had greater depression severity, but were less Conscious Afect-3, the Personal Feelings Questionnaire-2 willing to seek treatment for a mental health concern than (PFQ-2), the Hospital Anxiety and Depression Scale, the other physicians. Possible reasons for these fndings will be Alcohol Use Disorders Identifcation Test (AUDIT) and a discussed. questionnaire that has been developed for the present study,

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 259 Congress the “Substance Shame and Guilt” (SSG). Main results show Our model proposed that dehumanizing experiences would (1) PFQ-2-shame and PFQ-2-guilt are moderately asso- be associated with fundamental needs thwarting. Tese needs ciated with SSG-guilt and SSG-shame, (2) SAUD report are the psychological equivalent of thirst or hunger: shared by higher SSG-guilt and SSG-shame than HC and (3) SSG- all and causing important negative health consequences when guilt is positively associated with the AUDIT in SAUD only. thwarted. Additionally, it was hypothesized that dehumani- Tese results suggest that patients with SAUD experience zing experiences and fundamental needs thwarting would greater shame and guilt associated with their consumption together be linked to patients’ emotions, cognitions, and and that the consumption severity is associated with greater behaviors. Results supported that dehumanizing experiences guilt in SAUD. Tis study thus emphasized the importance were associated with increased fundamental needs thwarting. to consider shame and guilt in the maintenance of SAUD. Dehumanizing experiences were also associated with negative emotions, negative self-esteem, and dysfunctional coping Clinical impact of social cognition in treatment strategies, such as alcohol use. Our results suggest that dehu- seeking patients with severe alcohol-use disorders manizing experiences might play a crucial role in the vicious 1 1 Claudia Rupp ( Innsbruck, Austria) circle leading to SAUD.

Past years research witnessed growing evidence that cognitive defcits in patients with severe alcohol-use disorder (SAUD) Novel concepts in alcoholic liver disease include social cognition, most prominently defcits in (facial) emotion recognition. Until now, less is known about the cli- Sebastian Mueller (Heidelberg, Germany) nical relevance of these impairments in SAUD. In our pros- pective research studies, we were interested whether defcits Systemic inactivation of hypoxia-inducible factor in social cognition (1) contribute to less successful treatment prolyl 4-hydroxylase 2 in mice protects from outcome (relapse/dropout), and (2) recover “naturally” (with alcohol-induced fatty liver disease abstinence) during (about 8-week) SAUD treatment, or are 1 1 1 persistent cognitive problems. Main results of our studies are Anna Laitakari , Teemu Ollonen , Thomas Kietzmann , Gail 2 1 1 that (1) patients with SAUD presenting poorer (facial) emo- Walkinshaw , Daniela Mennerich , Valerio Izzi , Kirsi-Maria 1 1 1 tion recognition showed less successful treatment outcome Haapasaari , Johanna Myllyharju , Raisa Serpi , Elitsa Y with respect to relapse and/or early dropout. In addition, (2) Dimova1, Peppi Koivunen1 (1 Oulu, Finland, 2 San Francisco, USA) patients with SAUD showed no recovery with controlled abs- tinence during treatment in social cognition defcits, inclu- Alcoholic fatty liver disease (AFLD) is a growing health ding impaired (facial) emotion recognition ability, compared problem for which no targeted therapy is available. We with healthy controls. Our fndings evidence the clinical set out to study whether systemic inactivation of the main impact of social cognition defcits, particularly emotion reco- hypoxia-inducible factor prolyl 4-hydroxylase, HIF-P4H-2 gnition defcits on treatment success in SAUD. Moreover, (PHD2/EglN1), whose inactivation has been associated with our fndings indicate that clinically relevant social cognitive protection against metabolic dysfunction, could ameliorate it. defcits are rather persistent cognitive problems in SAUD HIF-P4H-2-defcient and wild-type (WT) mice or HIF- that would need special (e.g., neurocognitive rehabilitation) P4H inhibitor-treated WT mice were subjected to an ethanol treatments. New research focusing on the improvement of diet for 3-4 weeks and their metabolic health, liver and white these defcits in SAUD seems warranted. adipose tissue (WAT) were analyzed. Primary hepatocytes from the mice were used to study cellular ethanol metabo- Dehumanizing experiences of patients with severe lism. Te HIF-P4H-2-defcient mice retained a healthier alcohol-use: links with fundamental needs and metabolic profle, including less adiposity, better lipoprotein important clinical outcomes profle and restored insulin sensitivity, while on the ethanol Sullivan Fontesse1 (1 Louvain-la-Neuve, Belgium) diet than the WT. Tey also demonstrated protection from alcohol-induced steatosis and liver damage and had less WAT Dehumanization, the denial of one’s humanness, has infammation. In liver and WAT the expression of the key important negative consequences for social interactions. lipogenic and adipocytokine mRNAs, such as Fas and Ccl2, Dehumanization from the dehumanizer’s perspective has were downregulated, respectively. Te upregulation of meta- been widely studied in social psychology. However, victims’ bolic and antioxidant hypoxia-inducible factor (HIF) target perspective has been neglected. Moreover, despite dehuma- genes, such as Slcs 16a1 and 16a3 and Gclc, respectively, and nization being described as endemic in medicine, no study a higher catalytic activity of ALDH2 in the HIF-P4H-2- has investigated dehumanizing experiences (i.e the feeling defcient hepatocytes improved handling of the toxic ethanol of being dehumanized by others) in psychiatric populations. metabolites and oxidative stress. Pharmacological HIF-P4H To address these gaps, dehumanizing experiences of 120 inhibition in the WT mice phenocopied the protection patients with severe alcohol-use disorder (SAUD) were against AFLD. Our data show that global genetic inactiva- investigated. We argue that because patients with SAUD tion of HIF-P4H-2 and pharmacological HIF-P4H inhibi- are rejected and stigmatized against, two known antecedents tion can protect mice from alcohol-induced steatosis and liver of dehumanization, they might feel dehumanized by others. injury, suggesting that HIF-P4H inhibitors, now in clinical

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 260 Congress trials for renal anemia, could also be studied in randomized years; cirrhosis, 19%; HCC, 12%) patients were evaluated. 2) clinical trials for treatment of AFLD. Te survival and recurrence rates of HCC were examined in the patients of multi-center in Japan (532 ALD-HCC and Modeling of fibrosis pattern formation: from 209 NAFLD-HCC). Results: 1) Comparing with NAFLD, mouse models to human patients of chronic liver ALD were predominantly male and lower body mass index diseases and the complication of lifestyle-related diseases. As the 1,2 3,4 3 Seddik Hammad , Jieling Zhao , Jan G Hengstler , Sebastian genetic background, the ADH1B genotype GG and ALDH2 5 3,4 1 1 Müller , Dirk Drasdo , Steven Dooley ( Mannheim, Germany, genotype GG were observed more frequently and the MTP 2 3 4 5 Qena, Egypt, Dortmund, Germany, Paris, France, Heidelberg, genotype GG was decreased in ALD (ALD vs. NAFLD, Germany) ADH1B, 16% vs. 4%; ALDH2 84% vs. 44%; MTP 62% vs. 72%, respectively; all p<0.01). Comparing with and without Fibrosis is a consequence of repetitive liver injuries, e.g. HCC, the KCNJ15 genotype GG were identifed as the risk alcohol consumption. Liver fbrosis develops in diferent factors of ALD-HCC. 2) Te patients showed 5-year survival patterns according to the etiological factor i.e. pericellular to rates of ALD-HCC 43.7% vs. NAFLD-HCC 49.1%; 5-year septal pattern in alcoholic hepatitis. Te mechanism behind recurrence rates of 65.4% vs. 69.6 %, respectively. Conclusion: the generation of the diferent “scar” patterns is still elusive. the SNPs for the enzymes of alcohol metabolism were asso- Furthermore, this scar pattern is considered an early stage of ciated with ALD and the risk of diabetes were co-related to cirrhotic nodules in the advanced disease stage. We aim to ALD-HCC. Te survival and recurrence rates of HCC were defne (a) molecular driver(s) of fbrosis pattern formation. equally shown in ALD and NAFLD. We exposed mice to acute or chronically repeated doses (twice/week for 6 weeks) of CCl4, which like alcohol is The role of intestinal microbiota in alcoholic liver metabolized by CYP2E1, but presents with more severe disease liver damage in a shorter time frame. Spatial distribution of Ryuta Kitagawa1, Kazuyoshi Kon1, Maiko Suzuki1, Kenichi fbrosis formation and metabolic enzyme expression, namely Ikejima1 (1 Juntendo, Japan) CYP2E1, were analyzed in liver tissues. We found that reco- very of CYP2E1+ hepatocytes after acute CCl4 form exactly the same septal pattern of collagen scar walls that form upon Emerging attention has been paid for gut microbiota in the chronic injury, suggesting that CYP2E1+ hepatocytes are the human health and disease. Indeed, gut microbiota is dyna- trigger of the fbrotic pattern. To study this hypothesis, we mically altered by dietary factors, lifestyle, and alcohol intake. assumed a dynamic activator/inhibitor CYP2E1 mathema- Gut microbiota–dependent activation of hepatic innate tical model. Te current model already partially captures the immunity is important in the pathogenesis of steatohepatitis aforementioned CYP2E1/ECM pattern. We are currently caused by both alcohol and metabolic syndrome. Chronic challenging the model by mechanistic studies. Tese include alcohol exposure, as well as dietary overload, compromises gut i) crosstalk between activated/reverted HSCs and LSECs dif- barrier function causing increases in intestinal permeability, ferentiation; ii) communication between ECs lining the CV thereby aggravating translocation of bacterial products into and metabolically zonated hepatocytes; iii) testing a potential the portal blood. Pathogen-associated molecular patterns difusible inhibitor in the portal compartment, i.e. the bile (PAMPs) derived from gut microbiota elicit production and acid concentration. We are targeting the WNT/-Catenin release of infammatory cytokines through multiple innate pathway (CYP2E1 regulator) by monoclonal antibodies immune signaling pathways, resulting in the exacerbation against R-spondin1/2/3 in the fbrosed liver, and iv) test the of steatohepatitis. Te comorbidity of alcoholic liver disease mechanical role of ECM deposited by HSCs. Clinical appli- and metabolic syndrome has become an emerging clinical cation of the suggested model will be iteratively investigated problem worldwide. We have recently applied the mouse model of chronic-binge EtOH liver injury (NIAAA model) in F1, F2, F3 and F4 cohorts of patients with alcoholic liver y disease. for obese KK-A mice, mimicking alcoholic liver injury comorbid metabolic syndrome. For therapeutic approach, The characteristics of alcoholic liver disease in we investigated the efect of rifaximin (RFX), an oral non- Japan absorbed antibiotic, in this model. EtOH-feeding/binge Tomomi Kogiso1 (1 Tokyo, Japan) caused more severe hepatic steatosis, oxidative stress, and induction of infammatory cytokines in KK-Ay mice as com- Aim: the clinical features of alcoholic liver disease (ALD) pared to Bl/6 controls, which were markedly prevented by including the genetic background were not fully identifed in RFX treatment. RFX dramatically modifed the small intes- Japan. Here, we investigated that the clinical characteristics, tinal microbiota following chronic EtOH feeding, decreasing hepatocellular carcinoma (HCC), and the single nucleotide the relative abundance of the order Erysipelotrichales and polymorphisms (SNPs) associated with alcohol, glucose, and increasing the order Bacteroidales, without afecting EtOH- fat metabolism in patients with ALD compared to non- induced increase of net amount of viable bacteria. It is pos- alcoholic fatty liver disease (NAFLD). Methods: 1) ALD tulated that the modulation of small intestinal microbiota is (n=118; male, 86%; median age, 62 years; liver cirrhosis, critical for the prevention of alcoholic liver injury comorbid 58%; HCC, 31%) and NAFLD (n=200; male, 55%; age, 61 metabolic syndrome.

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Natural history of alcoholic liver disease: risk Alcoholic liver disease: impact of the type of factors, early disease detection and prognostic alcoholic beverage 1 1 markers Ina Bergheim ( Vienna, Austria)

Maja Thiele (Odense, Denmark) Alcohol intake is still among the leading causes of chronic liver diseases world-wide. Despite marked eforts made in many countries around the world to increase the awareness Epidemiology and natural history of alcohol- in the general population regarding the negative effects related liver disease on health associated with chronic and especially elevated Gro Askgaard1 (1 Copenhagen, Denmark) chronic intake of alcohol, the global proportion of the alcohol consumers has not markedly dropped throughout End-stage alcohol-related liver disease is a rare disease in the the last decades. Indeed, alcohol consumption still accounts general population (0.06% per year or lower in men > 45 years) for nearly 10% of global deaths among populations aged and knowledge of risk factors is needed to target screening 15-49. Epidemiological studies also suggest that per capita for early liver disease in individuals of high risk of the disease. consumption of alcohol from various alcohol beverages e.g. In this session we will focus on risk factors for alcohol- the beer, wine, spirts and other alcohol containing beverages related liver disease. We will discuss the infuence of alcohol including palm wine, or fermented beverages made of banana, amount (recent and earlier in life), alcohol drinking patterns sorghum or maze difers markedly between diferent areas (binge drinking/daily drinking, wine/beer/liquor, meal-related of the world. Epidemiological but also clinical and animal alcohol consumption) and age, smoking, and obesity as risk studies further suggest that diferent alcoholic beverages may factors of alcohol-related disease. Will cutting down on alco- impact the development of alcoholic liver disease diferenti- hol amount decrease the risk of further progression in liver ally. Tese fndings along with possible molecular mechanism disease? Individuals at particular high risk for end-stage liver involved will be reviewed and discussed. disease may be found at specialized alcohol treatment centers Vinyl chloride-induced interaction of nonalcoholic or as hospital patients given an alcohol problem diagnosis. and toxicant-associated steatohepatitis: protection Tese populations have a risk for end-stage alcohol-related liver disease of 7 to 16% after about 10 years. Moreover, some by the ALDH2 activator Alda-1 1 1 studies suggest half of patients with end-stage alcohol-related Juliane I Beier ( Pittsburgh, USA) liver disease had healthcare contacts in general practice or at the hospital before their diagnosis with obvious alcohol Te abundant environmental toxicant vinyl chloride (VC) problems. Tis indicates that there may be opportunities to shares similar metabolic pathways in the liver to alcohol. reach about half of patients with end-stage alcohol-related Specifcally, VC is metabolized via CYP2E1 and aldehyde liver disease with preventive interventions before diagnosis. dehydrogenase dependent pathways to produce the corres- ponding alcohol (chloroethanol, CE) and aldehyde (chloro- Current state-of-the-art prognostic and diagnostic acetaldehyde, CAA). VC causes steatohepatitis at high markers including genetic traits levels, but is considered safe at lower (i.e., sub-OSHA) 1 1 Maja Thiele ( Odense, Denmark) levels. However, we have previously shown that even lower VC levels exacerbate experimental fatty liver disease caused Tis talk will include an overview of the current best-in-class by high-fat diet (HFD). Mitochondrial oxidative injury non-invasive markers for diagnosis of fbrosis and prognos- and subsequent metabolic dysfunction appeared to play key tication in alcohol-related liver disease patients. Tese mar- roles in mediating this interaction. Mitochondrial aldehyde kers include ultrasound elastography techniques and serum dehydrogenase 2 (ALDH2) serves as a key line of defense markers that refect extracellular matrix formation, so called against endogenous and exogenous reactive aldehydes. Te direct fbrosis markers. Beyond the marker’s diagnostic role in current study therefore tests the hypothesis that allosteric an outpatient hospital setting, they may be used for case-fn- activation of ALDH2 with Alda-1 will protect against VC- ding or screening at a population level. However, due to the enhanced fatty liver disease. Mice were exposed to low VC lower prevalence of advanced fbrosis in at-risk populations, concentrations (<1 ppm), or room air for 6 hours/day, 5 days/ compared to patients referred to secondary healthcare, prese- week for 12 weeks, while on HFD or low-fat control diet lection of patients using risk scores may be used to increase (LFD). Some mice received Alda-1 (20 mg/kg i.p., 3×/week) true-positive rate and decrease false-positives, thereby mini- for the last 3 weeks of diet/VC exposure. Indices of liver mizing risk of overdiagnosis. Such risk scores are likely to injury, oxidative stress, metabolic and mitochondrial (dys) include genetic traits, co-occurance of metabolic risk factors, function were measured. As observed previously, low-dose alcohol drinking history and drinking pattern. VC did not cause liver injury in control mice; while liver injury caused by HFD was enhanced by VC. VC decreased hepatic ALDH2 activity of mice fed HFD. Alda-1 attenua- Pathophysiology of alcoholic liver disease ted oxidative stress, liver injury, and dysmetabolism in mice exposed to HFD+VC under these conditions. Importantly, Kenichi Ikejima (Chiba, Japan) alterations in mitochondrial function caused by VC and

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HFD were diminished by Alda-1. Previous studies have in- chronic injury, these changes likely contribute to activation dicated that liver injury caused by HFD is mediated, at least of a signifcant remodeling response that leads to scar for- in part, by enhanced mitochondrial autophagy (mitophagy). mation (i.e., fbrosis). Tis presentation will discuss some of Here, Alda-1 suppressed PINK1/PARKIN-mediated mito- the salient processes and players involved in the acute phase phagy. Taken together, these results support the hypothesis response of the ECM to liver injury after alcohol and other that ALDH2 is a critical defense against mitochondrial insults. injury caused by VC in experimental fatty liver disease. Te References ALDH2 activator Alda-1 conferred protection against liver 1 - Naba A, Clauser KR, Hoersch S, Liu H, Carr SA, Hynes RO. The matri- damage under these conditions, most likely via increasing some: in silico definition and in vivo characterization by proteomics of nor- clearance of aldehydes and preserving mitochondrial respi- mal and tumor extracellular matrices. Mol Cell Proteomics 2012;11:M111 014647. ratory function. 2 - Hynes RO. The extracellular matrix: not just pretty fibrils. Science The liver matrisome and inflammatory liver injury 2009;326:1216-1219. 3 - de Castro Bras LE, Ramirez TA, DeLeon-Pennell KY, Chiao YA, Ma Y, Dai in ALD Q, Halade GV, et al. Texas 3-step decellularization protocol: looking at the Gavin E Arteel1 (1 Pittsburgh, USA) cardiac extracellular matrix. J Proteomics 2013;86:43-52. 4 - Didangelos A, Yin X, Mandal K, Saje A, Smith A, Xu Q, Jahangiri M, et al. Extracellular matrix composition and remodeling in human abdo- Te strategic location of the liver between the intestinal minal aortic aneurysms: a proteomics approach. Mol Cell Proteomics tract and the rest of the body makes it a critical organ for 2011;10:M111 008128. clearance of xenobiotics and toxins that enter the portal 5 - Poole LG, Arteel GE. Transitional remodeling of the hepatic extracellular matrix in alcohol-induced liver injury. Biomed Res Int 2016;2016:3162670. blood. As the main detoxifying organ in the body, the liver has a high likelihood of toxic injury. It is therefore not Endoplasmic reticulum stress and oxidative stress surprising that the liver has tremendous ability to heal and regenerate from injury. Te complex and synchronized rege- in alcoholic liver injury comorbid metabolic nerative response in the liver can be perturbed and thereby syndrome can impact normal tissue recovery from injury or damage, Kazuyoshi Kon1, Maiko Suzuki1, Kenichi Ikejima1 (1 Tokyo, Japan) leading to progressive injury, and potentially liver failure. Te extracellular matrix (ECM) consists of a diverse range Te endoplasmic reticulum (ER) is a multifunctional orga- of components that work bi-directionally with surrounding nelle required for the regulation of calcium homeostasis, lipid cells to create a dynamic and responsive microenvironment metabolism, and protein synthesis. A number of cellular stress that regulates cell signaling, recruitment, and tissue function. conditions lead to the accumulation of unfolded or misfolded Te basic defnition of the ECM comprises fbrillar proteins proteins in the ER and disruption of the ER homeostasis, (e.g., , glycoproteins and proteoglycans). More re- which can trigger ER stress. ER stress activates the unfolded cently, groups have extended the defnition to include ECM protein response (UPR). Te UPR pathway includes induc- afliated proteins (e.g., collagen-related proteins), ECM re- tion of several molecular chaperones that restore cellular gulator/modifer proteins (e.g., lysyl oxidases and proteases) homeostasis by promoting the folding or degradation of and secreted factors that bind to the ECM (e.g., TGF and unfolded proteins; however, if ER stress is prolonged or too other cytokines); this broader defnition has been coined the severe, the signaling switches from pro-survival to pro-death, “matrisome” (1). Te ECM not only provides structure and leading to ER stress-induced apoptosis. Several studies have support for the cells in a tissue, but also acts as a reservoir for shown that ER stress contributes to the development of growth factors and cytokines and as a signaling mechanism alcoholic liver disease. Here we investigated the role of ER by which cells can communicate with their environment stress in chronic-binge ethanol (EtOH) model using obese and vise-versa (2). Quantitative and qualitative changes to KK-Ay mice. Chronic-plus-binge EtOH intake induced the ECM structure and superstructure can impact overall massive hepatic steatosis along with hepatocyte apoptosis health of the organ and organism. Remodeling of the hepatic and infammation, and increased ER stress markers including ECM/matrisome in response to injury is well understood binding immunoglobulin protein (Bip), unspliced and spliced in some contexts. For example, changes to the extracellular forms of X-box-binding protein-1 (uXBP1 and sXBP1, matrix associated with fbroproliferative diseases (i.e., fbro- respectively), inositol trisphosphate receptor (IP3R), and sis/cirrhosis) are considered almost synonymous with hepatic C/EBP homologous protein (CHOP), and also enhanced ECM changes. Proteomic-based studies in other organs oxidative stress markers heme oxygenase-1 and 4-hydroxyno- haves demonstrated that the matrisome responses dynami- nenal. Administration of 4-phenylbutyric acid, the chemical cally in composition after insult well before fbrotic changes chaperone, during chronic EtOH exposure ameliorated stea- to the organ (3, 4). Tese changes to the ECM may not alter tohepatitis after chronic-binge EtOH, and completely inhi- overall ECM architecture and are therefore histologically bited both ER and oxidative stress markers. Tese fndings undetectable. Nevertheless, these changes have potential indicated that binge EtOH intake after chronic consumption to alter hepatic phenotype and function (5). Tese acute induces massive ER stress-related oxidative stress followed by responses can be viewed as an arm of the wound healing liver injury, and inhibition of ER stress by chemical chape- response and facilitate recovery from damage, which resolves rone is a potential preventive therapy for alcoholic liver injury once the damage is repaired. However, under conditions of especially in obese subjects.

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Genetics of alcohol dependence hazards models determined whether the PRS increased risk of onset of hazardous drinking and dependence. In COGA, Joel Gelernter (West Haven, USA) the AUDIT-P PRS was strongly associated with alcohol dependence, symptom count, and maximum drinks, while The Genetics of antisocial personality disorder in AUDIT-C PRS was not an independent predictor of any phenotype. In ALSPAC, both PRS were strongly associated the context of alcohol dependence 1 1 with alcohol dependence, whereas AUDIT-P PRS was a Andrew McQuillin ( London, UK) superior predictor of maximum drinks. In GS, AUDIT-C PRS was a better predictor of weekly alcohol use, where- Antisocial personality disorder (ASPD) is characterised by as AUDIT-P PRS was strongly associated with CAGE impulsive, irresponsible and criminal behaviour. Tese per- scores. AUDIT-P PRS was also associated with ICD-based sonality traits begin in childhood or early adolescence and alcohol-related disorders in the UK Biobank sample. Lastly, continue into adulthood. Te prevalence of ASPD in the AUDIT-P PRS was associated with increased risk of onset general population is 2-3%, with estimates of 3% in men and of alcohol dependence in COGA, whereas AUDIT-C PRS 1% in women. Te rates are higher in certain populations was associated with increased risk of onset of hazardous with ASPD rates of 47% amongst male prisoners. ASPD drinking in ALSPAC. Our findings demonstrate that is highly comorbid with substance use disorders (SUD) and AUDIT PRS could dissect genetic infuences across alcohol studies have reported that 80-85% of individuals with ASPD use to misuse in both population-based and ascertained also meet criteria for a substance use disorder. Alcohol use cohorts. disorder in particular is highly comorbid with ASPD and in one study 71% of ASPD patients abused alcohol. We have Genetics of alcohol dependence in a family sample conducted a GWAS of ASPD symptom scores in two alco- Howard J Edenberg1 (1 Indianapolis, USA) hol dependence cohorts from the UK and the US (n=3,223). Tis analysis produced a genome wide signifcant fnding Tere is clear evidence that genetic variants afect the risk for a SNP located close to the SLCO3A1 gene on chromo- for alcohol dependence (AD), but few specifc variants have -08 some 15 (p=3.77×10 ). SLCO3A1 is a member of a family been identified to date. The Collaborative Study on the of organic anion transporter genes. Previous studies have Genetics of Alcoholism (COGA) conducted genome-wide reported association of this gene with AUD comorbid with association studies (GWAS) on AD, with secondary analyses bipolar disorder, with smoking behaviour and with inattentive looking at DSM-IV criteria endorsed. Te primary analyses symptoms in ADHD. Polygenic risk score analyses provided were among European Americans (EA), followed by trans- evidence for shared risk of ASPD in AUD subject with ancestral meta-analysis with an African American (AA) smoking behaviour, educational attainment and reproductive sample. In the GWAS of the EA sample, the functional traits. We report the frst genome-wide signifcant fnding SNP in ADH1B, rs1229984, was genome wide signifcant for ASPD and evidence for shared genetics risk for ASPD (GWS) for DSM-IV criterion count and 2 of the 7 criteria. in AUD across a range of behavioural traits. Trans-ancestral analysis strengthened the signal for criterion Polygenic contributions to alcohol use and alcohol count (p=2.6e-13) and 2 criteria, and elevated the signal use disorders across population-based and for DSM-IV AD to GWS. Other GWA fndings from the trans-ancestral GWAS were rs61826952 on chromosome clinically ascertained samples 1for AD, and rs7595960 on chromosome 2 for “time spent Emma Johnson1, Sandra Sanchez-Roige2, Arpana Agrawal1, 3 4 drinking”. Adding in data from several independent GWAS Toni-Kim Clarke , Alexis C Edwards , The Collaborative Study supported most of these fndings. A polygenic risk score on the Genetics of Alcoholism, The Avon Longitudinal Study (PRS) derived from the EA discovery GWAS signifcantly 1 2 of Parents and Children ( Saint Louis, USA, La Jolla, USA, predicted a small amount of variance in other EA datasets 3 4 Edinburgh, UK, Richmond, USA) (SAGE-EA, OZALC-EA). A promising endophenotype related to the risk for alcoholism is the sensitivity to alcohol Recent studies suggest that alcohol consumption and alcohol (SRE). Individuals who need to consume more alcohol to feel use disorders (AUD) have distinct genetic architecture. We its efects are at higher risk for heavy drinking and problems. examined polygenic risk scores (PRS) from a genome-wide GWAS and meta-analysis of SRE showed some novel asso- association study of the consumption and problem subscales ciations on chromosomes 6, 11, and 13. A PRS derived from of the Alcohol Use Disorders Identifcation Test (AUDIT-C the EA SRE signifcantly predicted alcohol dependence and and AUDIT-P) in four independent samples: an ascer- criterion count in the independent SAGE-EA subset, and tained cohort, the Collaborative Study on the Genetics of one from the AA SRE did in the SAGE-AA subset. Further Alcoholism (COGA); and three population-based cohorts, progress will require substantially larger samples, including the Avon Longitudinal Study of Parents and Children non-European populations, and will beneft from detailed (ALSPAC), Generation Scotland (GS), and a subset of the characterization of AD and its symptomology. UK Biobank (UKB). Regression models examined the cor- Alcohol GWAS results in different populations – relation between the AUDIT-C and AUDIT-P PRS and a variety of alcohol-related phenotypes. Cox proportional AA, EA, Asian – and attendant implications

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1 1 1 2 Joel Gelernter , Hang Zhou , Daniel Levey , Henry Kranzler , glands or pituitary tissues were processed for determination 3 1 2 3 Murray Stein ( West Haven, USA, Philadelphia, USA, San of biochemical changes and histopathologies for tumor Diego, USA) characterization. In the case of mammary tumor develop- ment, overall tumor multiplicity was greater in the ofspring Alcohol-related phenotypes – alcohol use disorder, quanti- from the alcohol-fed group compared to the control groups, ty-frequency measures such as AUDIT-C, and maximum indicating a decrease in tumor latency. Alcohol-exposed ani- habitual alcohol use – are moderately heritable; as is the case mals developed more malignant tumors and more for other complex traits, the specifc risk variants and overall receptor--negative tumors relative to the control groups. In genetic architecture difer somewhat between populations. In the case of prostate tumorigenesis, prenatal alcohol-exposed some cases, as for the trait of “maximum habitual alcohol use,” rats showed histological evidence for high-grade prostatic the interpopulation diferences are very useful in a practical intraepithelial neoplasia (PIN) primarily in the ventral pros- sense for fne mapping of risk variants, because of diferences tate, whereas control animals showed only low-grade PIN. in linkage disequilibrium between populations. At least one Prenatally ethanol-exposed rats treated with carcinogen and high odds ratio protective variant is almost entirely specifc to testosterone also showed increased number of proliferative certain Asian populations (ALDH2 rs671). Notwithstanding cells and androgen receptor with concomitant decreased these diferences and the general recognition of the impor- levels of tumor suppressor proteins in the ventral prostate. tance of studying major world populations, the available cha- Our results also show that pituitaries of fetal alcohol-exposed racterized samples for European-ancestry subjects are much rats upon estrogen challenge developed -secreting larger than those for African- and Asian-ancestry. Te US tumors (prolactinomas) that were hemorrhagic and often Million Veteran Program (MVP) presently includes samples penetrated into the surrounding tissue. Pituitary tumors of of European-ancestry subjects in the hundreds of thousands, fetal alcohol-exposed rats produced higher levels of hemor- and African-ancestry, in the tens of thousands. Te largest rhage-associated genes and proteins and multipotency genes reported studies of Asian ancestry barely reach the thousands. and proteins. Cells of pituitary tumor of fetal alcohol exposed Tis presentation will review available results in these three rat grew into tumor spheres in ultra-low attachment plate, populations; emphasize knowledge that has been gained only expressed multipotency genes, formed an increased number of through the study of multiple populations; and prospects for colonies, showed enhanced cell migration, and induced solid the future. tumors following inoculation in immunodefcient mice. Tese data suggest that fetal alcohol exposure programs some of Epigenetic effects of alcohol exposure in brain the endocrine tissue to develop aggressive tumors. Although the exact mechanism for the tumor promotion efect of fetal and in blood: an implication of methylation alcohol is not clearly established, but our preliminary studies biomarker for alcohol use disorder suggest the possibility that fetal alcohol programs some of these endocrine cells acquire stemness that enhances neoplas- Ke Xu (New Haven, USA) tic properties for developing aggressive tumors. Genome-wide DNA methylation in PFC of AUD Fetal alcohol exposures promote the development subjects: insights on the epigenetic regulation of of aggressive tumors in the endocrine glands the glucocorticoid receptor Dipak K Sarkar1 (1 New Brunswick, USA) Eleonora Gatta1, Dennis R Grayson1, James Auta1, Vikram Saudagar1, Erbo Dong1, Ying Chen1, Harish R Krishnanv1, Tere have been several studies demonstrating that alcohol Jenny Drnevichv1, Subhash C Pandey1, Alessandro Guidotti1 abuse promotes development of aggressive tumors in breast, (1 Chicago, USA) prostate, pancreas, and colon tissues in human patients. Whether fetal alcohol exposures promote development of Individual vulnerability to develop psychiatric disorders aggressive tumors in the ofspring during adult period are depends on an intricate interplay between the genetic not well studied. Using rat animal model of fetal alcohol background and the environment. Environmental factors, exposure, we studied the susceptibility of the growth of including substance abuse and stress, cause long-lasting aggressive tumors in the mammary, prostate and the pitui- changes in the regulation of gene expression in the brain via tary glands during the adult period. Pregnant laboratory rats epigenetic mechanisms, such as DNA methylation. Similar were fed between gestational days 7 and 21 with a liquid to stress, alcohol stimulates glucocorticoids release that bind diet containing alcohol, pair-fed with isocaloric liquid diet, to specifc receptors, i.e., the glucocorticoid receptor (enco- or fed ad libitum with rat chow. Between 50 to 90 days of ded by NR3C1). Te human NR3C1 gene is comprised of age, fetal alcohol-exposed and control rats were given a dose nine untranslated alternative first exons (1A-J) and eight of N-Nitroso-N-methylurea (NMU) to induce mammary translated exons (2 to 9). Seven of the exons 1 variants are cancer growth in female ofspring, NMU and testosterone to embedded within a CpG island known to be susceptible to induce prostate tumor in male ofspring, or ovariectomized epigenetic regulation via DNA methylation. Tese epigene- and implanted with an estrogen capsule to induce pitui- tic changes have been associated with psychopathological tary tumors in female ofspring. Mammary glands, prostate conditions in adulthood. However, little is known on the

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1 1 1 role of DNA methylation mechanisms in the expression of Robert Philibert , Meeshanthini Dogan , Jeffrey Long , James 1 stress-responsive genes in the brain of alcohol use disorders Mills (Coralville, USA) (AUD) subjects. Using a genome-wide DNA methylation approach (Infnium® MethylationEPICBeadChip, Illumina) Background: alcoholism is the third largest preventable followed by the identifcation of 5-methylcytosine (5mC) and cause of morbidity and mortality. Uniquely, in the absence 5-hydroxymethylcytosine (5hmC) by specifc immunoassay, of acute intoxication, there is no readily employable test for we identifed a diferential pattern of DNA methylation in assessing unhealthy levels of alcohol consumption. In 2014, AUD. Post-mortem brain samples were obtained from 25 we published the frst genome wide study of heavy alcohol controls and 25 AUD subjects from the New South Wales consumption (HAC). In this presentation, we expand on Tissue Resource Centre (University of Sydney, Australia). those earlier fndings using data and biomaterials (both saliva Bioinformatic analyses of diferentially methylated genes and whole blood DNA) from 143 participants with current highlight biological processes containing genes related to HAC and 200 abstinent controls. Results: using DNA from stress adaptation, including NR3C1. We validated some of whole blood, we show that a set of four methylation sensitive these data and observed that chronic alcohol drinking results digital PCR assays have a Receiver Operating Characteristic in a signifcant increased methylation of the NR3C1 exon (ROC) area under the curve (AUC) of 0.96 for detecting variant 1H, with a particular increase in the levels of 5hmC those with HAC using DNA from whole blood with similar over 5mC. Tese changes in DNA methylation were associa- fndings being obtained. After a mean of 21 days of inpa- ted with reduced NR3C1 mRNA and protein expression in tient enforced abstinence, methylation status at one of these PFC as well as other cortico-limbic regions of AUD subjects markers, cg04987734, began to revert to baseline values. Re- when compared to controls. Furthermore, we show that examination of methylation data from our 2014 study with the expression of several stress-responsive genes (e.g., CRF, respect to this locus demonstrated a similarly signifcant re- POMC, FKBP5) is altered in the PFC and hippocampus of version pattern at cg04987734 in association with treatment AUD subjects. Tese data suggest that alcohol-dependent enforced abstinence. When the saliva DNA is used in place aberrant DNA methylation of NR3C1 and consequent of whole blood, similar fndings with respect to AUC and changes in other stress-related genes might be fundamental methylation reversion are observed. Conclusions: we conclude in the pathophysiology of AUD and lay the groundwork that clinically implementable dPCR tools using DNA from for treatments targeting the altered epigenetic regulation of blood or saliva can sensitively detect the presence of HAC NR3C1 in AUD. Supported by the P50AA022538 NIAAA- and monitor alcohol treatment results. Tese digital PCR NIH grant to SCP and AG. tools will be useful to clinicians and researchers in monitoring those enrolled in substance use disorder treatment, employee DNA-methylation abundantly associates with fetal wellness programs and insurance underwriting. alcohol spectrum disorder and its sub-phenotypes I.M. Krzyzewska1, J.M. Cobben1, A. Venema1, A.N. Mul1, A. Polstra1, A.V. Postma1, R. Smigiel2, K. Pesz2, J. Niklinski3, M.A. Novel alcohol-induced epigenetic signaling: Chomczyk3, P. Henneman1, M.M.A.M. Mannens1 (1 Amsterdam, neuroimmune genes and miRNAs The Netherlands, 2 Wroclaw, Poland, 3 Bialystok, Poland) Fulton T Crews (Chapel Hill, USA) Aim: Fetal Alcohol Spectrum Disorder (FASD) involves prenatal growth delay, impaired facial and central nervous system development and causes severe clinical, social-eco- Neuronal-glial signaling through miRNA let7 nomic burdens. Here we aim to detect DNA-methylation and Toll-like Receptor 7 induce interferons and aberrations associated with FASD and potential FASD negative affect diagnostic and prognostic biomarkers. Patients and methods: L.G. Coleman1, J. Zou1, L. Qin,1 S.S, Moy1, F.T. Crews1 (1 Chapel FASD diagnosis was established according to golden- Hill, USA) standard protocols in a discovery and independent replication cohort. Genome-wide diferential methylation association Neuroimmune activation is a prominent feature of alcoholic and replication analyses were performed. Results: we identi- neuropathology. Tis includes activation of neuroimmune fed several loci that were robustly associated with FASD or Toll-like Receptors (TLRs) with release of their endogenous one of its sub-phenotypes. Our fndings were evaluated using agonists. Neuronal-glial interactions likely mediate neuroim- previously reported genome-wide surveys. Conclusions: we mune responses to alcohol, however specifc intercellular have detected robust FASD associated DMPs and DMRs signaling mechanisms associated with alcoholic behavioral for FASD in general and for FASD sub-phenotypes, i.e. on pathology need to be identifed. We found a novel glial to growth delay, impaired facial, and CNS development. neuronal signaling pathway, involving TLR7 and its endo- genous agonist, miRNA let-7, which contributes to neuronal A rapid methylation sensitive digital PCR test cell death, interferon (IFN) induction, and negative afect. that can sensitively and specifically assess Ethanol causes the secretion of let-7 in media microvesicles heavy alcohol consumption and monitor alcohol from microglia and astrocytes, which can activation TLR7 treatment using DNA from blood or saliva in neurons. In postmortem human alcoholic cortex, TLR7

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 266 Congress and its downstream signaling components IRF7 and IFNs persistent AIE-induced neuropathology through epigenetic were induced. In mice, chronic binge ethanol induced let-7, mechanisms that can be reversed in adulthood. TLR7, and pIRF7, and sensitized to immune responses and Glia-to-neuron transfer of inflammatory degeneration due to the TLR7 agonist Imiquimod (IMQ). proteins and miRNAs via extracellular vesicles: In primary ex-vivo slice culture, ethanol caused secretion of a new mechanism underlying ethanol-induced miRNA let-7 in media microvesicles, and inhibition of TLR7 with siRNA or a novel small molecule antagonist prevented neuroinflammation 1 1 1 1 1 ethanol-induced cell death. Te TLR7 antagonist also bloc- F. Ibáñez , M. Pascual , J. Ureña , C. Guerri ( Valencia, Spain) ked neurodegeneration to chronic binge ethanol in mice. Using immortalized neuronal (SH-SY5Y), microglial (BV2), Extracellular vesicles (EVs) participate in intercellular signa- and astrocyte (U373MG) cell lines, and conditioned media ling, and in the regulation and amplifcation of neuroin- transfers, we found ethanol induced glial secretion of miRNA fammation. We have shown that ethanol activates glial cells let-7 and IFN induction in neurons. Microglial depletion through Toll-like receptor 4 (TLR4), triggering neuroin- ex-vivo blunted ethanol or IMQ-TLR7 induction of TNF fammation. We evaluate if ethanol and the TLR4 response and IL-1, but not IFNs suggesting neuronal or astrocytic change the release and the infammatory content of astro- responses. In a 5-week binge model, ethanol caused persistent cytes-derived EVs, and whether these vesicles are internalized negative afect (anxiety-like behavior, and conditioned fear by neurons, spreading neuroinfammation. Cultures of corti- memory), with IFN expression correlating with persistent cal neurons and astrocytes were used. EVs were isolated from conditioned fear memory. the extracellular medium of WT and TLR4-KO astrocytes, treated or untreated with ethanol (40 mM) for 24 h. Te EVs Neuroimmune and epigenetic mechanisms regulate content in infammatory proteins, mRNA and miRNAs was adolescent binge ethanol-induced loss of basal analyzed by Western blot and RT-PCR in astrocyte-derived forebrain cholinergic neurons and hippocampal EVs and in neurons incubated or not with these EVs. A neurogenesis functional analysis of the miRNAs was also performed. We 1 1 Ryan Vetreno ( Chapel Hill, USA) show that ethanol increases the number of secreted nanove- sicles and alters their content by raising the levels of both Adolescence is a critical period of neurotransmitter system infammatory-related proteins (TLR4, p65, IL-1R, caspase-1, refnement and neuroplasticity that coincides with develop- NLRP3) and miRNA (mir-146a, mir-182 and mir-200b) in ment of adult cognition. Binge drinking and alcohol abuse the EVs from WT-astrocytes compared with those from the are common during adolescence causing lasting patho- untreated WT cells. Ethanol did not change the number and logy. We fnd adolescent intermittent ethanol (AIE), which the content of TLR4-KO astrocytes-EVs. We further showed models human binge drinking, persistently decreases basal that astrocytes’ EVs were internalized by naïve neurons, chan- forebrain cholinergic neuron (BFCN) populations and hip- ging their physiological functions and increasing infamma- pocampal neurogenesis. Reciprocal connections between the tory markers and miRNAs (e.g. mir-146a) levels, along with basal forebrain and hippocampus are critical for maintenance miRNAs-target genes (Traf6, Mapk14, Stat1 and Foxo3) in of BFCNs and neurogenesis. We linked proinfammatory si- neurons treated with ethanol-treated WT astrocyte-derived gnaling to loss of BFCNs and neurogenesis as treatment with EVs. Tese results suggest that astrocyte-derived EVs could lipopolysaccharide mimics and anti-infammatory treatments act as cellular transmitters of infammation signaling by prevent AIE pathology. An imbalance between neuroim- spreading and amplifying the neuroinfammatory response mune/neurotrophic signaling might contribute to persistent induced by ethanol through TLR4 activation. AIE-induced loss of BFCNs and neurogenesis through Role of microRNA 137 mediated changes in histone epigenetic mechanisms. In the basal forebrain, AIE increases histone 3 lysine 9 dimethylation (H3K9me2) occupancy at methylation in adolescent alcohol-induced anxiety Chat and Trka gene promoters, which is associated with and alcohol drinking behaviors in adulthood silencing of gene transcription. Exercise and acetylcholin- Subhash C Pandey1, Evan J Kyzar1, J. Peyton Bohnsack1, Huaibo esterase inhibitor (AChEI) treatment post-AIE reversed the Zhang1 (1 Chicago, USA) loss of BFCNs, and restored the neuroimmune/hippocampal NGF balance and increased histone methylation. Tese data Binge drinking during adolescence increases risk for psy- suggest AIE-induced loss of BFCNs might involve silencing chiatric disorders later in life. Epigenetic mechanisms such of cholinergic phenotype and not cell death. Similarly, we as microRNAs (miRNAs) may contribute to this increased fnd AIE increases H3K9me2 and reduces H3K9 acetylation risk via molecular changes in the amygdala. Here, we inves- in the adult hippocampus, the latter associated with dimi- tigated the role of miR-137 and its targeting of the epige- nished BDNF expression. Treatment with an AChEI and netic enzyme lysine-specifc demethylase 1 (LSD1) in the the histone deacetylase inhibitor TSA post-AIE reversed adult amygdala after adolescent intermittent ethanol (AIE) the AIE loss of hippocampal neurogenesis, and restored the exposure. Rats were exposed to 2g/kg ethanol (2 days on/of; neuroimmune/BDNF neurotrophic balance and epigenetic AIE) or intermittent n-saline (AIS) during postnatal days modifcations. Together, these data suggest that an imbalance (PND) 28-41 and allowed to grow to adulthood for analysis of neuroimmune/neurotrophic signaling might contribute to of behavior and biochemical measures. Some adult rats were

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 267 Congress cannulated in the central nucleus of amygdala (CeA) and comorbidities and evolution, circuit care, anatomic and func- infused with miR-137 antagomir with or without concur- tional brain imaging (IRM, 18-FDG-PET), nutritional sta- rent Lsd1 siRNA infusion prior to analysis. AIE increases tus, CSF biomarkers (tau, beta-amyloid peptids, neurogranin, miR-137, decreases Lsd1 expression, and decreases LSD1 neuroflament light chain). We will insist on the prevalence occupancy at the brain-derived neurotrophic factor exon of multifactorial origin of cognitive disease in this population. IV (Bdnf IV)promoter in adult amygdala. Infusion of miR- For the patients with strictly alcohol-related cognitive disor- 137 antagomir into the CeA rescues AIE-induced alcohol der, we will identify items correlated with a better prognosis drinking and anxiety-like behaviors. miR-137 antagomir of this disease. infusion in the CeA also normalizes the decreased Lsd1 Neuropsychological Impairments and brain expression, decreased LSD1 occupancy, and decreased Bdnf alterations in “uncomplicated” patients with IV expression due to increased H3K9me2 occupancy seen in Alcohol Use Disorder the amygdala of AIE adult rats. Finally, co-infusion of Lsd1 1 1 1 siRNA into CeA prevents the miR-137 antagomir-induced Anne-Lise Pitel , Alice Lanièpce , Nicolas Cabé , Laurent 1 1 1 1 rescue of molecular changes and anxiety-like behaviors. Tese Coulbault , Géraldine Rauchs , Shailendra Segobin ( Caen, results suggest that increased miR-137 in the CeA play an France) important role in chromatin remodeling and adult psycho- pathology caused by adolescent alcohol exposure. Supported Alcohol Use Disorder (AUD) is associated with altered brain by the NIAAA-NIH UO1AA-019971, U24AA-024605 & structure and function well before the development of neuro- P50AA022538 grants and senior VA career scientist award logical complications. Two brain circuits are mainly afected in to SCP and F30AA024948 to EJK. “uncomplicated” AUD patients: the frontocerebellar circuit in- volved in motor abilities as well as working memory and exe- cutive functions; and the Papez circuit implicated in episodic Alcohol-related cognitive impairment (ARCI): memory. However, the pathophysiology of the brain dysfunc- Korsakoff, Alzheimer, and friends tion observed in AUD remains unclear since the nature and extent of cerebral damage and cognitive defcits do not seem Florence Vorspan (Paris, France) to be directly related to the duration or severity of the alcohol history. Other factors including associated malnutrition po- tentially resulting in altered thiamin metabolism or defciency, Epidemiology: early-onset dementia is mostly liver disease, or sleep disturbances may favor the development related to alcohol use disorders or exacerbate alcohol-related neuropsychological defcits and Michaël Schwarzinger1 (1 Paris, France) brain abnormalities. Te role played by these factors must be further investigated since they are potential therapeutic tar- We conducted a nationwide retrospective cohort of all adult gets that could prevent or reduce brain dysfunction in AUD. (≥20 years) patients admitted to hospital in metropolitan Biomarkers in ARCI: what can we learn from the France between 2008 and 2013. Of all 81,958 cases of early- past 20 years of Alzheimer’s disease research and onset dementia (<65 years), 39,206 (47.8%) were related care? to alcohol use disorders. Early-onset dementia was more 1 1 frequently recorded in men (51,339 [62,6%]) than women Emmanuel Cognat ( Paris, France) (30,619 [37.4%]), with stronger association with alcohol use disorders (29,944 [58.3%] vs. 9,262 [30.2%]). Tis nationwide Cognitive neurodegenerative disorders are slowly progressive study suggests that the burden of early-onset dementia could brain diseases with overlapping clinical features and limited be substantially alleviated by reinforcing alcohol policies. access to brain pathology in living patients. Tus, important eforts have been made during the past decades to develop Clinical care: diagnosis and prognosis of Wernicke- diagnostic biomarkers that reflect pathological processes Korsakoff disease and severe alcohol-related and prognostic markers that could predict the course of cognitive impairment: Alcomemo cohort the disease. Research in this feld has been most active in Julien Azuar1, Frank Questel1, Virgile Clergue-Duval1, Thomas Alzheimer’s disease, the most frequent cause of cognitive dis- Barré1, Florence Vorspan1 (1 Paris, France) turbance in older patients but recent advances have been made in the past few years in other neurocognitive disorders such as Cognitive disorders are common in patients with alcohol- frontotemporal lobar degeneration. Alcohol-related cognitive related disorders (AUD). Tey must be screened and charac- impairment (ARCI) is a complex condition with frequent terized in order to obtain appropriate care. An observational multifactorial origin and difculties to predict prognosis. cohort of patients with AUD and severe cognitive impair- Co-existing neurodegenerative pathology does not seem rare. ment is being constructed from 2013 in our Addictology Tus there is a crucial need for both diagnostic and prognos- Department, with the help of a multicenter network called tic biomarkers useable in patients with ARCI. Development Resalcog. Tis network helps to keep a patient of alcohol of such biomarkers may take advantage of the lessons learnt for several months. We will describe the characteristics of from past and current research and use in clinical practice of this cohort of 124 patients, including clinical description of biomarkers in AD and other neurodegenerative disorders.

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 268 Congress

Alcohol use disorders in context of dual who drink alcohol develop AUD. Importantly, the mecha- diagnosis: did DSM make us lose the MATCH? nisms underlying loss of control over alcohol use and an individual’s risk to develop AUD remain incompletely under- Georges Brousse (Clermont-Ferrand, France) stood. Age of onset of alcohol use and certain personality traits are thought to play an important role in the risk for Dimensional perspectives for a pragmatic AUD. Neurobiologically, exaggerated involvement of the dor- therapeutic approach solateral striatum (DLS) has been proposed to contribute to Maurice Dematteis1 (1 Grenoble, France) habitual alcohol seeking and loss of control over alcohol use. We assessed the contribution of these factors to the propen- sity to lose control over alcohol seeking in rats. Our studies Dual disorders are common and polydrug use is the norm. showed that conditioned suppression of alcohol seeking, as a In patients combining both, the clinical presentation results measure for alcohol use in the face of adversity, was less pro- from a mixture that makes a categorical diagnosis as well as nounced in rats with adolescent-onset alcohol consumption, a specifc and stepped care difcult to establish. However compared to adult-onset animals. Further, rats that show complex the clinical picture, its deconstruction by elementary high levels of social play behaviour as juveniles consumed functional dimensions and endophenotypes (e.g. impulsivity) more alcohol but showed intact conditioned suppression allows for pragmatic, gradual and integrative holistic treat- of alcohol seeking, unlike rats that displayed low levels of ment suited to the patient’s needs, resources and ecology. social play, refecting a lack of control over alcohol seeking. Tere are currently no validated strategies in the literature. Furthermore, preliminary analysis of optically-induced exci- According to our experience and the Research Domain tatory neurotransmission in cortical-DLS projection neurons Criteria approach, we propose a framework that provides showed increased facilitation of paired-pulse responses in the functional understanding (drug’s functions in the psychic DLS in rats with a high alcohol drinking phenotype. Tese economy) and treatment of addictions which are considered data suggest a complex relationship between age of onset, to be dysfunctional adaptive strategies. Motivation- and edu- social development and DLS plasticity the development of cation-based treatment aims at restoring functional autonomy AUD-like behaviour in rats. and quality of life in accordance with the patient’s needs, and combines dimensional pharmaco-psychotherapy, including: Brain-wide functional architecture remodeling 1. substitutive strategies of harm reduction: substitution of by alcohol dependence and abstinence provides consumption modalities and/or substances and/or behaviours evidence for the three-stage hypothesis (e.g. how to cope diferently, starting from strategies applied Olivier George1, Daniel J Lurie2, Andres Collazo3, Max Kreifeldt1, by the patients and reinforcing what intuitively better works Harpreet Sidhu1, Mark D’Esposito2, Candice Contet1, Adam for them); 2. an integrative psychotherapy based on psycho- Kimbrough1 (1 La Jolla, USA, 2 Berkeley, USA, 3 Pasadena, USA) social rehabilitation modalities and introduced gradually, to frst address the most basic functions (life rhythms, negative Te identifcation of the psychological constructs and neuro- emotions, etc.) then the more and more complex issues biological mechanisms underlying the transition to addiction (social processes, cognitions); 3. and a behavioral pharmaco- remains one of the most critical steps to better understand logy according to medication’s mechanism of action (neuro- and treat alcohol and drug addiction. Converging lines of science-based nomenclature) allowing for treatment of ele- evidence suggest that multiple neurobiological modules mentary dimensions and endophenotypes, complemented by processing reward, incentive salience, habits, stress, pain, and specifc treatments when categorical diagnosis is possible. In executive function may explain the vulnerability to alcohol our experience, such an integrated and integrative approach addiction, and three major theories – incentive salience, allows for efcient treatment of the most complex patients in hedonic allostasis and habit – have been proposed to contri- an outpatient setting. bute to addiction. However, because of technical limitations, we have been unable to directly test these hypotheses and Mechanisms underlying binge drinking and visualize changes throughout the whole brain at single-cell compulsive alcohol use resolution in subjects that are dependent on alcohol to vali- date the existence of these modules. Te present study used an unbiased single-cell whole-brain imaging approach to map Heidi MB Lesscher (Utrecht, The Netherlands) neuronal activity in alcohol-dependent mice and found that alcohol abstinence resulted in whole-brain reorganization of Behavioural traits and neurobiological mechanisms functional architecture and a pronounced decrease in modu- underlying loss of control over alcohol use in rats larity not observed in moderate drinkers. Structuring of the Johanna AS Smeets1, A. Maryse Minnaard1, Geert MJ Ramakers1, alcohol abstinence network revealed that addiction is driven Roger AH Adan1, Louk JMJ Vanderschuren1, Heidi MB Lesscher1 by three major brain modules, reminiscent of the three-stage (1 Utrecht, The Netherlands) theory of addiction. Many hub brain regions controlling this pathological network were identifed, including several that Alcohol use disorder (AUD) is characterized by loss of have been typically overlooked in addiction research. Further, control over alcohol use, but only a minority of individuals a handful of the hub regions identifed were predictive of

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 269 Congress addiction-like behavior. Tese results provide a single-cell AUD. Baclofen may be particularly advantageous in those resolution map for addiction and demonstrate that alcohol with liver disease, due to its limited hepatic metabolism and dependence remodels brain-wide functional architecture to safe profle in this population. Baclofen is mostly used of- decrease modularity similarly to other brain disorders. Such label in some European countries and Australia, and in parti- neuroadaptations strongly reinforce the brain disease model cular, for patients who have not beneftted from the currently and may explain why addiction is such a pervasive disease and used and approved medications for AUD. In France, baclofen why relapse is so common. has been extensively studied and was recently approved at the dose of up to 80 mg per day, by the French authority that Face and predictive validities of a new preclinical regulates drugs approval and marketing (see Dr. Benjamin model of operant binge drinking Rolland’s talk). However, the use of this drug remains Jérôme Jeanblanc1, Pierre Sauton1, Maria del Carmen 1 1 1 controversial, in part due to uncertainty regarding dosing and Gonzalez-Marin , Alessia D’Ippolito , Virginie Jeanblanc , efcacy, alongside concerns about safety. A recent Consensus 1 1 Mickael Naassila ( Amiens, France) Statement among 26 international experts in the feld was developed and published (Agabio et al. Lancet Psychiatry Binge drinking has multiple defnitions in Humans (OMS, 2018) where the current state-of-the-art was briefy summa- NIAAA, binge score for research…) making it difcult to rized and the need for future research was emphasized. On defne it accurately in animal models. Here we developed the latter point, human laboratory studies may shed light an animal model in outbred rats (Long Evans males and on the biobehavioral and other mechanisms how baclofen females) based on an operant ethanol self-administration may work in some individuals with AUD (see Dr. Mehdi paradigm. Rats were trained to self-administer a 20% ethanol Farokhnia’s talk). Finally, beyond baclofen, positive allosteric solution under a FR1 (fxed ratio 1) then FR3 schedule for 1 modulation of the GABA-B receptor may represent a better hour. Slowly, the duration of the session was reduced frst to pharmacological approach towards the development of novel 30 minutes then to 15 minutes and the levels of consump- treatments for patients with alcohol and substance use disor- tion reached on average 1.2g/kg within 15 minutes sessions. ders (see Dr. Eric Augier’s talk). We found that such high consumption is associated with a higher motivation for the drug and for highly concentrated Biobehavioral mechanisms underlying baclofen’s alcohol solutions (30% vs. 10%). Moreover, we showed that effects on alcohol seeking and consumption: the speed of consumption is the factor that can diferentiate lessons learned from a human laboratory study heavy drinkers compared to binge drinkers. We also found Mehdi Farokhnia1, Sara Deschaine1, Armin Sadighi2, Melanie that prolonged binge drinking leads to a decrease in decision Schwandt1, Lisa Farinelli1, Mary Lee1, Fatemeh Akhlaghi2, making in a gambling task and that poor decision making is Lorenzo Leggio1 (1 Bethesda, USA, 2 Kingston, USA) associated to lower dopamine release in the nucleus accum- bens. We then tested drugs used in the treatment of alcohol Te GABA-B receptor agonist baclofen has been broadly use disorders (Acamprosate, (R)-Baclofen, GHB, Nalmefene studied and used as a pharmacotherapy for alcohol use disor- and Naltrexone) and we found that all of them reduced binge der. Te biobehavioral mechanisms underlying baclofen’s drinking and all of them but acamprosate decreased both efects are, however, not well understood. Human laboratory the motivation to consume and the relapse after prolonged studies provide an informative platform to shed light on this abstinence. We thus demonstrated the face and predictive domain. In the present randomized, double-blind, placebo- validities of our model and further neurobiological and controlled study, thirty-four alcohol-dependent individuals behavioral studies are in progress to better characterize this received baclofen (30 mg/d) or placebo for a week, and damaging behavior. then participated in a laboratory experiment consisting of three procedures: alcohol cue-reactivity, priming, and self- administration. Repeated blood samples were also collected Targeting GABAB receptors to treat AUD: recent for pharmacokinetic measurements. Group analyses showed advances in clinical and preclinical models that baclofen, compared to placebo, did not signifcantly attenuate cue-elicited craving or the amount of alcohol self- Eric Augier (Linköping, Sweden) administration. However, baclofen disrupted the link between alcohol priming and self-administration, as indicated by signifcant interaction efects between drug condition (baclo- The use of baclofen for patients with alcohol use fen vs. placebo) and some of the priming variables (alcohol disorder: where do we stand? craving: F3,9=6.03, p=0.01; alcohol sedation: F3,6=7.16, p=0.01; 1 1 2 Lorenzo Leggio , Mehdi Farokhnia , Roberta Agabio breath alcohol concentration: F1,25=5.22, p=0.03) on the total (1 Bethesda, USA, 2 Cagliari, Italy) amount of alcohol self-administered. Considerable interindi- vidual variability in baclofen pharmacokinetic parameters was Baclofen, a selective gamma-aminobutyric acid-B (GABA-B) observed. Maximum plasma concentrations of baclofen nega- receptor agonist, has emerged as a promising drug for Alcohol tively correlated with cue-induced alcohol craving (r=-0.57, use disorder (AUD). Tis talk will provide an overview of the p=0.03) and priming-induced ratings of ‘like more’ (r=-0.59, clinical work done with this medication in patients with p=0.02). Tese data suggest that baclofen may work by disso-

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 270 Congress ciating the link between an initial drink and subsequent alco- receptor PAM. Importantly, these efects are observed in hol consumption. Considerable pharmacokinetic variability the absence of signifcant sedative side efects. Finally, I will is an important factor to take into account when employing show new data evaluating the potential of positive allosteric baclofen as a treatment for alcohol use disorder. modulation of GABAB receptors to rescue pathological alco- hol choice over high value alternative rewards. France and the recent approval for baclofen in AUD Benjamin Rolland1 (1 Lyon, France) Neuroimaging in addiction: recent advances In October 2018, the French Drug Agency granted an appro- in the monitoring and prediction of val to the GABA-B receptor agonist baclofen for Alcohol pharmacological effects Use Disorder (AUD). Baclofen is thus now labeled for “sup- porting drinking reduction in AUD”, up to the dose of 80 mg Patrick Bach (Heidelberg, Germany) per day, and after failure of other drugs approved for AUD. Tis regulatory decision results from a long story of of- label use, sometimes at doses exceeding 300 mg per day. Te Effects of High-dose baclofen on neural and French practice consists of using baclofen in patients who are behavioural cue-reactivity in alcohol dependence still displaying heavy drinking. As baclofen is a sedative drug, Anne Beck1 (1 Charité, Berlin) interaction with alcohol can raise safety concerns. In 2019, many uncertainties remain with respect to the efcacy and Increased functional brain response towards alcohol-associated tolerability features of baclofen in AUD. Despite this, France stimuli (“cue reactivity”) is a neural hallmark of alcohol de- is now the frst country in which baclofen is ofcially labeled pendence and a promising target for pharmacotherapy. In this for AUD. Te French drug agency explained that this deci- study, we assessed the efects of individually titrated high- sion was not based only on scientifc considerations, but also dose baclofen on cue-induced brain activation in alcohol- on the pragmatic statement that more than 60,000 patients dependent (AD) patients in a randomized controlled trial were still treated with baclofen for AUD in France. As such (RCT). Patients receiving baclofen showed a significant the country will thus constitute an interesting real-life labo- stronger decrease in cue-elicited brain activation in left orbito- ratory regarding the public health impact of this medication frontal cortex (OFC), bilateral amygdala and left VTA than in AUD patients. patients receiving placebo and had signifcantly reduced

Effect of the novel GABAB PAM ADX74441 on relapse rates. Tus, our data suggest the modulatory capacity preclinical models of AUD and pathological alcohol of high-dose baclofen on alcohol-associated cue reactivity choice on a neuronal level, thereby potentially contributing to the Eric Augier1, Russell Dulman1, Gaëlle Augier1, Markus Heilig1 relapse preventive effects of this compound in alcohol (1 Linköping, Sweden) dependence. The ICCAM platform: using fMRI to characterize Alcohol effects on gamma-aminobutyric acid (GABA) brain responses in addiction and their transmission are key for the development and maintenance pharmacological modulation of alcohol addiction. Previous research indicate that GABAB Anne Lingford-Hughes1 (1 London, UK) receptor agonists such as baclofen can afect addiction-related behaviors in preclinical models of alcoholism. More impor- Tis talk will describe the ICCAM platform which uses 3 tantly, baclofen has also shown promise in clinical studies, in fMRI tasks to characterise brain responses in alcoholism particular in severely alcohol-dependent patients. However, and polydrug (alcohol, , ) addiction. Te study despite promising results in both clinical and preclinical mo- explored how any dysregulation is modulated by a range of dels, baclofen itself has inherent limitations as a therapeutic pharmacological probes, e.g. a DRD3 antagonist, a NK1 for alcohol addiction, and failed to obtain an approval for this antagonist, opiate antagonists and if this is consistent with indication. An attractive alternative approach to targeting the likely therapeutic beneft. same mechanism is ofered by positive allosteric modulators Comparison of the effects of naltrexone on cue (PAM:s) of the GABAB receptor, which have the poten- tial to achieve mechanistic and therapeutic efects similar reactivity across different substance use disorders to GABAB agonists, while avoiding tolerance and overdose Joar Guterstam (Stockholm, Sweden) toxicity. In this symposium, I will present recent data obtai- ned with ADX71441, a novel GABAB PAM that has entered Te naltrexone is often used in the treat- Phase 1 clinical testing, on several alcohol-related behaviors ment of alcohol and opioid use disorders, and some clinical in rats that model important aspects of human alcoholism. trials have also shown that it might reduce the risk of relapse In particular, ADX71441 dose-dependently decreased alcohol in amphetamine dependence. In recent years, a number of self-administration, with a higher efcacy in animals with a fMRI studies have investigated the efects of naltrexone on history of dependence. Furthermore, both cue- and stress- drug cue reactivity in individuals with these diferent subs- induced alcohol seeking were blocked by the GABAB tance use disorders. Several studies have reported that nalt-

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 271 Congress rexone attenuates neural responses to alcohol cues in alcohol From Fetal alcohol syndrome to Korsakoff dependent patients, and there is also preliminary evidence of syndrome through binge drinking: a a similar efect in patients with opioid use disorder. Recent neuroscientist/neuropsychological perspective studies of amphetamine users have found that they often exhibit strong neural and behavioral cue reactivity, but these Anne-Lise Pitel (Caen, France) reactions do not seem to be signifcantly afected by nalt- rexone pre-treatment. Tese divergent fndings might point to diferences in the pathophysiology of craving in alcohol, Evaluating and training executive functions in opioid and stimulant use disorders. binge drinking: a combined neuroscience approach Identifying neurobiological predictors for Pierre Maurage1, Valérie Dormal1, Séverine Lannoy1,2 (1 Louvain- pharmacological treatment response in addiction: la-Neuve, Belgium, 2 Stanford, USA) results from the recent TRANSALC study Patrick Bach1 (1 Mannheim, Germany) Binge drinking, constituting the most frequent alcohol consumption pattern among adolescents and young adults, Despite the high prevalence of alcohol use disorder (AUD), is characterized by a repeated alternation between intense only a few medications are approved for its treatment and consumption episodes and abstinence periods. Te neuro- meta-analyses point towards a modest overall efect size of cognitive defcits related to this drinking pattern have been available medications, such as Naltrexone. Understanding widely explored during the last decade. Executive functions the neural and behavioral mechanisms underlying the highly defcits have been specifcally identifed as key factors in the variable treatment response to anti-relapse medications emergence and maintenance of such habit. Tis talk will therefore seem to be a key factor for improving individual present new neuropsychological, electrophysiological and treatment success and enhancing impact on clinical practice neuromodulation data allowing to better understand the spe- based on the principles of precision medicine. We will present cifc executive impairments associated with binge drinking. data of a recent longitudinal open-label trial, investigating We will centrally underline that binge drinkers (1) present whether Naltrexone (NTX) could block increases in alcohol a diferential impairment across executive functions, with a craving and neural alcohol cue-reactivity (CR) in patients preserved performance for shifting and updating abilities, with alcohol use disorder, compared to standard treatment but impaired inhibition processes; (2) show a dissociation, using longitudinal combined neuroimaging and psychometric observed at the electrophysiological level, between impaired assessments. At baseline (before treatment initiation), all par- error-related processing and preserved feedback processing; ticipants underwent baseline psychometric testing and fMRI (3) have a sufciently preserved brain plasticity to beneft assessment of mesolimbic alcohol CR. Following this, pa- from neurostimulation-based rehabilitation of executive tients participated in a standard treatment program with the functions. Te theoretical, experimental and clinical impact of option of adjuvant NTX. After 2 weeks of treatment, AUD these new insights will then be discussed, notably to under- patients underwent a second combined neuropsychological line the potential usefulness of joint neuropsychological/ and fMRI assessment of alcohol craving and mesolimbic CR. neuromodulation interventions among people presenting Results show higher mesolimbic CR in AUD patients vs. binge drinking habits. healthy controls at baseline. Over the treatment episode of 2 Why we should ask bingers if they smoke weeks, mesolimbic CR signifcantly increased in the standard cannabis? treatment group (n=13), but not in the NTX group (n=22, Hélène Beaunieux1, Virginie Bagneux1, Ludivine Ritz1, Ingrid F(1,12)=23.526, p=0.001). Only NTX treated patients showed Banovic2, Anaelle Bazire1, Nicolas Cabé1, Caroline Cheam- significant attenuation of CR in the left putamen over Bernière1, Laure Marine Houel1, Denis Jacquet1, Reynald time (interaction time x medication: F(1,33)=6.823, p=0.013) 1 1 1 that was associated with a reduced relapse risk to heavy- Le Boisselier , Pascale Leconte , Jean-Baptiste Marchand , 1 1 2 drinking within three months of treatment (interaction Nicolas Margas , Maxime Mauduy , Fabrizio Scrima , Cécile 1 1 1 2 treatment x time: Hazard Ratio=0.255, 95%CI=0.084-0.775, Sénémeaud , Jessica Mange ( Caen, France, Rouen, France) p=0.016). Further, NTX treated patients compared to patients receiving standard treatment reported a signifcantly higher Studies focusing on college students’ consumptions of psycho- proportion of abstinent days during follow-up (t(33)=1.834, active substance and their consequences have mainly focused p=0.042). In conclusion, NTX blocked increased in mesolim- on alcohol use and more recently on binge drinking (BD). bic CR that was observed in the standard treatment group. Distinct BD patterns associated with specifc psychological NTX was most efective in the patients with high baseline profles have been identifed (Lannoy et al., 2017; Gierski CR in the left putamen, refecting in a number needed to et al., 2018). Based on its efects on various psychological treat of 1.8 [95%CI=1.3-6.2] to prevent one heavy relapse. parameters, cannabis use, which frequently co-occurs with While the results from our naturalistic study await further BD, may modulate these diferentiated psychological profles. confrmation from prospective randomized trials, they sup- Further, in contrast to the wide evidence of neuropsycholo- port the role of neural CR as a biomarker in the development gical defcits associated with the use of cannabis or alcohol of precision medicine approaches with NTX. separately, few studies have investigated the risk of alcohol

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 272 Congress use disorder and neuropsychological defcits related to the such a severe and debilitating neurological disease in order combined consumption of alcohol and cannabis. Te aims to provide appropriate prevention treatment. of the present study were to examine the efect of cannabis on (1) the risk for alcohol use disorder and (2) the neuro- psychological defcits observed in bingers. First, students of Genetics/genomics in alcohol induced liver the University of Caen consuming alcohol and/or cannabis injury – what next? were screened through an internet survey-based study focu- sing on alcohol and cannabis experiences. Results showed Vanessa Rausch (Sydney, Australia) that compared to bingers, students who both binged and smoked cannabis had an earlier onset of alcohol consump- tion, drank more and were more at risk of alcohol use disor- Genetics of hepatic steatosis and fibrosis der. Motivation and socio-normative parameters associated Marcin Krawczyk1 (1 Homburg, Germany) with alcohol consumption were signifcantly higher in this group than in binge drinkers who did not smoke cannabis. Fatty liver disease (FLD) belongs to the most frequent Te neuropsychological evaluation revealed that compared to conditions in hepatology. Indeed, more than 20% of adult bingers, students who binged and smoked cannabis had more Europeans sufer from fatty liver and the incidence of this severe neuropsychological impairments, especially for episo- condition is predicted to increase even further. A subgroup dic memory. Tese results suggest that cannabis consumption of patients with FLD will also develop liver fbrosis, which associated with BD is a risk factor for alcohol use disorder is a common hallmark of chronic liver diseases. Both hepatic and episodic memory defcits. Tose fndings reinforce the lipid accumulation as well as liver scarring have for long been idea that BD prevention programs may gain efcacy if consi- expected to be modulated by the inherited predisposition. In dering its frequent combination with cannabis. the recent years genetic variants in several genes, for example Decorticating the pathophysiological mechanisms PNPLA3, TM6SF2 and MBOAT7, have been linked to the progression of chronic liver diseases. Prosteatotic and/or underlying alcohol use disorder with and without profbrotic variants in these genes were frst detected in large Korsakoff’s syndrome: a neuroimaging review and genomeh wide association studies (GWAS) and afterwards future directions 1 1 1 these associations were replicated in the following candidate Shailendra Segobin , Alice Laniepce , Nicolas Cabé , François gene analyses. In particular carriers of the PNPLA3 p.I148M 1 1 1 1 Vabret , Francis Eustache , Anne-Lise Pitel ( Caen, France) variant have been proven to be at risk of severe liver steato- sis, fbrosis, cirrhosis and hepatocellular carcinoma (HCC) Alcohol Use Disorder (AUD) exists in two main clinical rendering variant PNPLA3 a common genetic risk factor forms. Te frst one, often referred to as “uncomplicated for progressive liver injury. Interestingly, the same variant AUD”, is associated with mild-to-moderate defcits of epi- also seems to modulate the response to the FLDh therapies. sodic memory, executive functions, working memory and Te most recently detected splice variant rs72613567 in motor abilities. Some, but not all, uncomplicated AUD hydroxysteroid 17h dehydrogenase 13 (HSD17B13) seems patients develop the second clinical form that is Korsakof ’s to, in turn, reduce the risk of FLD. Here we will summarize syndrome, characterized by severe and irreversible amnesia, the current knowledge on the genetic background of hepatic resulting from long-term excessive alcohol consumption steatosis and fbrosis, discuss the efects of the known variants and thiamine defciency. Clinically, it is extremely relevant on the disease progression as well address the potential use to identify uncomplicated AUD patients at risk of deve- of genetic analyses in the clinical workh up of patients with loping Korsakof ’s syndrome. In this talk, the contribution FLD. of neuroimaging biomarkers towards understanding the pathophysiological mechanisms underlying these two clini- An exploratory genomep wide analysis of patients cal forms will be reviewed. More precisely, the brain circuits with alcoholic hepatitis predominantly afected in both clinical forms, namely the Suthat Liangpunsakul1 (1 Bloomington, USA) frontocerebellar and Papez circuits will be discussed in terms of alterations to their macrostructural and microstructural An exploratory genomeh wide association study (GWAS) integrity. Te thalamus, a key region consisting of several was conducted comparing patients with alcoholic hepatitis nuclei is shared by these two brain circuits. Consolidating (AH) and heavy drinking matched controls without liver evidence showing that the loci and nature of structural disease in order to identify variants or genes associated with alterations occurring within the thalamus potentially defnes risk for AH. Individuals were genotyped using the multih the specifcity of Korsakof ’s syndrome will be presented. To ethnic genotyping array, after which the data underwent conclude this talk, current and prospective biomarkers in the conventional quality control. Using bioinformatics tools, feld of cognition, neuroimaging and biology will be discussed pathways associated with AH were explored on the basis of based on how they bring incremental value towards the global individual variants, and based on genes with a higher ‘burden’ understanding of the pathological mechanisms underlying of functional variation. Although no single variant reached Korsakoff’s syndrome. Such a better understanding will genomeh wide signifcance, an association signal was obser- ultimately enable clinicians to identify patients at risk for ved for PNPLA3 rs738409 (p=0.01, OR 1.9, 95%CI 1.1h

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 273 Congress

3.1), a common single nucleotide polymorphism that has investigate the impact of these variants on important clinical been associated with a variety of liverh related pathologies parameters in response to alcohol withdrawal. Terefore, including alcoholic cirrhosis. Using the improved gene set we prospectively enrolled 516 ALD patients for alcohol enrichment analysis for GWAS tool, it was shown that, based detoxifcation. Patients were genotyped and CAP, LS and on the single variants’ traith association ph values, multiple ultrasound as well as laboratory markers were assessed before pathways were associated with risk for AH with high conf- and after detoxifcation. In 105 patients, liver biopsy was also dence (false discovery rate [FDR]<0.05), including several obtained and histologically analyzed. Carriers of MBOAT7 pathways involved in lymphocyte activation and chemokine CC and PNPLA3 GG showed a strong and combined efect signaling, which coincides with fndings from other research on fbrosis development (P<0.05), however display striking groups. Several Tox Functions and Canonical Pathways were diferences with regard to infammation, fbrosis and steato- highlighted using Ingenuity Pathway Analysis, with an espe- sis in response to alcohol withdrawal. Infammation was not cially conspicuous role for pathways related to ethanol degra- diferent and resolved equally in all MBOAT7 genotypes dation, which is not surprising considering the phenotype of during detox (AST, P<0.001), whereas PNPLA3 GG car- the genotyped individuals.Tis preliminary analysis suggests riers presented with signifcantly enhanced liver injury after a role for PNPLA3 variation and several gene sets/pathways detoxifcation and with delayed resolution of infammation that may infuence risk for AH among heavy drinkers. (M30 and AST, P<0.001). Finally, steatosis resolved equally in both polymorphisms and genotypes to the same extent and Contribution of common SNPs in explaining no diferences prior and after detoxifcation have been obser- genetic variability in alcoholic cirrhosis ved. In the histology cohort, PNPLA3 GG was signifcantly 1 1 Devanshi Seth ( Sydney, Australia) associated with infammation (steatohepatitis, ballooning and lobular infammation) and steatosis and MBOAT7 CC with Genetic pathways contributing to the pathophysiology of fbrosis. In summary, both variants are associated with fbrosis liver cirrhosis in drinkers are fundamentally important to un- progression, but the response of steatosis and liver injury to derstand this disease. Tere is limited comprehension of the alcohol withdrawal is remarkably diferent. While PNPLA3 genetic basis of variation in Alcoholic Liver Cirrhosis (ALC) is associated with liver injury and steatosis, MBOAT7 is not susceptibility as only up to 20% of heavy chronic drinkers afecting liver injury. Interestingly, both variants did not alter progress to cirrhosis. Cirrhosis is the major medical conse- the resolution of steatosis during alcohol withdrawal. quence and health pro blem of excessive alcohol abuse with Alcoholic liver cirrhosis, beyond single variant high morbidity and mortality. Genetics of ALC is poorly understood despite several candidate gene studies and a single analysis 1 1 GWAS reporting a strong association with PNPLA3. Other Tae-Hwi Linus Schwantes-An ( Bloomington, USA) reported SNPs, e.g. TM6SF2, MBOAT7 and HSD17B13, only show modest GWAS level association with ALC. Our In the posth GWAS era, much of the genetic underpinnings multin ational GenomALC Consortium also performed of complex diseases such as alcoholic liver cirrhosis (ALC) GWAS in the world’s largest collection of drinkers in a caseh remain unexplained. Missing heritability, the large unex- control study design. Age, gender and ethnicity matched plained portion of genetic heritability after discoveries from Cases (drinkers with cirrhosis) and Controls (drinkers with GWAS studies, spurred adaptation of nexth gen sequencing no liver disease) were subjected to GWAS (Infnium GSA to identify genetic variations that are not captured by GWAS Array). Our data showed increased risk of alcoholic cirrhosis arrays. In parallel, statistical genetic methods have evolved in ofsprings of parents with alcohol problems who died of from simple single variant analysis (e.g. assessing efect of liver disease, underscoring the heritability of this disease. We each genetic variant one at a time) to polygenic risk scores confrmed PNPLA3 (rs739409) and HSD17B13 (rs4607179) that include more than several tens of thousands of common that strongly associated with alcoholic cirrhosis. However, all variants to stratify risk for disease. In this talk, using examples these common variants identifed from GWAS approach only from GenomAlc Consortium analyses, examples of recent account for about 20% of the overall genetic variance leaving statistical genetic methods will be highlighted. much of the genetic contribution to ALC unexplained. Due to typical GWAS array design, which focuses on common variants, protein coding exonic and rare variants, as well as The role of Cytochrome P4502E1 in alcoholic other non-genetic factors are yet to be for ALC. liver disease and cancer The role of PNPLA3 and MBOAT7 polymorphisms Helmut K Seitz (Heidelberg, Germany) during alcohol detoxification – identification of different mechanisms for fibrosis development Vanessa Rausch1 (1 Heidelberg, Germany) Ethanol metabolism in the liver: the role of CYP2E1 S. Zakhari1 (1 Washington, USA) Te PNPLA3 rs738409 and MBOAT7 rs626283 polymor- phisms are genetic risk factors for ALD progression; however, About 95-98% of ingested alcohol is metabolized in the liver their molecular mechanisms are still poorly understood. We in a two-stage, enzymatically-catalyzed oxidation process;

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 274 Congress the remainder is excreted in breath, urine and sweat. A small vation of carcinogens and potential tumor development in the proportion of alcohol metabolism occurs via non-oxidative gastrointestinal tract of patients with an alcohol problem. In metabolic pathways resulting in the formation of fatty acid essence, microsomal CYP2E1 plays an essential role in drug- ethyl esters and phosphatidyl ethanol. A smaller portion alcohol interactions, increased risk of toxicity in the liver, and undergoes conjugation with glucuronic acid or sulphate, and potential tumor development in the gastrointestinal tract. these conjugates are excreted in urine. Te major pathway of References oxidative metabolism of ethanol in the liver involves cytosolic - Teschke, R. Biomedicines 2018; 6, 106. alcohol dehydrogenase (ADH) to produce acetaldehyde. Te - Teschke, R. Alcoholism, Clinical and Experimental Research 2019; 43: 386- cytochrome P450 isozymes, including CYP2E1, 1A2 and 400. 3A4, also contribute to ethanol oxidation to acetaldehyde in the liver, particularly at elevated alcohol concentrations. CYP2E1, ethanol and carcinogenesis 1 1 CYP2E1 is induced by chronic ethanol consumption. It also Gary J Murray ( Bethesda, USAh metabolizes numerous medications such as acetaminophen, and other xenobiotics. Te catalase enzyme can also metabo- Although there is a strong association between chronic and lize alcohol to acetaldehyde; however, this pathway appears excessive consumption of alcohol and increased risk of va- to play a minor role in alcohol oxidation by the liver. Tis rious cancers, the causative role and the specifc mechanisms presentation focuses only on CYP2E1 and will address its involved remain a subject of ongoing research. Cytochrome role in: health and disease; ethanol-mediated oxidative stress; P4502E1 (CYP2E1) is induced in the liver after chronic al- drugs, xenobiotics and procarcinogens metabolism; fatty acid cohol consumption. As a consequence, in addition to alcohol, metabolism; and ethanol-induced hepatotoxicity and carci- there is increased oxidation of many toxic and carcinogenic nogenesis. Discussion will also focus on hepatic CYP2E1 in xenobiotics, including a variety of drugs, steroids, and other pathological conditions such as obesity, diabetes and chemical compounds. Te oxidation and reduction reactions catalyzed inducers, as well as on drugs that inhibit ethanol-induced by most P450 class enzymes are important mechanisms for CYP2E1 and their role as protective agents against ethanol- detoxifcation of these xenobiotics but these same processes mediated liver injury. may lead to the creation of damaging products including the direct activation of carcinogens by oxidation of less toxic pre- Alcohol and drug interaction: the role of CYP2E1 cursors. Another important pathway involves the generation 1 1 Rolf Teschke ( Frankfurt/Main, Germany) of reactive oxygen species (ROS) including lipid peroxidation products and 4-hydroxynonenal that react with DNA resul- Following the discovery of the hepatic microsomal ethanol- ting in the formation of exocyclic etheno DNA-adducts. Tese oxidizing system (MEOS) by Charles S Lieber and Leonore highly carcinogenic adducts contribute to the development M DeCarli in 1968 and its subsequent purifcation and of more serious pathology in liver and other tissues. Tere isolation from alcohol dehydrogenase and catalase through remains some controversy on the specifc role of CYP2E1 column chromatography in 1972, additional studies identifed in the production of ROS and of oxidative damage, and the the microsomal cytochrome P450 (CYP) with its isoenzyme signifcance of the observations that microsomes and purifed CYP2E1 as its major constituent. CYP2E1 metabolizes P450s generate ROS, has been questioned. Tis is countered not only ethanol but also other short chain alcohols and by observations in liver biopsies from patients with alcoholic various drugs and chemicals. Among these are paracetamol, liver disease (ALD) that the generation of these adducts were halothane, and carbon tetrachloride. Consequently, the broad correlated with the induction of CYP2E1 in the liver after substrate specifcity explains molecular interactions at the chronic alcohol consumption. Newer data has also emerged level of CYP2E1. In particular, a few substrates such as to implicate mitochondrial CYP2E1 in the production of disulfram, diallylsulfde, and clomethiazole are known for ROS. Te causative role of increased CYP2E1 activity in their inhibitory efect on CYP2E, whereas the use of many the development of alcohol-related cancers will be discussed. other chemicals and drugs including acetone and isoniazid upregulate CYP2E1 gene expression. Most importantly, The role of CYP2E1 in alcoholic liver disease and prolonged ethanol consumption upregulates CYP2E1 and alcohol mediated carcinogenesis thereby induces MEOS activity through a process involving Helmut K Seitz1, Sebastian Mueller1 (1 Heidelberg, Germany) reduced degradation of CYP2E1 by inhibition of hepatic proteasome peptidase activities. Tis induction of MEOS Various factors are involved in the pathogenesis of alcoholic activity explains the adaptive increase of alcohol metabolism liver disease (ALD) and ethanol mediated carcinogenesis. In in individuals with prolonged alcohol abuse. In addition, addition to genetic, epigenetic and immunologic mechanisms, upregulation of CYP2E1 and associated production of toxic acetaldehyde associated toxicity, oxidative stress as well as intermediates is responsible for increased acute liver injury cytokine mediated infammation are of major importance. by paracetamol or carbon tetrachloride in patients with a Oxidative stress with the generation of reactive oxygen spe- past history of alcohol abuse. Ethanol-related upregulation cies (ROS) develops either in infammation (alcoholic hepa- of CYP2E1 is also observed in the intestinal tract, modifying titis) or during oxidation of ethanol via cytochrome P4502E1 thereby the intestinal microbiome, considered as mechanistic (CYP2E1). CYP2E1 is induced by ethanol, oxidizes ethanol contributor to alcoholic liver injury, and facilitating the acti- to acetaldehyde and generates ROS during this process. ROS

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 275 Congress results in protein damage, enhanced fbrogenesis and DNA laboratory. However, GET73 appeared to reduce naturalistic lesions. Furthermore, CYP2E1 induction results in an en- alcohol consumption during the outpatient washout period. hanced activation of various procarcinogens and an increased Probenecid reduces alcohol drinking in rats: is degradation of retinol and retinoic acid (RA), a compound pannexin1 a novel therapeutic target for responsible for cell diferentiation and proliferation. An inhi- alcoholism? bition of CYP2E1 results in an improvement of ALD and 1 2 2 chemically induced carcinogenesis in animal experiments. In Pietro Paolo Sanna , Brendan J Tunstall , Sam A McConnell , 2 2,3 4 man, CYP2E1 is induced following the consumption of 40 Katrina L Gazo , Lia J Zallar , Carolina Haass-Koffler , Vez 1 2 2 grams of ethanol per day already after one week. However, Repunte-Canonigo , George F Koob , Leandro F Vendruscolo 1 2 3 4 the induction varies interindividually. Te mechanism for this ( La Jolla, USA, Baltimore, USA, Bethesda, USA, Providence, is still unclear. Patients with ALD show a signifcant correla- USA) tion between CYP2E1, the occurrence of highly carcinogenic etheno DNA-adducts and the severity of fbrosis. First results Te development of novel and more efective medications of the efect of CYP2E1 inhibition by chlormethiazole, a for alcohol use disorder (AUD) is a pressing unmet medical specifc CYP2E1 inhibitor on ALD can be expected soon. need. Approved medications for AUD generally have limited efcacy and are prescribed for fewer than 10% of US patients with AUD. Drug repositioning or repurposing is an appealing Drugs with novel mechanisms for the treatment strategy to bring new therapies to the clinic because it greatly of alcohol dependence reduces the overall costs of drug development and expedites the availability of treatments to those who need them. We Robert Swift (Providence, USA) recently found that the glycyrrhetinic acid derivative car- benoxolone (CBX; 3-hydroxy-11-oxoolean-12-en-30-oic acid 3-hemisuccinate), a medication that was previously ap- Effect of the mGluR5 modulator GET 73 on alcohol proved for the treatment of gastritis and peptic ulcer, reduces and pharmacodynamics and both dependent and nondependent alcohol intake in rodents, alcohol craving and consumption in a human suggesting that it is a candidate for drug repositioning for laboratory model the treatment of AUD. Carbenoxolone is a multi-target drug that shapes cellular responses to glucocorticoids by inhibiting Robert Swift1, Carolina Haas-Koffler1,2, Lorenzo Leggio2, 11-hydroxysteroid dehydrogenase (11-HSD) isozymes. Roberto Cacciaglia3 (1 Providence, USA, 2 Bethesda, USA, 3 San It also inhibits pannexin1 channels, which contribute to Remo, Italy) adenosine triphosphate release, in the extracellular space. Probenecid is a medication that is used clinically primarily GET 73 is a novel, small molecule compound that reduces to increase uric acid excretion in the urine in hyperuricemic alcohol consumption and has anxiolytic and anti-stress conditions, such as gout, through its activity as a competitive activity in animals. GET 73 acts as a negative allosteric substrate for anion-transporting polypeptides in the kidney. modulator (NAM) at the mGluR5 receptor and could Probenecid was also shown to inhibit pannexin1 channels. reduce the high glutamatergic allostatic state associated Terefore, we tested its efects on alcohol intake in rats. with alcohol use disorders. To establish safety/tolerability Similar to CBX, probenecid reduced alcohol intake in both and to determine whether GET 73 reduces alcohol craving dependent and nondependent rats. Tese results raise the and alcohol drinking, we conducted a placebo-controlled, possibility that pannexin1 may be a novel therapeutic target within-subjects crossover study with GET 73 in twenty non- for the treatment of AUD. Te clinical use of probenecid has treatment seeking alcohol dependent persons. After screening been found to be generally safe, suggesting that it may be a for medical and psychiatric suitability, eligible subjects were candidate for drug repositioning for the treatment of AUD. randomized to the 14-day inpatient study, receiving 3 days ANS-6637, a selective reversible inhibitor of ALDH2, of treatment with GET 73 or placebo, followed by a 7-day outpatient washout, followed by 3 days of the alternate medi- suppresses alcohol consumption and cue-induced cation. Under each condition (GET 73 or placebo), on Day alcohol self administration in the absence of 2 and Day 12, participants received an oral dose of alcohol alcohol and acetaldehyde 1 2 3 to bring BAC to 0.12 g/L. Alcohol and GET 73 pharma- Ivan Diamond , Stephanie O’Malley , Maria Arolfo, Lina Yao , cokinetics and pharmacodynamics (intoxication, impairment, Peidong Fan3, Brent Blackburn1 (1 Los Angeles, USA, 2 New mood, etc.) were monitored and compared between drug Haven, USA, 3 Redwood City, USA) and placebo conditions. On Day 3 and Day 13, participants received an alcohol cue-reactivity (craving) session, followed According to prevailing clinical concepts Disulfram dis- by alcohol self-administration. Te results showed that GET courages drinking because of adverse symptoms caused 73, was safe and did not afect alcohol pharmacokinetics. by increased acetaldehyde as a consequence of irreversible GET 73, compared to placebo, increased alcohol sedation on inhibition of hepatic ALDH2. However, Daidzin, a known the BAES but did not afect performance. GET73 did not ALDH2 inhibitor derived from Kudzu extracts, suppresses afect subjective craving or alcohol self-administration in the Golden Syrian hamster drinking without increasing ace-

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 276 Congress taldehyde. Tis suggested that inhibiting ALDH2 in brain with AUD. Tis research was supported by U01AA025476 might reduce urges to drink alcohol. We used highly selec- and P60AA006420. tive reversible inhibitors of ALDH2 to prevent excessive self-administration of alcohol. Paradoxically, these inhibitors also inhibited cue-induced reinstatement of drinking in the Recent advances in probing the link between absence of alcohol and acetaldehyde. We soon discovered that alcohol and neuroendocrine pathways ALDH2 inhibition in VTA prevents dopamine surges under- lying craving/drug-seeking for alcohol, cocaine and other Carolina L Haass-Koffler (Providence, USA) addictive drugs. Here we demonstrate that expression of viral ALDH2 antisense in the VTA also suppresses drinking, in- dependently confrming ALDH2 as a CNS target. Transient The effects of oral and intravenous alcohol ALDH2 antisense expression correlates directly with tran- administration on appetitive and stress-related sient reduction of alcohol drinking. We then developed ANS- hormones: results from human laboratory 6637, a safe highly selective reversible inhibitor of ALDH2. experiments We studied the dose-response of ANS-6637 interaction Mehdi Farokhnia1 (1 Bethesda, USA) with alcohol in subjects consuming 5 standard drinks in 2.5 hours. Tere were virtually no adverse efects from ANS-6637 A growing body of evidence from preclinical and clinical except an insensitive fushing reaction. We know ANS-6637 research suggest that endocrine pathways play important only targets ALDH2. In contrast, Disulfram has recognized roles in the pathophysiology of addictive behaviors, including broad toxicity. Te adverse alcohol-dependent adverse efects alcohol use disorder (AUD). A number of these pathways, es- of Disulfram are likely due to strong inhibition of hepatic pecially appetitive and stress-related hormones, are currently ALDH1, not merely ALDH2. Our fndings suggest that under investigation as potential targets to develop novel treat- ANS-6627 holds great promise for treating alcoholism and ments for AUD. To this end, it is also important to under- preventing relapse by attenuating craving/alcohol seeking stand whether and how alcohol consumption and dependence behavior without immediate adverse efects. may afect endogenous concentrations of endocrine markers. An evaluation of the Peroxisome proliferator In a series of human laboratory experiments, we examined the efect of alcohol administration on blood concentrations receptor-alpha (PPAR-) agonist, fenofibrate, in a of diferent endocrine markers in heavy-drinking alcohol- human laboratory model of alcohol use disorder dependent individuals. Four separate sessions were conducted 1 1 Barbara J Mason ( La Jolla, USA) across these studies: oral self-administered (variable dose) alcohol, oral fxed dose alcohol, intravenous self-administered Te PPAR- agonist, fenofbrate, has been shown to decrease (variable dose) alcohol, and intravenous fxed-dose alcohol. alcohol consumption and preference in rats and mice, and a Repeated blood samples were obtained during each session human genome-wide association study showed an association and the following hormones were measured: total ghrelin, of a single nucleotide polymorphism in PPAR- with alcohol acyl-ghrelin, leptin, -like peptide 1, GLP-1, insulin, withdrawal. Taken together, these data provide a compelling , (PP), peptide YY, gastric inhi- rationale for testing fenofbrate for therapeutic potential in bitory peptide (GIP), insulin-like growth factor 1 (IGF1), a human laboratory model of AUD. We hypothesized that (GH), prolactin, Adrenocorticotropic fenofbrate would signifcantly attenuate craving in response hormone (ACTH), cortisol, and aldosterone. Te results of to in vivo alcohol cue exposure in the laboratory and reduce alcohol administration, via diferent routes and with diferent drinking under natural conditions during treatment and dosing schedules, on each endocrine marker will be discussed. post-treatment follow-up. Fifty non-treatment-seeking, cue- reactive volunteers with AUD (39 males, 11 females; aged Dysregulations of glucocorticoid and 37.6 ± 11.8 years) were randomized to 9 days of treatment mineralocorticoid receptor systems in alcohol with fenofbrate (145mg/d) or matched placebo, and were dependence: converging evidence from rats and followed for 1-week post-treatment. Subjects were required humans to be abstinent for 3 consecutive days prior to testing on Day Leandro F Vendruscolo1 (1 Baltimore, USA) 9; abstinence was verifed by alcohol glucuronide testing. On Day 9, subjects were exposed to standardized mood induc- Alcohol consumption and withdrawal in alcohol use disorder tion procedures and in vivo beverage cues (alcohol or water). (AUD) activate the hypothalamic-pituitary-adrenal (HPA) Subjects consistently showed signifcantly greater craving axis to release corticosteroids (corticosterone in rats or cortisol in response to alcohol cues relative to water cues, but no in humans), which binds to glucocorticoid receptors (GRs) diferences in craving between fenofbrate and placebo were and mineralocorticoid receptors (MRs). We hypothesized that observed. Similarly, no pre-post treatment diferences were excessive activation of the HPA axis by alcohol intoxication found for drinking. Fenofbrate concentration in plasma did and withdrawal dysregulates GR and MR systems and that not correlate signifcantly with alcohol cue reactivity or drin- these changes contribute to negative emotional states that king measures. Tese data do not show an advantage in the- drive compulsive alcohol drinking. We found that GR expres- rapeutic potential for fenofbrate over placebo in individuals sion and function were altered in cortical and subcortical brain

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 277 Congress regions in dependent rats compared with nondependent rats. Efects of ghrelin administration on cortisol and aldoste- Systemic and intracerebral (central nucleus of the amyg- rone concentrations have been observed in ghrelin-exposed dala, ventral tegmental area, and nucleus accumbens) GR tissues or cells, rodents and healthy volunteers, however antagonism blocked the enhanced alcohol drinking and the whether these efects replicate in individuals with AUD is motivation for alcohol in alcohol dependent rats. In a trans- unknown. Here we tested the hypothesis that intravenous lational proof-of-concept human laboratory study, we found administration of ghrelin leads to increase in endogenous that GR antagonism decreased cue-induced alcohol craving serum cortisol and aldosterone concentrations in alcohol- and alcohol drinking in humans with alcohol use disorders. dependent heavy drinking individuals, and that these changes Additionally, we found that levels of MR in the CeA were may predict ghrelin-induced alcohol craving. This was a negatively associated with anxiety-like behavior and compul- double-blind, placebo-controlled human laboratory study in sive-like alcohol drinking in dependent rats. In non-abstinent non-treatment-seeking, heavy-drinking alcohol-dependent patients with AUD, the levels of aldosterone, an MR agonist, individuals randomized to receive either placebo, 1 mcg/kg positively correlated with alcohol drinking, craving and anxiety or 3 mcg/kg of intravenous ghrelin. Ten, participants unde- scores, suggesting a potential functional role of MR in AUD. rwent a cue-reactivity procedure in a bar-like setting, which Tese fndings provide converging evidence from rats and included exposure to both neutral (juice) and alcohol cues. humans for dysregulations of GR and MR systems in AUD. Repeated blood samples were collected and used to measure endogenous cortisol and aldosterone serum concentrations in Effect of GLP-1 analogue and desacyl ghrelin response to exogenous ghrelin administration. Furthermore, administration on alcohol cue reactivity in cortisol and aldosterone serum concentrations were used to humans: the gut hormone in addiction study develop a model to predict the efect of exogenous ghre- 1 1 Tony Goldstone ( London, UK) lin administration on alcohol craving. Intravenous ghrelin administration increased endogenous cortisol and aldosterone Common neurobiological mechanisms underlie addictive serum concentrations. While the efects on cortisol were behaviours, including alcohol use disorder (AUD), drug greater than those on aldosterone, only the ghrelin-induced dependence and overeating. Recent pre-clinical research changes in aldosterone serum concentrations predicted alco- has shown that gut-brain hormonal signals regulating food hol craving. Tese fndings provide evidence of ghrelin efects intake play an important role in non-food reward behaviours, on glucocorticoids and mineralocorticoids in individuals and that the role for the appetitive hormones, glucagon with AUD, thereby providing additional information on the like peptide-1 (GLP-1), and acyl ghrelin (AG), extends potential mechanisms how the ghrelin system may play a role beyond food intake regulation to include reward behaviour in alcohol craving and seeking in AUD. Funding: Dr. Haass- and consumption of alcohol. Although desacyl ghrelin, the Kofer is supported by the National Institute on Alcohol precursor for AG (active at the GHSR1a receptor), is not Abuse and Alcoholism (K01AA023867; PI: Haass-Kofer). an antagonist or inverse agonist at GHSR1a, it has opposite Drs. Farokhnia and Leggio are supported by the National metabolic efects to AG in some pre-clinical and clinical Institute on Alcohol Abuse and Alcoholism Division of studies, and reduced sugar intake in humans. Furthermore, Intramural Clinical and Biological Research and the National in clinical studies a DAG analogue, AZP-531, reduces body Institute on Drug Abuse Intramural Research Program (ZIA- weight and improves glycaemic control in adults with obe- AA000218, Section on Clinical Psychoneuroendocrinology sity and type 2 diabetes mellitus. Dr. Goldstone will present and Neuropsychopharmacology; PI: Leggio). Te parent study novel results from his MRC-funded experimental medicine, was funded by the National Institute on Alcohol Abuse and Gut Hormone in Addiction study, examining the efects of Alcoholism (R21AA019709; PIs: Leggio and Kenna). Te acute infusion of the GLP-1 analogue, Exendin-4, and DAG content of this article is solely the responsibility of the authors on brain responses to evaluation of alcohol pictures using and does not necessarily represent the ofcial views of the functional magnetic resonance imaging in 3 groups: adults National Institutes of Health or the Department of Veterans with obesity, ex-smokers and abstinent alcohol dependence Afairs. Confict of interest: the authors report no biomedical (http://clinicaltrials.gov/ct2/show/NCT02690987). This fnancial interests or potential conficts of interest. will reveal the possible benefts of targeting the GLP-1 and ghrelin systems for treatment of alcohol use disorder, and potential underlying mechanisms for reductions in alcohol Free oral communications consumption by attenuating alcohol cue reactivity. Intravenous administration of ghrelin increases In vivo longitudinal study of risky alcohol serum cortisol and aldosterone concentrations in consumption effects on the brain heavy-drinking alcohol-dependent individuals A. Lanquetin1, A. Drieu1, C. Freyssainge1, D. Vivien1, A.L. Pitel1, Carolina L Haass-Koffler1, Mehdi Farokhnia2, Lorenzo Leggio1 M. Rubio1 (1 Caen, France) (1 Providence, USA, Bethesda, USA) Introduction: neuroimaging and neuropsychological studies Increasing evidence supports the role of appetite-regulating revealed structural and functional brain alterations associated hormones, including ghrelin, in alcohol use disorder (AUD). with Alcohol Use Disorder (AUD). In 50 to 80 % of AUD

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 278 Congress patients, these brain alterations result in cognitive and/or the detection of KS patients and its relationships with brain motor impairments. However, the consequences of risky damage remain unknown. Ten KS patients, 26 AUD patients alcohol consumption (not reaching AUD but superior to the and 16 healthy controls (HC) underwent the BEARNI and recommendations) have been less studied in both humans a 3T-MRI examination. On BEARNI, KS had lower per- and rodents. In this longitudinal study, we aimed at studying formance than AUD patients (who did not difer from HC) the efects of risky alcohol consumption in mice at diferent for the episodic memory and fuency scores. Te specifcity of time points. Materials and methods: mice were divided into KS defcits on the memory subtests suggests that BEARNI 3 groups: control (drinking water), risky alcohol consump- is sensitive to amnesia. Statistical cluster analysis revealed that tion (10% ethanol solution ad libitum) and risky alcohol several AUD patients were classifed within the same cluster consumption with repeated periods of abstinence (ethanol as KS patients based on the BEARNI episodic memory replaced by water before and during the test week). From 6 results. Tus, a low score on this subtest (inferior or equal to weeks to 12 months alcohol exposure, every 3 months, and we 1.5 points/6) would enable the detection of patients at risk conducted i) a battery of behavioral tests to measure motor for developing KS. Multiple regression analyses conducted abilities (balance beam), anxiety (open feld) and memory between GM volume and performance on each BEARNI (Y-maze, fear conditioning); ii) MRI examinations to study subtest revealed correlations with the FCC, the PC and parie- regional brain volumes. Beverage intake and body weight tal cortices. Te comparison between KS and AUD regarding were measured all along the experiment and did not show the GM volume in the FCC and parietal cortices revealed any diference between groups (~6 ml/mouse/day). Results: that they were atrophied to the same extent, suggesting that behavioral alterations were signifcant after 6 months of BEARNI is sensible to the severity of alcohol-related GM risky alcohol consumption, as revealed by persistent memory abnormalities. Within the PC, the volume of the parahippo- impairments. After 9 and 12 months of alcohol exposure, campal gyrus correlated with the fuency score and was the balance abilities were gradually altered in the two groups only region to be specifcally atrophied in KS, suggesting that with risky alcohol consumption. Anxiety levels did not difer BEARNI is sensible to specifc brain abnormalities occur- between groups at any time. Brain volumes in various regions ring in KS. It is worthwhile noting that BEARNI remains a classically afected by alcohol consumption did not show any screening test and should not be considered as a sufcient tool between-group diference after 6, 9 or 12 months of alcohol to diagnose KS. An extensive neuropsychological assessment exposure. Conclusion: our results show that risky alcohol associated with a follow-up examination (in order to confrm consumption, even when not reaching AUD, drives a series the persistence of the neuropsychological impairments) is of behavioral alterations which are less severe but compatible required for a confrmed KS diagnosis. with the defcits described in AUD patients. Interestingly, Is impulsivity related to executive deficits in these defcits do not seem to be related to macrostructural patients with Alcohol Use Disorder? brain alterations. Microscopic analyses to study neuronal Nicolas Cabé1, Alice Laniepce1, Céline Boudehent1, Francis density, microgliosis and astrogliosis that could explain the 1 1 1 behavioral defcits observed are ongoing. Eustache , François Vabret , Hélène Beaunieux , Anne-Lise Pitel1 (1 Caen, France) BEARNI as screening tool for neuropsychological impairments and brain shrinkage in alcohol use Introduction: impulsivity, strongly associated with Alcohol disorder and Korsakoff’s patients Use Disorder (AUD), is a multidimensional construct encom- 1 1 1 Ludivine Ritz , Shailendra Segobin , Coralie Lannuzel , passing various diferent cognitive and behavioral components. Céline Boudehent1, François Vabret1, Anne-Lise Pitel1, Hélène Impulsivity is involved in the induction of the frst alcohol Beaunieux1 (1 Caen, France) consumption, reactivity to alcohol stimuli, loss of control over alcohol consumption, development of dependence, risk of Chronic and excessive alcohol consumption results in Alcohol relapse, and craving (1, 2). Anxiety and depression are also Use Disorder (AUD) without neurological complications and associated with craving and risk of relapse and may be related Korsakof ’s syndrome (KS) when combined with thiamine to impulsivity (3, 4). In clinical practice, impulsivity is assessed deficiency. These two clinical forms are accompanied by by self-questionnaires such as the Barratt Impulsiveness Scale widespread structural brain damage in both the fronto-cere- (BIS), which has been designed to measure trait impulsiveness bellar and Papez circuits as well as in the parietal cortex. Grey and their dimensions. Some authors rather consider impulsi- matter (GM) shrinkage in these brain regions is in agreement vity as a result of an executive dysfunction and more particu- with the neuropsychological impairments observed in AUD larly as an inhibition failure. Chronic and excessive alcohol patients early in abstinence including notably executive and consumption is indeed known to be associated with cognitive motor defcits as well as episodic memory disorders. AUD impairments and especially with executive defcits such as alte- and KS can be distinguished on the severity of the brain red planning, fexibility or inhibition (5). Impulsivity observed damage and cognitive defcits. Te main specifcity of KS is a in AUD patients may thus refect the behavioral consequences disproportionately encoding defcit in episodic memory, whose of impaired executive abilities related to a history of excessive severity allows distinguishing KS from AUD. BEARNI is a and chronic alcohol consumption. A better understanding of screening tool especially designed to detect neuropsychologi- the cognitive substrates of impulsivity is crucial to ofer appro- cal impairments in AUD. But the relevance of BEARNI for priate care when impulsivity is considered as a therapeutic

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 279 Congress target. Our aim was to investigate the relationships between fndings suggest that impulsivity would correspond to the impulsivity and executive functions in AUD patients, taking hyperactivation of the afective system rather than to the alte- patients’ alcohol history, as well as anxiety and depression, into ration of the refective system. Another explanation could be account. Considering that impulsivity is a consequence of the that AUD patients have difculty to self-evaluate their level dysexecutive syndrome, we would expect the impulsivity score of impulsivity when asked through questionnaires (7). Further to be related to executive abilities and alcohol exposure. If we studies are required to explore whether our fndings could be rather consider impulsivity as a matter of personality or emo- replicated using other impulsivity questionnaires (like UPPS tions, we would expect it to be related to thymic variables such or BIS 11) and executive tasks targeting other functions such as anxiety and depression, as well as early alcohol consump- as decision-making or planning. Clinically, these results sug- tion. Method: eighty-fve recently detoxifed AUD patients gest that impulsivity and executive abilities should be evalua- and sixty-three healthy controls (HC) matched for age, edu- ted and managed separately and in complementary perspective cation, and sex were included. Sociodemographic data and to prevent or reduce the relapse risk in AUD patients. information about patients’ alcohol consumption history were References collected. Anxiety was measured by the State-Trait Anxiety 1 - Lejuez CW, Magidson JF, Mitchell SH, Sinha R, Stevens MC, de Wit Inventory (STAI), and depression by the Beck Depression H. Behavioral and biological indicators of impulsivity in the develop- ment of alcohol use, problems, and disorders. Alcohol Clin Exp Res. août Inventory (BDI). Executive functions were assessed with 2010;34(8):1334-45. an extensive neuropsychological battery, which explored 2 - Perry JL, Carroll ME. The role of impulsive behavior in drug abuse. fexibility, inhibition, manipulation of information stored in Psychopharmacology (Berl). sept 2008;200(1):1-26. working memory, organization and strategy. Impulsivity was 3 - Sinha R, Fox HC, Hong KA, Bergquist K, Bhagwagar Z, Siedlarz KM. measured by the Barratt Impulsiveness Scale 10 (BIS10) Enhanced negative emotion and alcohol craving, and altered physiological responses following stress and cue exposure in alcohol dependent indivi- which is the latest validated version of this questionnaire in duals. Neuropsychopharmacology. 2009;34(5):1198. French. Parametric statistical analyses (Student’s T-tests) and 4 - Stanford MS, Mathias CW, Dougherty DM, Lake SL, Anderson NE, chi-square tests when appropriate were used to compare AUD Patton JH. Fifty years of the Barratt Impulsiveness Scale: an update and and HC groups. We then conducted correlational analyses review. Personality and Individual Differences. oct 2009;47(5):385-95. 5 - Oscar-Berman M, Valmas MM, Sawyer KS, Ruiz SM, Luhar RB, Gravitz (Bravais-Pearson) to investigate the relationships between ZR. Profiles of impaired, spared, and recovered neuropsychologic processes BIS 10 total score, cognitive abilities including executive in alcoholism. Handb Clin Neurol. 2014;125:183-210. functions, anxiety and depression, and alcohol history or 6 - Wiers RW, Gladwin TE, Hofmann W, Salemink E, Ridderinkhof KR. Cognitive bias modification and cognitive control training in addiction clinical variables. Results: compared to HC, AUD patients and related psychopathology: mechanisms, clinical perspectives, and ways were signifcantly more anxious and depressed and presented a forward. Clinical Psychological Science. avr 2013;1(2):192-212. more frequent alcohol family history. Moreover, AUD patients 7 - Le Berre A-P, Sullivan EV. Anosognosia for memory impairment in addic- were signifcantly more impulsive than HC as indicated by tion: insights from neuroimaging and neuropsychological assessment of their BIS 10 total score. AUD patients were impaired on metamemory. Neuropsychol Rev. 2016;26(4):420-431. all the executive tests used in the study except on the verbal A smartphone App to assess alcohol consumption fuency task. Teir speed processing was signifcantly lower behaviours: development, validity, compliance, than in HC. Correlations revealed that in AUD patients, the and reactivity BIS 10 total score correlated with none of the executive and 1 1 1 alcohol history measures. By contrast, impulsivity correlated Antoinette Poulton , Jason Pan , Loren Richard Bruns , Richard 1 1 1 with higher anxiety and depression. Discussion: AUD patients O Sinnott , Robert Hester ( Parkville, Australia) presented a high level of impulsivity, which was related nei- ther to their executive defcits nor to their history of alcohol Background: Although research into problem drinking often consumption. Our fndings indirectly indicate that impulsivity, relies on retrospective measures to assess alcohol intake, such as evaluated by the BIS 10, does not seem to be a consequence methods have been found to distort actual consumption of an alcohol-related dysexecutive syndrome. Impulsivity levels/patterns. Real-time electronic protocols in the form of could rather be a vulnerability factor, linked to affective smartphone apps are consequently advocated. Tere is limited parameters (anxiety, depression) and potentially favoring the research pertaining to the development, validity, compliance, development of excessive and chronic drinking, which in turn and reactivity of using such apps in the experimental arena. would result in altered executive functions. Tese unexpected Methods: an iterative process guided the development of results could be interpreted within the theoretical framework the CNLab-A app. Healthy individuals (N=671) completed of the neurocognitive dual-process model, which proposes demographic questions and a 21-day Timeline Followback that addictions may be the product of an imbalance between before using CNLab-A for 21 days. We considered the size 1) a “refective system”, involved in the cognitive evaluation and diversity of the sample; compared data reported via of the stimuli by means of memory and executive functions, retrospective measures with that captured using CNLab-A; responsible for controlled-deliberate responses and 2) an and, assessed the data for evidence of compliance and reacti- “affective-automatic system”, involved in the emotional vity as a function of hazard versus non-hazard drinker status. evaluation of the stimuli, initiating automatic-appetitive Results: CNLab-A yielded a large and diverse sample. On responses. Te imbalance between these two systems would average, participants submitted data on 20.27 out of 21 days. account for rapid decision making, prioritizing short-term Compared to Timeline Followback, a signifcantly greater reward irrespective of the long-term consequences (6). Our percentage of drinking days (24.79% vs. 26.44%) and signi-

Alcoologie et Addictologie. 2019 ; 41 (3) : 232-282 280 Congress fcantly higher total intake (20.30 vs. 24.26 standard drinks) showed less difculties in emotion regulation (DERS-F) for was recorded via the app. Both hazard and non-hazard drin- all individuals immediately after (T1 ART) or six months kers were highly compliant with app protocols. Linear growth after the end of therapy (T2 ART) than before ART session analyses revealed hazardous drinkers decreased their alcohol (T0 ART). Compared to control group, individuals with intake by 0.80 standard drinks over the 21-day experimental emotion training partially recovered on DERS-F subscales period. Tere was no change to the drinking of non-hazard especially for strategies and emotional clarity. In addition, individuals. Both hazard and non-hazard drinkers showed individuals had higher levels of non-adaptive strategies for a slight decrease in responding (“yes”) to drinking behavior emotion regulation (CERQ) before (T0 ART) than after over the same period. Conclusions: smartphone apps appear therapy session (T1 ART, T2 ART). In comparison to control an efective and methodologically sound means of obtaining group (T0 control), they had less non-adaptive strategies after alcohol consumption information. ART session (T1 ART). Finally, mindfulness total score was higher after therapy session in ART group (T1 ART) than Affect regulation training (ART) program in in control group (T0 control, T1 control). Conclusion: these alcohol abstinence consolidation results demonstrated a recovery of emotion regulation abili- Caroline Claisse1, Marie-Charlotte Gandolphe1, Lydie Defrance1, 2 1 1 2 ties after emotional training in a sample of abstinent alcohol Mickael Naassila , Jean-Louis Nandrino ( Lille, France, Amiens, individuals. Compared to Control group, the ART group France) presented less difculties in emotional abilities and regulation strategies, and higher levels of mindfulness. Te ART program Background: this study aimed at exploring the evolution allows greater fexibility in the emotion regulation strategies of emotion regulation strategies and abilities with training involved in consolidating abstinence and should be used both remediation program in abstinent patients sufering from initially at the beginning of withdrawal but also in individuals severe alcohol use disorders (AUD). Te main objective of this in the process of maintaining their abstinence. project is the consolidation of abstinence with the acquisition or reinforcement of new emotion regulation strategies. In Effect of inflammatory pain on alcohol induced the Adaptive Coping with Emotions Model (ACE Model, dopamine release in the NAc and alcohol relapse Berking & Whitley, 2014), adaptive emotion regulation is in rats 1 1 conceptualized as a situation-dependent interaction between Yolanda Campos-Jurado , Jose Luís González-Romero , Jesús emotion regulation skills. To this end, the Afect Regulation Lorente1, Ana Polache1, Luis Granero1, Teodoro Zornoza1, Lucía Training (ART, Berking & Whitley, 2014) was conceptua- Hipólito1 (1 València, Spain) lized in sixe training modules (psycho-education, muscle and breathing relaxation, non judgmental awareness, acceptance Epidemiologic data have shown a relationship between pain and tolerance, compassionate self-support, analyzing emotions and addiction especially to and alcohol. Indeed, a and modifying emotions) related to these emotion regulation recent clinical study undercovered that the correct manage- skills. Each unit focuses on one emotion regulation skill. ment of pain in patients with a previous history of alcohol Method: 117 alcohol-abstinent individuals abstinent from use disorder decreases the risk of relapse in alcohol drinking, two weeks to several years were recruited in day hospitals and suggesting that in this prone population, pain may increase groups of Alcoholics Anonymous association. 87 individuals the vulnerability to relapse in alcohol consumption. Previous participated in the ART program and were compared to 30 data in rats revealed that infammatory pain desensitizes alcohol-abstinent individuals who do not have the program. mu opioid receptors (MORs) in the ventral tegmental area Te ART program consisted of six weekly 3-hour therapy (VTA) and increases intake of high doses of heroine. Due to sessions in a group of 5 to 8 persons. In control group (n=30), the relevant role of MORs in alcohol efects, we hypothesize evaluations were proposed within therapy session. For all indi- that this desensitization may also alter the pattern of acti- viduals, the evaluation consisted of two parts: a clinical ques- vation of the mesocorticolimbic system exerted by alcohol tionnaire (concerning their current situation, alcohol use and and therefore have an efect on alcohol relapse. In our study, context) and a cognitive and emotional assessment. Drinking we evaluated the efect of infammatory pain on accumbal history and emotion regulation processes were assessed using increase of dopamine release elicited by 1.5 g/kg of ethanol the Difculties in Emotion Regulation Scale (DERS-F) and (s.c.). Tis microdialysis study showed that the presence of the Cognitive Emotion Regulation Questionnaire (CERQ). infammatory pain blunted the increase of extracellular dopa- Anxiety and depression scores were assessed using Hospital mine levels in the Nucleus Accumbens induced by ethanol. Anxiety Depression Scale (HADS). Finally, mindfulness levels Later on, we evaluated the efect of infammatory pain on were assessed with Five Facets Mindfulness Questionnaire the alcohol deprivation efect (ADE) in long-term ethanol- (FFMQ). For the experimental group, the same evaluation experienced rats. After four cycles of free ethanol intake was performed before ART program (T0 ART), after the and abstinence periods, infammatory pain did not afect to ART program (T1 ART) and six months later (T2 ART). the magnitude of the ADE. Tese data further support the In control group, the evaluation was performed twice, one at impact of pain on the neurochemical events on the dopami- (T0 control) and six weeks later (T1 control). Result: dife- nergic mesocorticolimbic system following alcohol adminis- rences in emotion regulation strategies were found according tration and also underscore the necessity of fnding an appro- to the participation in ART remediation program. Te results priate paradigm to determine the behavioral consequences.

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Emotional memory in young binge drinkers signifcant diferences between patients in withdrawal and 1 2 2 Carina Carbia , Montserrat Corral , Francisco Caamaño-Isorna , after 2 weeks, among them in genes which have been repor- 2 1 2 Fernando Cadaveira ( Cork, Ireland, Santiago de Compostela, ted to play a role in withdrawal symptomatology in previous Spain) studies (SLC29A1, FYN). As expected, methylation between patients and controls difered considerably, also in genes Background: college binge drinking (BD) has been linked to implicated in withdrawal (FKBP5, BDNF, EFNA5). Search persistent cognitive difculties, especially in episodic memory. for diferentially methylated regions and gene ontology based However, despite impairments in emotional functioning have gene set analysis revealed involvement of apoptotic processes been associated with the development of alcohol use disorders, in acute withdrawal. Tis has been also shown in other assess- the emotional sphere has been relatively unexplored in BDs. ments with alcoholic patients. Tis epigenome-wide longitu- Te purpose of this study is to examine the efects of BD in dinal methylation study conducted in the so far largest sample emotional episodic memory. Methods: a cohort of 180 (96 ) of severely alcohol-dependent individuals sufering from healthy college students was followed during two years (18-20 withdrawal symptoms replicates known and suggests novel years old) and their alcohol use was recorded. In the last assess- genes, which may play a crucial role in alcohol withdrawal. ment, participants completed an adaptation of the Emotional Alcohol consumption during pregnancy: Verbal Learning Test (EVLT). Generalized linear mixed preliminary data on the effects of environment models were applied. Te models were adjusted by psycho- pathological symptoms (BSI-18). The neuropsychological enrichment on transcriptional regulation of analyses were carried out separately for males and females, relevant key genes in mothers and offspring 1 2 1 2 in accordance with sex diferences in the development of F. Bellia , A. Wille-Bille , M. Pucci , R.S. Miranda-Morales , emotion circuitry in adolescents. Results: in females, BD was R.M. Pautassi2, C. D’Addario C1,3 (1 Teramo, Italy, 2 Córdoba, associated with poor performance in the emotional memory Argentina, 3 Stockholm, Sweden) task, in particular lower recall of neutral words and greater recall of negative versus neutral words. Whereas in males, no Introduction: the consumption of alcohol by mothers during alcohol-related efects were found. Conclusions: females binge pregnancy may lead to mental or physical issues for the new- drinkers present difculties in emotional episodic memory lin- born, as well as be potentially dangerous to themselves after ked to the interference of negative content. Tis is in line both delivery. possibly increasing the risk of alcohol use due to with sex-related diferences in the recall of emotional memory heightened stress. Te diferent phenotypes occurring in both and an alcohol-related aberrant processing for emotionally mothers and ofspring might involve the epigenetic regulation salient stimuli, which might result in greater vulnerability of genes transcription. Using an animal model of prenatal etha- to afective disturbances among women. Further research is nol exposure, we here studied in mothers postpartum and in needed to understand the role of emotional functioning in the their ofspring the efects of brain transcriptional regulation of escalation of alcohol abuse, from a gender perspective. target genes and how environmental enrichment might modu- late possible alterations. Methods: the dams were given one Methylation profiles during acute alcohol daily intragastrically administration of 0.015 ml/g of a 16.8% withdrawal in a clinical sample 1 1 1 1 v/v ethanol solution or a similar volume of vehicle (gestational Lea Sirignano , Stephanie H Witt , Josef Frank , Jens Treutlein , days 17-20). After delivery litters were divided in two groups 1 1 1 Fabian Streit , Ulrich Frischknecht , Jerome C Foo , Franziska to distinguish infancy from adolescence. Starting from PD14 2 3 4 3 Degenhardt , Gabi Koller , Ulrich Preuss , Peter Zill , Kristina in both infants and adolescents were evaluated anxiety-like 3 2 1 1 Adorjan , Markus Nöthen , Rainer Spanagel , Falk Kiefer , behavior and exploratory activity together with risk-taking Marcella Rietschel1 (1 Mannheim, Germany, 2 Bonn, Germany, behavior in the light-dark box and in the concentric square 3 Munich, Germany, 4 Herborn and Halle-Wittenberg, Germany) feld test respectively. Mothers were sacrifced 21 days after delivery by decapitation, ofspring were sacrifced one day after Withdrawal is a serious and sometimes life threatening event the behavioral tests (PD32), brain dissected and nucleic acids in alcohol-dependent individuals. It has been suggested, extracted for genes expression studies and genes promoter eva- that epigenetic processes may play a role in this context. luation of DNA methylation/hydroxymethylation. Results and Identifcation of genes involved in such processes may hint to discussion: our fndings so far show selective altered expression relevant mechanisms underlying withdrawal. In the present for BDNF and prodynorhin genes in the VTA of adolescent study we sought to longitudinally investigate epigenome- rats prenatally exposed to alcohol and of dams exposed to wide methylation patterns in 100 severely alcohol-dependent alcohol during pregnancy, whereas environmental enrichment patients during alcohol withdrawal and after 2 weeks of reco- partially reverted these changes at least in adolescent ofspring. very, and also in 100 matched controls. More than 850,000 Moreover, altered methylation at specifc CpG sites at both methylation sites were assessed using Illumina EPIC bead gene promoters was observed consistently with the changes chips. Refecting the high quality of our methylation data, in genes transcription. Tese data, even if preliminary, might we found – consistent with earlier reports – that correlation be promising in order to understand the protective role of of methylation age with biological age of assessed individuals environmental switch on the efects evoked by alcohol also was very high (r=0.9). We found pronounced genome-wide suggesting molecular mechanisms accounting for it.

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Pr Mickael Naassila* * President 17th ESBRA Meeting

17th European Society for Biomedical Research on Alcoholism congress 21-24 September 2019, Lille – Posters abstracts

Self-estimation of blood alcohol concentration in correction of under-estimated BAC could improve awareness patients admitted in emergency department in AUD patients and elicit motivational changes. Stéphanie Alaux-Cantin1, Bernard Angerville1, Mickael Naassila1, Alain Dervaux1 (1 Amiens, France) The role of the gut-brain axis in alcohol dependence: design of the Gut2Brain study Studies conducted in general population reported that alco- Camille Amadieu1, Sophie Leclercq1, Peter Stärkel1,2, hol drinkers under-estimate their blood alcohol concentration Philippe de Timary1, Nathalie M. Delzenne1 (1 Louvain, Belgium, (BAC). However, no study has investigated the accuracy of 2 Brussels, Belgium) BAC self-estimation in alcohol intoxicated patients admitted in Emergency Department (ED). Rationale: Te gut microbiota, a huge community of micro- To assess this question, all consecutive patients admitted organisms (comprising bacteria, viruses, fungi and yeast) in ED of the University Hospital of Amiens, France, with living in our intestine, has been shown to regulate many at least BAC of 0.6 g/L were included in the study. Self- important functions for human health including metabolism, estimated BAC was assessed using a visual analogic scale, immunity as well as brain functions and behavior. Our pre- compared with objective measurements of BAC. We next vious studies have shown that chronic alcohol abuse induced performed comparisons between moderate, mild or severe a leaky gut and alterations in the composition of the gut mi- AUD patients according to the DSM-5 criteria. We assessed crobiota, which are correlated with psychological symptoms the subjective efects of alcohol (SEA) using SEA scale. such as depression, anxiety and alcohol craving, suggesting Preliminary results showed that patients included in the pre- the involvement of the gut-brain axis in the development of sent study under-estimated their BAC (-0.6±0.3 g/L, n=20). alcohol use disorders (AUD). Interestingly, moderate AUD patients over-estimated BAC Te Gut2Brain study aims at modulating the gut-brain axis (0.5±0.5 g/L, n=3), mild AUD patients correctly estimated of AUD patients by administering dietary fbers with prebio- BAC (0.0±0.3 g/L, n=5), and severe AUD patients clearly tic properties which are known to modify the composition of under-estimated BAC (-1.1±0.3 g/L, n=12). Te sensitivity to the gut microbiota. the positive sub-scale of the SEA (i.e. lively, funny, talkative) Methodology of the Gut2brain study: Tis is a randomized, was important in moderate AUD, intermediate in mild AUD double-blind, placebo controlled study including 50 patients. and low in severe AUD patients except regarding low arousal Twenty-fve patients are assigned to the prebiotics group positive efects that remain relatively important (i.e. relaxed, and 25 patients are in the placebo group. AUD patients are calm, mellow). Both groups displayed a low sensitivity toward hospitalized for a 3-week detoxifcation program in the alco- negative alcohol efects (i.e. aggressive, rude, woozy). hol withdrawal unit of St Luc academic hospital (Brussels, Taken together, our preliminary results showed that severe Belgium). AUD patients under-estimated BAC and displayed an altered Biological (microbiota composition, bacterial metabolites, sensitivity to the SEA. Further investigations are necessary inflammatory markers) and psychological measurements to assess whether such a brief intervention including the (depression, anxiety, craving, sociability) have been performed

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 368 Congrès twice, at the onset of alcohol withdrawal (T1 = before starting with a normal day without alcohol consumption, suggesting the prebiotic treatment) and at the end of the detoxifcation that a single BD episode may signifcantly compromise the program (T2 = after 17 days of prebiotics supplementation). allocation of attentional resources needed to perform the task Because microbiota composition is heavily influenced by in the following day. Additionally, after a night engaging in nutrition, diet anamnesis have been handled to evaluate the BD, students displayed a marginally signifcant decreased P3 nutritional habits of AUD patients with a special focus on and LPC amplitude. Although still tentatively, these results fber intakes. could indicate that a BD session may lead to impairments Conclusion: Te Gut2Brain study will investigate for the frst on attentional and working memory processes in young BDs. time the efects of prebiotics on gut microbiota composition and function, systemic infammation and psychological symp- Gene expression changes associated with toms of AUD patients. Te results of this study will help to stress-induced alcohol escalation design new therapeutic and/or preventive targets for AUD Estelle Barbier1, Riccardo Barchiesi1, Kanat Chanthongdee1, patients. Markus Heilig1 (1 Linköping, Sweden)

The effects of hangover on inhibitory control in Comorbidity of alcohol use and anxiety disorders is a major young binge drinkers: An event-related potentials cause of disability and a challenge for mental health services. study Both disorders are characterized by broad and persistent Natália Antunes1, Rui Rodrigues1, Alberto Crego1, changes in gene expression within brain areas involved in Eduardo López-Caneda1 (1 Braga, Portugal) regulation of negative afect including the prefrontal cortex and the amygdala. However, the shared underlying mecha- During adolescence and at the time of entering university, nisms are still not well known. young people seek new sensations and they are more likely to In our study, we used a rat model of social defeat stress (SDS) engage in high-risk behaviours, such as drug abuse. Alcohol to assess the impact on alcohol- and anxiety-like behaviors. besides being the most consumed drug in the world, it also In addition, a second group of rats were made to witness represents the third higher risk factor for disease and largely the SDS in order to unravel the psychological component contributes to the number of deaths worldwide. Te excessive from the combined physical and psychological stress in alcohol use can lead to a pattern known as binge drinking the defeated animals. In accordance with previous studies, (BD), which is characterised by heavy alcohol intake over we found individual variability in the behavioral outcomes a short time, followed by periods of abstinence. Tis form following social stress. Stress induced by social defeat or by of alcohol misuse has received special attention in the last witnessing SDS, led to an increase in operant alcohol self- decade mainly due to its high prevalence among youngsters administration and anxiety-like behaviors only in a subset and the negative consequences resulting from that. of animals. Behavioral studies were performed ten days One of the major consequences immediately after a BD after the last social defeat, suggesting a long lasting efect of episode is the hangover experience. Hangover, strictly related the social stressors on both alcohol intake and anxiety-like to BD, can be described as a series of unpleasant physical behaviors. Gene expression changes observed on the subset and mental symptoms, which follow the intake of large of rats showing both alcohol and anxiety-related behaviors amounts of alcohol and are especially signifcant when the are assessed using our custom made NanoString panel. Our blood alcohol concentration reaches 0 g/dL. Some studies panel comprises about 400 genes involved in critical neuro- have demonstrated that alcohol hangover may afect cogni- nal functions such as neurotransmitter release and synaptic tive functioning, namely memory, attention and psychomotor plasticity. It also include epigenetic regulators. Tis is of par- performance. Nevertheless, to the best of our knowledge, ticular interest as stress and heavy alcohol have been shown no study has been conducted with the aim of assessing the to reprogram the transcriptome, making interventions that behavioural and electrophysiological consequences of alcohol target epigenetic mechanisms an attractive novel approach to hangover after a typical BD episode despite the important develop therapeutics. implications that might result from this research. Aiming Investigating the role of neuroinflammation in to understand how inhibitory control may be afected the day after a single BD session, behavioural measurements Long-Term Depression impairment induced by two and brain activity recorded from 64 electrodes were analysed ethanol binge exposures (TEBE) in young rats while 10 college BDs (six females) performed a Go/NoGo Cédric Bertrand1, Catherine Vilpoux1, Mickael Naassila1, task before and after a typical BD night. Te reaction times; Olivier Pierrefiche1, Stéphane Peineau1 (1 Amiens, France) percentages of correct responses and correct inhibitions; the amplitude and latency of P2, N2, P3 and Late Positive Binge Drinking (BD), an alcohol consumption pattern des- Component (LPC) were assessed. cribed as a fast way to reach drunkenness, is associated to Despite having found no hangover efects at the behavioural many cognitive impairments including hippocampus related level, electrophysiological abnormalities emerged the day after memory issues and leads to chronic neuroinfammation with a heavy alcohol drinking episode. Specifcally, decreased P2 deleterious efect at the neuronal level. Hippocampus neuro- amplitudes were observed after a BD night in comparison nal plasticity mechanisms, such as Long Term Depressions

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(LTD), are crucial in learning and memory processes and point of the progressive ratio were analyzed. modulated by neuroinfammation factors. Our results revealed that animals in a ketosis state exhibited, We used young rat to model the early steps of BD consump- in general, a trend towards a decrease in ethanol consumption tion on cognitive deficits (hippocampus slices, plasticity in terms of the FR1 and FR3, but did not show changes in recordings 48h after TEBE 3g ethanol/kg bodyweight, i.p. their motivation to drink compared to animals fed a standard given 9h apart; Silvestre de Ferron et al., 2015) and found diet. an abolition of population spike LTD in CA1 neurons, We propose that future investigations are necessary to clarify concomitant with learning impairment. We now investigated which neuroadaptations underlie the efects produced by the the impact of TEBE on synaptic LTD with patch-clamp KD. Our results suggest that the nutritional state is a useful techniques and investigated the role of neuroinfammation. tool for the future treatment of alcohol use disorders. We found a partial synaptic LTD inhibition, suggesting that Acknowledgements: Generalitat Valenciana, Conselleria population-spike LTD abolition could be due to an efect Educación, Dirección General de Universidades. Grupos of TEBE at the synaptic level. We then hypothesized that de Investigación de Excelencia. Prometeo 2018/132. synaptic LTD inhibition originates from an emerging neuro- Instituto de Salud Carlos III, Red de Trastornos Adictivos inflammation. We analyzed microglial cells morphology (RD16/0017/0007) y Unión Europea, Fondos FEDER “una and infammatory markers with MACS cell separation and manera de hacer Europa”. RT-qPCR technique after TEBE. In parallel we treated Time course and specificity of attentional bias in TEBE rats with anti-infammatory drugs. When applied before the frst exposure, anti-infammatory treatment en- binge drinking: An eye-tracking 1 1 hances synaptic LTD impairment but not if rats are treated Zoé Bollen ( Louvain, Belgium) just before the second exposure. Tese results highlight a neuroprotective role of neuroinfammation in the early step Attentional bias is a core characteristic of alcohol use of binge drinking episodes, in contrary to its deleterious role disorders (AUD), playing a crucial role in their development in multiple episodes of binge drinking. and persistence. Many behavioural studies have showed an increased allocation of attention towards alcohol cues in pa- Ketosis modulates alcohol consumption in adult tients with AUD and revealed a direct link between bias and male mice craving, alcohol consumption or relapse risk. Nevertheless, M.Carmen Blanco-Gandía1,2, Francisco Ródenas1, its underlying mechanisms are still poorly understood. Eye- Marina D. Reguilón1, José Miñarro1, Marta Rodríguez-Arias1 tracking measures, ofering deeper insights regarding the timeline of the bias, constitutes an innovative way to renew (1 Valencia, Spain, 2 Teruel, Spain) its exploration. Te present study used eye-tracking measures to: (a) inves- In recent studies, metabolic or nutritional treatments for dif- tigate attentional bias in a subclinical AUD population (i.e. ferent disorders, such as epilepsy, Alzheimer’s disease, cancer binge drinking), (b) determine the time course of the bias, by or autism, have proved to be successful. Te ketogenic diet disentangling the early from late processing stages, (c) clarify (KD) is a high‐fat diet, low in carbohydrates and balanced the specifcity of the bias towards alcohol or its generalization in proteins, that induces changes in the body’s main energy towards other appetitive stimulations (i.e. food stimuli), and source, since it uses ketone bodies instead of glucose. Te (d) explore the relation between craving and attentional bias. KD has been linked to the amelioration of all the above‐ Two groups of participants (42 binge drinkers, 43 controls) mentioned conditions, but the mechanisms underlying its performed a visual probe task, which requires the detection of therapeutic effects are still unclear. On the other hand, an arrow preceded by pictures from diferent conditions: (1) several recent studies have suggested that the type of diet alcohol vs. soft, (2) alcohol vs. food, (3) salty or sugary food (for example palatable food or cafeteria diet) and the way it vs. healthy food. Eye-tracking measures highlights the pre- is consumed (continuous access or binge eating) modulate sence of a bias towards soft and healthy food among control the development of drug addiction. For instance, high‐fat participants. Complementary analyses indicated that binge and ‐sugar diets increase cocaine and ethanol consumption in drinkers with high level of craving showed a bias towards mice, as well as their sensitivity to the conditioned rewarding alcohol and high-caloric food, unlike those with a low level efects of both drugs. Te present work aimed to study if the of craving. Te alcohol attentional bias is thus neither related KD can modulate the rewarding efects of alcohol and to to binge drinking, but rather to the association between this assess its potential as a therapeutic target to decrease alcohol drinking pattern and craving. consumption. A total of 30 adult male mice of the OF1 strain (PND 42) Fetal alcohol syndrome prevention in women: were assigned either a standard diet (n=14) or a Ketogenic Attitude to pregnancy Diet (KD) (n=16). When a ketosis state had been sustained Ulia O. Remenyuk1, Ekaterina A. Burina1 (1 Saint Petersburg, for 7 days, the reinforcing and motivating efects of ethanol Russia) were measured by means of the oral self‐ administration pa- radigm, in which the number of reinforced responses (Fixed A number of studies addresses fetal alcohol syndrome (FAS) Ratio 1 and 3), ethanol consumption (g/kg) and the breaking prevention, but almost none investigate FAS aspects in

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 370 Congrès connection to pregnancy attitude. Awareness helps a woman tially methylated regions (DMRs) between controls and to develop an adequate relation to pregnancy that leads to a AUD subjects. We identifed a total of 914 DMRs (26% healthy behavior (Health Belief Model, Rosenstock I., 1974). hypomethylated and 74% hypermethylated), with CpGs Terefore, a pilot study was conducted. ranging in diferential methylation from ~5-43%. As we Sample: 35 non-pregnant women of childbearing age (M=27), observed in our prior nonhuman primate NAcc GWDM never been pregnant and able to give birth. analysis, the majority of DMRs mapped intragenic locations Methods: screening; informed consent; interview (pregnancy (p(hypergeometric) < 1.02e-288), mostly to intronic regions, while attitude, FAS awareness, alcohol-exposed pregnancy (AEP) only 5% of the DMRs mapped to promoters. In terms of risk); personal inventory “Big 5”; personal time perspective proximity to CpG islands, most of the DMRs overlapped measure; subjective control evaluation method. Primary with CpG islands (44%; p(hypergeometric) = 7.10e-1152), followed preventive measures were realized with every participant. by open sea (30%; p(hypergeometric) = 7.40e-502) and CpG island Analysis: statistical methods, “R Studio”. shores (18%; p(hypergeometric) = 2.10e-122). Cell-type enrich- Study results revealed very poor FAS awareness – just 34% of ment analysis of the 707 DMRs mapping to genes or their women heard about the syndrome but only 14% of them gave promoters showed that chronic alcohol consumption signif- correct answers. AEP-risk was observed in 26% with average cantly (hypergeometric test) afects the methylome of genes 4 alcohol drinks a time, and contraception risk – in 31%. specifc of astrocytes (Astroc, n=56; p=8.9e-10), endothelial Personal features of participants with AEP-risk showed cells (Endot, n=61; p=4.7e-12), microglia (Microg, n=54; average results with a tendency of women with low/normal p=6.3e-92), oligodendrocytes (Oligod, n=57; p=3.2e-10) and alcohol use to have higher readiness for cooperation (U=62.5; neurons (Neur, n=66; p=1.6e-14). Using Ingenuity pathway p=0.0406). Subjective control was found on average level and analysis, the following pathways were enriched: axonal was not related to at-risk behavior. Time perspective results guidance signaling (19 genes, p= 5.5e-3), Wnt/B-catenin indicated high expectations about participants’ future. signaling (19 genes, p=1.4e-6), synaptic long term depression (18 genes, p=1.9e-5), CREB signaling in neurons (18 genes, Attitude to pregnancy was divided into categories: positive p=1.5e-4), gap junction signaling (17 genes, p=2.6e-4), cor- (45.5%), negative (30%), neutral (24.5%). No relation to ticotropin releasing hormone signaling (13 genes, p=4.2e-4), other characteristics studied was observed, possibly due to a dopamine cAMP signaling (14 genes, p=4.6e-4), opioid small sample. Identifed categories can help in determining signaling pathway (18 genes, p=6.6e-4) and synaptogene- woman’s pregnancy attitude in order to adjust behavior to sis signaling pathway (19 genes, p=5.5e-3). Some of these more health-saving. genes have been previously associated with alcohol abuse (i.e Pilot study results can be useful for preventive programs AGAP1, SEMA5A), which reinforces the role of these genes design and further implementation. in modulating alcohol abuse as well as the potential of our Genome-wide DNA methylation analysis of the approach to not only identify these genes but also provide human postmortem nucleus accumbens identifies important details on the underlying epigenetic mechanisms differential methylation in AUD individuals potentially regulating their activity in the context of alcohol 1 1 abuse. Other genes identifed in this study have not yet been Rita Cervera-Juanes ( Oregon, USA) linked to addiction but their function is highly relevant in modulating alcohol-associated neuronal adaptations (i.e. Chronic alcohol use has been linked to alterations in synap- TCF7L2 involved in Wnt signaling). Altogether, our data tic plasticity thought to contribute to alcohol dependence, suggests that a history of alcohol abuse is associated with tolerance, craving and withdrawal. Clarifying the genes diferential DNA methylation in afecting synaptic plasticity and regulatory mechanisms underlying such neuroadapta- mechanisms, similarly to what we observed in NHPs with tions is critical to fully understand and treat alcohol abuse. long-term consumption of heavy doses of alcohol. Tis study Here, we explore the methylome of the postmortem human not only provides genes, but equally important, it provides nucleus accumbens (NAcc) from controls and AUD subjects. epigenetic information on how these genes may be regulated To identify diferential methylation signals we conducted by alcohol, and how they could be targeted to revert such genome-wide DM (GWDM) analysis using the Agilent efect from a therapeutic perspective. SureSelect Human MethylSeq kit on NAcc tissue from age-matched pairs of AUD and control subjects (32 males) H2O2, a major reactive oxygen species of alcohol obtained from the New South Wales Brain Tissue Resource metabolism induces autophagy without involving Center (NSWBTRC). We obtained ~108 million raw reads mTOR per library that were aligned to the GRCh38/hg38 assembly Cheng Chen1, Teresa Peccerella1, Sebastian Mueller1, of the human genome using Bismark. After quality control 1 1 evaluation, we obtained ~2.5 million CpG sites per sample Vanessa Rausch ( Heidelberg, Germany) that were used for downstream analyses. Diferential methy- lation was analyzed using the generalized linear modeling Background and aims: Alcohol-mediated reactive oxygen approach implemented in RnBeads including covariates of species (ROS) formation in the liver, mainly H2O2, contri- interest (i.e. batch sequencing efect, age, smoking status, butes to disease progression and eventually hepatocellular etc.) and Comb-p analysis was used to identify diferen- carcinoma development in patients with ALD. Enhancement

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 371 Congrès or activation of autophagy, with the suppression of mTOR Health Questionnaire (GHQ), and the trained psychiatrists signaling, is likely to play an important role in early stages of examined the risk individuals with a Chinese version of the the alcoholic liver disease (ALD). However, with the progres- Structured Clinical Interview for Diagnostic and Statistical sion of the disease, the expression of mTOR increases dra- Manual IV axis I disorders. matically leading to the suppression of autophagy. It is also Results: Tere were 27,917 enrolled, and 27,489 completed in known, that H2O2 is involved in the regulation of autophagy this survey. Adjusted for gender, age and other demographic in both acute and chronic ALD models, however, the exact items, the one month prevalence of AUD was 5.27% (95%CI underlying molecular mechanisms are still unclear. Terefore, 5.01-5.54), and at the head of mental diseases’ prevalence, we investigated in vitro and in vivo by using alcohol mouse with the signifcant diference on gender (Z=45.29, p=0.00), model alterations in mTOR signaling as well as downstream but without the difference on residential place (Z=1.46, efects induced by H2O2 and low oxygen tension. p=0.14). Methods: Huh7 hepatoma cells and VL-17A cells (stably Conclusions: Te AUD prevalence in Shandong was high transfected with CYP2E1 and ADH) were cultured with the and should be highlighted as a public health problem. GOX/CAT system, which allows an independent control of hydrogen peroxide as well as oxygen levels, in combination Effect of trauma and family alcohol using with diferent doses of ethanol. LC3B, p62, mTOR and situations in childhood onthe occurrence of autophagy related proteins (e.g. AMPK, AKT, STAT3) were alcohol dependence in Chinese male patients analyzed by western blot. Same analyses were performed in Xu Chen1,2, Jiapei Yang1 (1 Binzhou, 2 Jinan, Shandong, People’s liver tissues of C57BL/6 mice treated with acute (alcohol Republic of China) binge) and chronic ethanol (20% Ethanol in the drinking water) for 4 weeks (n=4). Background and objective: It was proved by some studies Results: H2O2 signifcantly increased LC3B activation and that childhood traumas have strongly efect on the arousing this efect could be efciently blocked by N-acetyl cysteine of alcohol dependence in males, but the role of family alcohol (NAC), which is a ROS scavenger. Interestingly, even though using situations is unclear. Tis cross-section was aimed to the LC3B activation was increased by H2O2, the m-TOR analyze the efects of trauma and family alcohol using situa- expression was not suppressed as normally expected. Co- tions in childhood on the occurrence of alcohol dependence. treatment of hepatoma cells with H2O2 and the mTOR Methods: In this study, the questionnaires and the formu- inhibitor Rapamycin led to an increased autophagic fux as lated structure interview were used. 120 patients with alco- compared to single H2O2 and Rapamycin treatment. Te in hol dependence and 103 healthy volunteers were assessed vivo experiments showed a combined activation of LC3B by Childhood Trauma Questionnaire (CTQ), self-made and suppressed p62 and AKT levels as well as enhanced alcoholic using questionnaire for parents and the formulated p-AMPK expression in the livers of the acute alcohol group. interview. In contrast, mice exposed to chronic alcohol showed blocked Results: Te scores of CTQ, the fathers’ frequency of drin- autophagic fux with dramatically increased LC3-I and p62 king in the patients and inducing to drink in childhood were levels. signifcantly higher than those of volunteers; and the rate of Conclusion: Our fndings underscore an important role of parents’ opposed attitudes to drinking in patients were lower H2O2 in regulating autophagy during acute and chronic than that of the volunteers’. It was showed in the multiple- alcohol ALD exposure. Further studies will be needed to factors analysis that the relative risk factors of alcohol address the diferences between acute and chronic alcohol- dependence were Parents’ drinking frequency, No-opposed mediated efects as well as to identify H2O2-induced signa- attitude to drink, Being induced to drink and Lower-level ling pathways that regulate autophagy. education. Prevalence and demographic characteristics Conclusions: Compared to childhood trauma, parental alco- of alcohol use disorder in Chinese Shandong hol using is the more important role in the formation of provincial adultresidents: A cross-section alcohol dependence. epidemiologic survey Cross-modal processing of emotions in Severe Xu Chen1,2, Ruzhan Wang2, Yuchen Zhang1, Jingxuan Zhang2 alcohol use disorders: Impaired discrimination (1 Binzhou, 2 Jinan, Shandong, People’s Republic of China) of anger and fear under dynamic audiovisual Background and objectives: Tis survey aimed to investigate conditions 1 2 1 the prevalence and demographic characteristics of alcohol Coralie Creupelandt , Fabien D’Hondt , Federica Falargiarda , 1,3 1 1 use disorder (AUD) in Chinese Shandong provincial adult Olivier Collignon , Pierre Maurage ( Louvain-la-Neuve, residents. Belgium, 2 Lille, France, 3 Trento, Italy) Methods: Multistage stratifed random sampling was used to identify 34 urban communities and 62 rural adminis- Severe alcohol use disorders (SAUD) are associated with trative villages as the sampling sites in Shandong province, a large variety of affective disturbances, among which a with the 300 sample size of each site. Te trained psychia- well-established decoding defcit for facial and vocal emo- tric nurses completed the primary screening with General tional expressions. Tis defcit has recently been found to be

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 372 Congrès increased in cross-modal settings, namely when inputs from (NAc) in abstinence and alcohol relapse phase. No diferences diferent sensory modalities have to be combined. Compared were found in males. Nevertheless, we observed in females to unimodal emotional stimuli, cross-modal ones allow for with pain a higher expression of KORs and MORs in NAc faster and more accurate emotional predictions, and therefore during both abstinence and the reintroduction periods. constitute critical cues for social interactions. However, so far, MORs expression was decreased in VTA and PFC in females studies exploring emotional cross-modal processing in SAUD during the abstinence. Tis efect on MORs expression was relied on static faces, associated with voices from a diferent more pronounced in the presence of pain. Very interestingly individual, largely hampering ecological validity. Besides, in our studies also revealed that these changes in VTA and real life conditions, emotions are often not fully expressed, PFC observed during abstinence are reverted after alcohol so that we have to make guesses based on incomplete infor- relapse, contributing to the understanding of the mechanisms mation. involved in pain induced alcohol relapse-like behaviour in Accordingly, the aim of this study was to assess cross-modal female rats. emotional processing using a new ecological paradigm with Acknowledgements: MINECO Retos de la sociedad dynamic audiovisual stimuli, manipulating the amount of PSI2016-77895-R. emotional information available to the individual. Tirty indi- viduals with SAUD and 30 matched healthy controls perfor- Adipokines (adipocytokines), selected clinical and med an emotional discrimination task requiring to identify nutritional variables in the patients with alcohol emotions (anger, disgust, fear, happiness, sadness) expressed dependence in short movies containing visual, auditory or auditory- Damian Czarnecki1, Marcin Ziółkowski1, Ewa Zekanowska1, visual information of various durations. Te shortest excerpts Barbara Góralczyk1, Jacek Budzynski1, Zofia Rosinska1 revealed the very early emotional sketch (i.e., initial facial (1 Bydgoszcz, Poland) movements and prosody) while the longest ones depicted a more complete emotion. Sensitivity analyses (d’) showed that Introduction: Te hospitalized patients with alcohol de- discrimination levels varied across sensory modalities and pendence frequently have disturbances of nutritional status. emotions, and increased with stimuli duration in both groups. Te problems with the nutrition may be connected with Individuals with SAUD’s performances improved from uni- changes of secretion of adipokines and the clinical status modal to cross-modal conditions, but their discrimination associated with alcoholism. for cross-modal stimuli was impaired for anger and fear. Tis Aim: Te purpose of the study was to assess the adipokines defcit was not infuenced by the amount of information dis- and clinical variables and anthropometric variables in the played, suggesting that it persists even when more emotional patients with alcohol dependence. information is available. Results are discussed in light of the Patients and methods: Te study was conducted among 59 predictive mechanisms underlying emotion recognition, and men hospitalized in the unit of short-term therapy for addic- converge with earlier fndings to ascribe a specifc role for tion. In every subjects were assessed the clinical, anthropome- anger and fear in SAUD. tric variables and the endogenous adipokines. Te laboratory tests was performed using the ELISA. A new animal model of pain-induced alcohol Results: It was shown that the leptin concentration was relapse: Involvement of mu and kappa opioid lower in the patients who started treatment in hospital (with receptors in the mesocorticolimbic system declared monthly abstinence) than in the patients after at 1 1 Javier Cuitavi , Jesús David Lorente , Sandra Fernández- least four weeks of hospitalization. In the patients at the Rodriguez1, Yolanda Campos-Jurado1, Raquel Montón-Molina1, beginning of hospitalization concentration of the apelin was José Luís González-Romero1, Lucía Hipólito1 (1 Burjassot, Spain) lower than in the control group. Te higher concentration of leptin was correlated with the higher BMI (r=0.460) and the Several clinical studies have undercovered that pain may higher %FM (r=0.464). Te lower concentration of visfatin lead to alcohol relapse in patients with a previous history of was correlated with the experiences of alcohol craving (r= alcohol use disorder. Unfortunately, we are still lacking a valid -0.282). Te lower level of %FM, BMI, MAC and WHR was animal model to investigate the underlying neurochemical correlated with higher frequencies of alcohol craving. basis of this efect. According to that an alcohol intermittent Conclusion: Te concentration of adipokines (leptin, apelin) administration animal model in combination with an infam- are changing during hospitalization of patients with alcohol matory pain rat model has been created. dependence. Te adipokines (visfatin, leptin), selected clinical Our model showed that all male rats increased the alcohol and nutritional variables are correlated with each other. It consumption after reintroduction, however in the case of seems that the evaluate of factors such as adipokines, clinical female rats only the ones with infammatory pain increased and nutritional variables may be predictors of recovery of their intake over its baseline. Tat may represent that females patient with addiction. have an increased vulnerability to relapse in the presence of Involvement of neuroinflammation in the effects pain. Besides we measured Mu and Kappa opioid receptors of two ethanol binge episodes during adolescence levels (MORs and KORs) in Ventral Tegmental Area (VTA), Prefrontal Cortex (PFC), Amygdala and Nucleus Accumbens on CA1 hippocampal synaptic plasticity

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 373 Congrès

1 1 1 Chloé Deschamps , Catherine Vilpoux , Mickael Naassila , also of education and Justice, to coordinate the formation of 1 1 Olivier Pierrefiche ( Amiens, France) the diferent professionals and the information of the general public, 2) to facilitate the diagnosis and care of the families in Binge drinking is characterised by an ethanol (EtOH) synergy with regional health networks about perinatality and consumption in a short period of time leading to intoxica- addiction and the new FAS diagnosis center at the University tion, drunkenness, blackouts or even coma. Te binge drin- Hospital, 3) and fnally to promote research with the creation king behaviour is commonly seen in adolescents and can be of a cohort of patients. associated with memory impairment but the neurobiological After 3 years of activity and the training of more than 4,000 mechanisms underlying EtOH-induced memory impairment students and 2,000 professionals, the setting up of question- remain unclear. However, memory impairment induced by naires and standards for professional use, a synergy between intermittent EtOH exposure in young adult rats has been the actors of health, medico-social, social of the National related to neuroinfammation in the hippocampus (Vetreno Education and Justice has been created, allowing the iden- et al., 2015). We previously reported in hippocampus slices tifcation of families and, in connection with the Center of male adolescent rats that 48h after Two Ethanol Binge Diagnosis FASD, the diagnosis of about 150 patients. Tis Episodes (TEBE, EtOH, 3 g/kg, i.p., 9h apart), long-term cohort is a unique source in France of malformative, neuro- depression (LTD) of synaptic transmission – the cellular basis cognitive-behavioral and socio-demographic data. It is based of learning and memory – was abolished and associated with on a biological collection in order to propose an integrative memory impairments. approach of the neurobiological mechanisms as genetics Here we tested the efects of the anti-infammatory agent (presence of genomic variations in 13% of the patients) and minocycline (45 mg/kg, i.p.), administered alone or 30 min epigenetics (search for a specifc methylation profle for early before each EtOH exposure on LTD disruption in hippo- screening). campus slices from male adolescent rats, and we performed immunolabelling for TLR4 after EtOH. We found that Our region has developed a device that responds point by minocycline alone had no efect on LTD while pretreatment point to the recommendations of the new National Action completely prevented LTD abolition 48h after two binge epi- Plan against Addictions MILDECA 2018-2022. It could be sodes. In parallel, TLR4 expression was increased at that time a model for setting up other centers, both in France and in point after EtOH. Our study demonstrated that synaptic other overseas regions. plasticity impairment induced by two EtOH binge episodes Suppression of ethanol induced during adolescence involves neuroinfammatory mechanisms. neuroinflammation by PPAR- agonists in an The FASD Resource Center in Reunion Island: Back animal model of fasd to 3 years of activity P.D. Drew1, J.W. Johnson1, T. Rafferty1, C.J.M. Kane1 (1 Little Bérénice Doray1, Barbara Delmotte1, Karine Josse1, Stéphanie Rock, USA) Sotaca1, Thierry Bafinal1, Stéphanie Robin1, Justine Lanneaux1, Augustin Rousselle1, Marilyn Tallot1, Marie-Line Jacquemont1, Maternal alcohol consumption can lead to developmental Alizé Payet1, Lucie Rebourg1, Sonia Henkous1, Nathalie Penard1, maladies associated with Fetal Alcohol Spectrum Disorders Marine Gayet1, Agnès Cudenet1, Michel Spodenkiewicz1 (FASD). FASD is a leading cause of mental retardation and is (1 La Réunion, France) associated with substantial lifetime disabilities. Unfortunately, there is no efective pharmaceutical treatment. Tus, the Concerning 1 per 100 births, Fetal Alcohol Spectrum need for new therapeutic strategies to mitigate the conse- Disorders (FASDs) constitute a major but preventable quences of FASD is of great importance. Using our third cause of neurocognitive disorders and social maladjustment. trimester-equivalent mouse model of FASD in which mice Nevertheless, screening, diagnosis and management of are treated with 4 g/kg ethanol per day via intra-esophageal patients and families remain difcult, due to a lack of know- gavage on postnatal days 4-9, we showed that ethanol pro- ledge of this condition by the various professionals concerned duces prevalent neuronal and glial cell loss in the developing but also difculties of access to care of these patients. brain. Further, surviving microglia undergo a morphological With the highest rate of FASDs in France (Public Health change to an activated pro-infammatory phenotype. Tis France, 2018), Reunion Island was selected in 2015 as a is accompanied by an increase in expression of cytokines pilot region for prevention, screening, diagnosis and care and chemokines associated with neurodegeneration, neuro- for FASDs (Interministerial Mission against Drugs and infammation, and neuropathology. We further demonstrated Addictive behavior MILDECA 2013-2017 plan). Te FASD that the FDA-approved PPAR- agonist pioglitazone can Resource of Reunion Island constitutes the central link of attenuate ethanol-induced cellular toxicity, microglial mor- this experimental Action Plan against FASD. It is funded by phological change, and expression of infammatory molecules. the Regional Health Agency of Indian Ocean (ARS-OI) and Tis suggests that PPAR- agonists may hold therapeutic MILDECA and managed by the medicosocial Foundation potential for those afected by FASD. We also evaluated the “Père Favron” in partnership with the University Hospital. efect of the PPAR- agonist docosahexaenoic acid (DHA) Its missions are multiple : 1) to identify and put together the on ethanol-induced neuroinfammation. DHA is an -3 diferent actors of health, medico-social and social sectors, but fatty acid that possesses anti-infammatory activity, and is

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 374 Congrès abundant in the brain. It is available in dietary sources, fsh incidence of alcohol abuse in Belgium is 10%. In another oil, and commercial baby formula. We treated postnatal mice Belgian survey in 2013, it is mentioned that 90% of alcohol with DHA 1-2 hours prior to ethanol treatment. Brain tissue abusers do not get specialized help. However, it is known that was harvested on postnatal day 10 and gene expression was early intervention and ease of access to specialized care gives quantifed. DHA suppressed ethanol induced expression of a better prognosis for people with alcohol dependence issues. pro-infammatory molecules including the cytokines IL-1 Tis is also the clinical observation shared by our team, the and TNF- in the brain. Tus, PPAR- agonists including alcohol rehabilitation center in CHU Brugmann (Unit 72). DHA and pioglitazone are identifying new mechanisms of Hospitalized patients for withdrawal frequently arrive with alcohol-induced brain pathogenesis. Further study is needed few therapeutic levers. For example, their family is often to evaluate their safe and efective use for treatment of the exhausted by their long course of alcohol dependence. Patient neuropathological consequences of FASD. resources are diminished in fnancial, social, professional and Supported by NIH AA026665, AA024695, AA027111. cognitive terms. And the negative impact of alcohol abuse on neurocognitive functions does not facilitate the patients Alcohol-related abnormalities in the early ability to combat addiction. postnatal period can be corrected by multitarget Description of our program: We have created a two-day low-affinity agonist of sigma-1, MT1 and MT3 program whose goal is to inform participants and assess receptors both their physical and psychological health (in relation to 1 1 Andrei Durnev (Moscow, Russia ) alcohol use). Our program is open to anyone concerned about his drinking behavior. Our primary objectives are to inform Alcohol intake leads to negative reproductive outcomes in persons who are ambivalent about their alcohol consumption men and complications in pregnancy followed by behavioral and to refer them if necessary to the appropriate services that and physiological alteration in ofspring. Previous studies could provide them with adapted assistance. Our aim is also showed the ability of (Afobazole®), a low- preventing the development or aggravation of dependence afnity agonist of sigma-1, MT1 and MT3 receptors, deve- and to favor intervention as early as possible with an acces- loped for treatment generalized anxiety disorder, to prevent sible and attractive program. ethanol-induced DNA damage in embryonic cells and fetal Our program consists of an evaluation of alcohol consump- abnormalities. Te aim of the work was to evaluate possibility tion via a psycho-medical assessment, providing clear infor- of pharmacological treatment of ethanol-induced early post- mation, while being attentive to the persons. We want to natal disorders in outbred rat ofspring from chronic ethanol- provide participants with a complete assessment of their exposed male rats (CEE) or prenatal ethanol-exposed female consumption. To do this, our method of communication and rats (PEE). animation are based on Motivational Interviewing. We em- In CEE model male rats had 10% v/v ethanol as the only phasize the importance of follow-up afterwards, our program source of liquid for 24 weeks. Sperm morphology was exami- has the objective to motivate and to incentivize the setup of ned in stained slides (100×magnifcation, 200 sperms/rat). In a personalized care or prevention project. PEE model dams had ethanol (4.3 g/kg/day, 40% v/v, orally) We hope to be able to answer any questions clearly and to from 10th to 19th day of pregnancy and were pretreated with allow participants to discuss and think about their alcohol fabomotizole (1-10 mg/kg, orally, daily) 15 min prior to etha- use in complete confdentiality and without any judgment. nol. Newborns from rats after CEE and PEE were evaluated for unconditional refexes formation (“turning on the plane” The role of alcohol on iron metabolism and and “avoiding the edge” tests) and muscle tone (“horizontal erythropoiesis in acute and chronic alcohol mouse rope” test) on 5th day of life. model CEE model resulted in signifcant increase of sperm abnor- Faiza Faraz1, Teresa Peccerella1, Sebastian Mueller1, malities, however no changes in sensory-motor refexes and Marina Scheller-Wendorff1, Vanessa Rausch1 (1 Heidelberg, muscle strength in ofspring were revealed. In PEE newborns Germany) the main indices of refexes and muscle tone were reduced by 1,5-2 times. Fabomotizole at anxiolytic doses prevented alcohol-induced neurodevelopmental damage. Background and aims: Chronic alcohol consumption leads to Tus, early postnatal abnormalities in rats exposed to ethanol multiple illnesses as well as to deleterious efects on hema- in utero can be corrected by fabomotizole perhaps due to topoiesis. However, little is known about alcohol efect on cytoprotective, neuroprotective and antioxidative properties. erythropoiesis. Our aim was to investigate the relationship between alcohol consumption, iron overload and erythro- “Alcohol and you?” A two day assessment poiesis. program with a multidisciplinary team Methods: Te efect of ethanol ingestion on erythropiesis L. Fabry1, A. Rogiers1, C. Hanak1 (1 Bruxelles, Belgique) was determined in male C57BL/6 wild-type mice (8 weeks old) treated with 2 gavages of alcohol 31.5 v/v (acute group), Introduction: Te prevalence of alcohol use disorders in 20% alcohol in drinking water for 4 weeks (chronic group) Europe is 7,5% according to WHO data in 2010. Based on and control group was given normal drinking water followed the health survey of L. Gisle and collaborators in 2013, the by 2 gavages of maltodextrin (45% w/v). At the end animals

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 375 Congrès were sacrifced and peripheral blood, spleen, kidney, liver, measures at 6‐month post treatment follow‐up. and bone marrow were collected. Erythroid diferentiation Conclusions: Supportive text messages provide an early initial and erythroid maturation was analyzed by fow cytometry. beneft in decreasing symptoms of depression and stress, with Hepcidin, SMAD6, SMAD7, and HO-1 mRNA levels from a further positive impact on alcohol consumption following a liver and spleen were assessed by qRT-PCR and STAT3 and longer treatment period. Benefts did not persist six months ferroportin by western blot. Parafn embedded sections were after the intervention ended. also histoligically analyzed. Results: We observed reduced numbers of RBCs along Historical keypoints in the concept, definition and with reduced cellularity in bone marrow, splenomegaly and pathogenesis of alcoholic cardiomyopathy increased liver weight in both short and long term alcohol J. Fernández-Solà1, M. Guitart-Mampel1, G. Ferrer-Curriu1, mouse models. Number of megakaryocyte-erythroid pro- E. Tobias-Baraja1, G. Garrabou-Tornos1, A. Planavila1 gentors (MEPs) was drastically reduced in acute group sug- (1 Barcelona, Spain) gesting impaired early stages of erythropoiesis. However, in chronic ethanol exposure a high number of proeryhtroblast Background: Alcoholic Cardiomyopathy (ACM) is at present (Ter119neg CD71high) and low number of late erythroblasts a well defned clinical and pathological entity. However, over (Ter119high CD71med) was detected. Acute as well as chro- the years many previous doubts have emerged concerning to nic alcohol exposure led to signifcant hepcidin suppression its existence, defnition and physiopathological concept. accompnied by suppression of SMAD6 and 7 mRNA and Aim: 1) To review historical key points in the establishment an induction in HO-1 levels suggesting heme degradation. of concept and defnition of ACM. 2) On this basis, to per- Ballooned hepatocytes and a large number of eryhthrocytes form a future overview projection on how to prevent future were observed in liver during histological analysis. ACM in chronic consumers Conclusion and outlook: Hematopoietic tissues displayed a Methods: We analyze 12 diferent critical historic points dramatic increase in early erythroblast numbers, but these on the scientifc knowledge on ACM. 1) Hippocrates’ reco- fail to diferentiate. Tis was accompanied by disturbances in gnition. 2) First clinical modern descriptions. 3) Ethanol systemic iron homeostasis mediated by hepcidin suppression itself or ethanol contaminants cause ACM? 4) Te nutri- in the liver. tional hypothesis. 5) Ethanol or acetaldehyde? 6) Te dose- dependent relationship between alcohol and heart function. Alcohol use disorder and comorbid depression: 7) ACM in women. Te same as men? 8) ACM and control A randomised controlled trial investigating the drinking. 9) Te multisite pathogenic hypothesis. 10) Heart effectiveness of supportive text messages in Remodeling in ACM. 11) Te heart’s secretor role: alcohol aiding recovery and cardiomyokines. 12) How to prevent future ACM in C.K. Farren1, H.O. Reilly1, A. Hagerty1, A. Farrell1, chronic consumers? D. MacLoughlin1 (1 Dublin, Ireland) Conclusions: After the Hippocrates’ defnition of alcoholic cardiomyopathy, its modern clinical recognition delayed Aim: Te aim of this randomised controlled trial was to exa- more than one millennium. Teir pathogenic bases just have mine the impact of daily supportive text messages over a six‐ emerged 100 years ago and are still on construction. Possible month treatment period on mood and alcohol consumption efective pathogenic intervention are just planned for nearly in individuals with a dual diagnosis of alcohol use disorder future. (AUD) and depression following completion of an inpatient With support of Grants from La Marato 2015 33 30/31 and treatment programme. CIBEROBN. Method: 95 adult participants with AUD and comorbid depression were recruited. Te intervention group (n=47) N-acetylcysteine alters GLT-1 and Fos B received twice‐daily supportive text messages over 6‐months expression in the dorsolateral striatum of while control participants (n=48) had treatment as usual for long-term ethanol-experienced rats during the a 6‐month period, with an added 6‐month post‐treatment abstinence period follow‐up for both groups. Drinking history in the previous S. Fernández-Rodríguez1, C. Espósito1, L. Granero1, A. Polache1, 90 days as well as symptoms of depression, anxiety and stress M.J. Cano-Cebrián1, T. Zornoza1 (1 Valencia, Spain) were measured at baseline, 3‐ and 6‐month treatment points and 6‐month post treatment follow up. Alcohol Use Disorder (AUD) is a chronic and recidivant Results: Depression scores (p=0.02) and perceived stress neurobehavioural disorder which supposes a serious health scores (p<.01) were signifcantly reduced at 3‐month treat- as well as economic problem worldwide. Nowadays, relapse ment point in the intervention group relative to control prevention is considered the main target for therapies against participants with small to medium efect. Te intervention drug addiction, but after decades of research, few drugs have group also showed a signifcantly greater reduction in units been marketed for this purpose. In the last decades, dere- per drinking day from baseline to 6‐month treatment point gulation of glutamate homeostasis has been postulated as compared to the control group with a medium efect size one of the critical points in cue-induced relapse. In previous (p=0.03). Tere were no diferences in drinking or mood research, our laboratory evidenced that N-acetylcysteine

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 376 Congrès

(NAC), a safe and well-tolerated marketed drug, is able to Perceptions of alcohol use during pregnancy in block the Alcohol Deprivation Efect (ADE) in long-term France, Spain and Portugal – A cross-cultural ethanol-experienced rats, but the mechanism underlying qualitative study its anti-relapse efcacy is complex and still remains unclear. Renata Franco1, Belén Charro2, María Raul Xavier3 (1 , We hypothesized that the anti-relapse efect displayed by France, 2 Madrid, Spain, 3 Porto, Portugal) NAC could be related to a normalization of glutamatergic adaptations triggered by continuous ethanol experience. In Background and aims: Considering children prenatally the present research, we explored the expression of glutamate exposed to alcohol present substantial challenge to parents, type 1 transporter (GLT-1) and ∆Fos B (a transcription schools, and societies and considering minimum safe dose of factor that is related to the mechanisms by which addictive alcohol during pregnancy is unknown, WHO, EU and dif- drugs produce stable changes in the brain) in the dorsolateral ferent countries suggest zero consumption. Despite, research striatum, a region implicated in the addiction process through shows that there is a substantial number of women who the control of habit formation. During the fourth abstinence continue to drink. period, 30 male Wistar rats were subcutaneously adminis- Taking into consideration that information is needed to make tered 0, 60 or 100 mg/kg NAC once daily during 14 days. an informed decision about alcohol use during pregnancy, Animals were sacrifced before ethanol reintroduction and understanding the accessibility and quality of information Western Blot analysis was performed. Te obtained results available to pregnant women is an issue for research. Tis may suggest a plausible mechanism for NAC previously work presents a qualitative study exploring attitudes of demonstrated anti-relapse efcacy. Portuguese, Spain and French pregnant women regarding alcohol use during pregnancy, knowledge about the impact HCC alcohol exposure inhibits the suppressor of of alcohol use during pregnancy, accessibility and quality of tumor SLAMF3 information available. Grégory Fouquet1, Constance Marié1, Charles-Antoine Papillon1, Methods: Semi-structured interviews were conducted with Éric Nguyen-Khac1, Mickael Naassila1, Hicham Bouhlal1, 20 French (Toulouse), 19 Portuguese (Porto) 30 Spanish Ingrid Marcq1 (1 Amiens, France) (Madrid) pregnant women. Interviews were audio recorded and transcribed verbatim. Data were qualitative analyzed using a semi-inductive approach. Teoretical saturation was Background and Aims: HepatoCellular Carcinoma (HCC) achieved in both groups. is one of the most frequent cancer worldwide and the fourth Results: Six of the twenty French, six of the nineteen one in cancer-related deaths. One of the main important Portuguese and nine Spanish pregnant women reveals to etiology of HCC is chronic alcohol consumption. A study drink at some point during pregnancy: in both countries from Costentin et al. described a decrease of global survival during festive events. Pregnant women (French, Portuguese in alcoholic HCC patients compared to other etiologies. and Spanish) described mixed messages and confusions about Interestingly, we identified in our laboratory, a receptor, consequences of alcohol consumption during pregnancy. In SLAMF3 at the surface of hepatocytes. Te expression of Portugal, participants reported several limitations concerning SLAMF3, a member of Signalling Lymphocytic Activation accessibility and quality of information available for pregnant Molecules family, is lost in cancerous hepatocytes compared women and social pressure to drink in festive occasions. to healthy cells. SLAMF3 overexpression in HCC cells French and Spanish participants argued that it is easy to fnd induces tumors regression in a xenograft model, which was information related to alcohol and pregnancy. explained in part, by the decrease of MAPK pathway activity. Furthermore, alcohol consumption is known to induce the A multi-level approach for prevention of underage MAPK pathway activation. In this context, we investigated drinking on california indian reservations: Efficacy the efects of alcohol exposure on the tumor suppressor efect and mechanism of behavior change in adults and of SLAMF3 and signalling pathways implicated in these adolescents mechanisms. D. Gilder1, J. Geisler1, J. Luna1, D. Calac1, J. Lee1, R. Moore1, Method: HCC cell lines were exposed to increased concen- C. Ehlers1 (1 La Jolla, USA) trations of alcohol (0 to 160mM). Effect of alcohol on SLAMF3 expression was analyzed by flow cytometry. Purpose: Te individual and community level interventions SLAMF3 expression was also studied by RTqPCR in HCC of a successful multi-level prevention of underage drinking patients from diferent etiologies. program were assessed for efcacy and mechanism of beha- Results: We showed that alcohol exposure decreased vior change in a community sample of reservation dwelling SLAMF3 expression. Simultaneously, we observed a signi- Southern California American Indian adults and adolescents. ficant decrease of SLAMF3 expression by RTqPCR in Methods: Te four interventions were: (1) youth participatory alcoholic patients compared to patients with other etiologies. community mobilization to develop a billboard message, Conclusion: We revealed the involvement of alcohol in the (2) convenience store checks to discourage alcohol sales to loss of expression of the tumor suppressor SLAMF3 in minors, (3) Motivational Interviewing (MI) vs. Psycho- HCC. Tis observation might explain the aggressiveness of education (PE) in 112 youth, and (4) 298 community alcoholic HCC compared to others etiologies. education and outreach events to adults and youth aimed

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 377 Congrès at preventing underage drinking. One hundred and twenty consumption of tobacco or liver function impairment, but adults and 100 youth were surveyed. Frequency analysis, with age over the median (LR=5.94; p=0.015). In addition, Wilcoxon signed ranks tests, and logistic regression were used altered body composition (assessed in 313 patients by total to analyze survey results. body densitometry) was related with the time at which neo- Data and results: Adults (awareness of the intervention, took plasia developed, so that patients with trunk fat below the action to reduce teen drinking as a result of that awareness): median (LR=3.59; p=0.058; B=4.06; p=0.044), total BMD Billboard (49%, 75%), Store Checks (77%, 75%), MI vs. over the median (LR=3.32, p=0.066; Breslow=4.22; p=0.04), PE (63%, 80%), Outreach (65%, 76%). Relative strengths or lumbar t-score over the median (LR=6.31; p=0.012; of interventions (z score, p-value): Outreach > Billboard Breslow=6.93; p=0.008) developed cancer earlier, especially (2.52, 0.012), Outreach > MI vs. PE (2.70, 0.007), no among the 149 cirrhotic (LR=7.16; Breslow=9.71; p<0,001). other diferences. Mechanism of behavior change associated Tese results are similar to those observed among women with the overall program (OR, p-value): movement from with breast cancer, but to our knowledge, they have not been Precontemplation to Contemplation with awareness of pro- reported among alcoholics. blem (6.04, 0.001), from Precontemplation to Contemplation with considering change (1.94, NS), from Contemplation to Neural responses to multisensory alcohol cues in Preparation (10.80, 0.001). Teens (awareness of the interven- heavy-drinking smokers tion, reduced drinking as a result of that awareness, doesn’t Kimberly Goodyear1, Robert M. Swift1, Lorenzo Leggio1, drink): Billboard (31%, 30%, 61%), Store Checks (68%, 32%, Carolina L. Haass-Koffler1 (1 Providence, USA) 52%), MI vs. PE (61%, 32%, 52%), Outreach (74%, 29%, 58%). Background: Approximately 80% of individuals with an Relative strengths of interventions (z score, p-value): alcohol use disorder (AUD) are also cigarette smokers, and Outreach > Billboard (2.11, 0.035), no other diferences. despite previous research on functional magnetic resonance Mechanism of behavior change associated with the overall imaging (fMRI) cue-reactivity, the behavioral and neural program (OR, p-value): movement from Precontemplation responses to alcohol cues in heavy-drinking smokers have to Contemplation with awareness of problem (18.1, 0.006), not been investigated. from Precontemplation to Contemplation with considering Study objectives: Te goal of this pilot study was to examine change (9.5, 0.027), and from Contemplation to Preparation the efects of visual and olfactory alcohol cues on blood- (9.5, 0.027). oxygen-level-dependent (BOLD) activity in heavy-drinkers Conclusions: All four interventions were associated with during fMRI scan. high levels of awareness and intervention to reduce teen Methods: Heavy-drinking smokers (n=10) participated in the drinking (adults) and reducing drinking (teens). With some alcohol fMRI cue-reactivity task. We implemented an alcohol exceptions, perceived strengths of interventions were similar. cue-reactivity task, where participants, after being exposed For both adults and teens, interventions were associated to alcohol and neutral cues (visual and olfactory), rated their with movement along the transtheoretical model of stages of craving for alcohol and cigarettes with visual analog scales. behavior change. Independent samples t-tests were implemented to compare Support: National Institute on Alcohol Abuse and Alcoholism alcohol and cigarette craving during alcohol and neutral cues. (NIAAA) grant R01 AA023755. Further, whole-brain and region of interest (ROI) analyses were done to compare BOLD responses to alcohol and neu- Incident neoplasia among heavy alcoholics: tral cues. Lastly, correlation analysis was done on activation Relationship with body composition in ROIs and baseline craving and drinking and smoking Lourdes González-Navarrete1, Iván Ribot-Hernández1, behaviors. Víctor Vera-Delgado1, Candelaria Martín-González1, Mª José Results: Our behavioral results showed that participants had Sánchez-Pérez1, Julio Alvisa-Negrín1, Antonio Martínez-Riera1, higher alcohol craving during alcohol cues compared to neu- Emilio González-Reimers1 (1 Tenerife, Canary Islands, Spain) tral cues (p<.05). Further, our whole-brain analysis revealed signifcant activation in the right lingual gyrus (p<.005). In a series of 408 alcoholic patients consecutively admitted to Te ROI analysis showed signifcant activation in the right the hospitalization unit of the Internal Medicine Service and orbitofrontal cortex (OFC) (p<.05) when comparing alcohol followed up for 12 years we observed that the proportion neo- to neutral cues. Correlation analysis indicated that there plasia among cirrhotic and non-cirrhotic was similar (21.20% was a positive associations with baseline alcohol craving and among cirrhotic and 21.43% among non-cirrhotic (2=0). activation in the right ventral striatum (VS) (p<.05) and the Most cancer afected the oropharyngeal area (28.57%), colon left anterior cingulate cortex (ACC) (p<.05). Tere were also (20.24%) and prostate (13.10%). Fourteen patients developed positive associations with total alcohol drink in the ninety a multiple neoplasia. Of the 87 patients with cancer, 14 were days prior to the experiment and activation in the right VS already diagnosed with a neoplasm when they entered the (p<0.0001), left VS (p<.01) and left ACC (p<.0001). study, whereas 73 developed an incidental neoplasia along Conclusions: We have provided preliminary evidence that the study period. Te incidence of neoplasia was not related there are distinct behavioral and neural patterns in response to ethanol consumption, the presence of liver cirrhosis, the to alcohol cues in heavy-drinking smokers.

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 378 Congrès

A polymer- conjugate ameliorates the performed. For evaluation of the efects of EOS in ethanol neuroinflammation associated with chronic alcohol abstinence syndrome, healthy Wistar rats were enabled to treatment in mice voluntary drinking of 4.5%, 7.5% and 9% v/v alcohol for C.M. Cuesta1, F. Ibañez1, R. Lopez-Hidalgo1, J. Urena1, ffteen days. Te behavior studies were conducted employing A. Duro-Castaño1, A. Armiñán1, M.J. Vicent1, M. Pascual1, tail suspension test and forced swim test on day 16th, 17th C. Guerri1 (1 Valencia, Spain) & 18th and peak ethanol withdrawal syndrome was deter- mined. EOS (100, 300, and 500 mg/kg) and standard drug Several evidences demonstrated that alcohol, by activating fuoxetine were administered orally during withdrawal symp- the brain immune receptors TLRs and NLRs, can induce toms. Alcohol biomarkers like ALT, AST, ALP, GGT, and infammatory mediators and cytokines/chemokines trigge- MCV were estimated by using commercially available kits. ring neuroinfammation and brain damage. Terefore, it is concentrations were measured in the plasma, hip- important to develop efective therapies to reduce or ame- pocampus and prefrontal cortex using the rat ELISA kit. Te liorate the neuroimmune system activation. Considering that gene expression analysis of GRIN1, GRIN2A, and GRIN2B curcumin has important anti-infammatory and antioxidant of N-methyl-D-aspartate receptors (NMDAR) subunits in properties, but low , we have used a polymer- the hippocampus and the prefrontal cortex were also exa- curcumin conjugate (PCC) derivatised to be able to cross the mined by RTqPCR. Te results displayed that no observed blood-brain barrier through the LRP-1 receptor in order to adverse efect level (NOAEL) for EOS was higher than block neuroinfammation. Te conjugation of curcumin to a 2000 mg/kg, orally. Te deregulated levels of alcohol markers biodegradable polymeric carrier enhances curcumin efciency and serotonin following ethanol abstinence in the plasma, and controls drug release by the presence of bioresponsive hippocampus, and prefrontal cortex were also reversed by polymer-drug linkers (pH-labile esters). We used glial cells EOS at doses 300 and 500 mg/kg. EOS exerted a signifcant in culture incubated with and without ethanol and in the protective efect at doses 300 and 500 mg/kg, but 100 mg/kg presence or absence of PCC. For the in vivo experiments, showed insignifcant protection against alcohol abstinence- mice treated with or without ethanol during two months induced depression like behavior in both FST and TST. were administered PCC intravenously, two times/week. In Te increased expression levels of GRIN2A and GRIN2B vitro results experiments demonstrated that PCC is not following ethanol abstinence were also reversed with a higher toxic for glial cells and protects against ethanol-induced cell dose of EOS (500 mg/kg) treatment. Tus, the results of the toxicity. Our in vivo result shown that PCC administration study reveal that EOS has a remarkable protective role in protects ethanol-induced the up-regulation of infammatory the ethanol abstinence-induced depression by modulating mediators (TLR4, iNOS, COX-2, IL-1 , frantalkine), in alcohol markers, serotonin levels, and expression of GRIN2A, prefrontal cortex and in medial cortex of chronic ethanol GRIN2B gene of NMDAR signaling in rats. mice. We further observed that chronic ethanol-treatment signifcantly up-regulated some miRNAs (miRs 146a-5p and Nucleus reuniens of ventral midline thalamus is let-7b-5p) that modulate neuroinfammation in the medial highly susceptible to permanent neuron loss in the cortex. PCC administration supresses’ ethanol-induced rat model of binge drinking during third trimester changes in these miRNAs. In summary, our results support Z.H. Gursky1, A.Y. Klintsova1 (1 Newark, USA) the benefcial efects of PCC administration by attenuating the neuroinfammation associated with chronic alcohol abuse. Supported by SAF2015-69187R. Individuals with fetal alcohol spectrum disorders often have difficulty performing high-demand cognitive tasks (i.e., Ocimum sanctum suppresses alcohol abstinence- have impaired “executive function”). Executive function is induced depression-like behavior through supported by communication between 2 parts of the brain: regulation of biochemical and GRIN2A, GRIN2B hippocampus and prefrontal cortex. We hypothesized that gene expression of NMDA receptor signaling in rats midline thalamic nucleus reuniens (responsible for coordi- Lal Gupta Girdhari1, Sharma Anamika2 (1 Mumbai, nating activity between prefrontal cortex and hippocampus) Maharashtra, India, 2 Solan, Himachal Pradesh, India) could be afected by developmental alcohol exposure (AE). We use 3 AE paradigms in Long Evans rat during a period India’s Queen of herbs Tulsi (Ocimum sanctum Linn., family comparable to human third trimester to demonstrate the Labiatae) have huge medicinal uses and traditionally being mechanism by which AE results in short- and long-term used for the treatment of alcohol disorders. However, its neuroanatomical damage within nucleus reuniens, and what underlying mechanism(s) of action have not been adequa- cell types are most vulnerable. tely addressed. Terefore, we evaluated the efect of Ocimum Te frst paradigm, high-dose (5.25 g/kg/day) AE on single sanctum in alcohol abstinence-induced depression, developed postnatal day (PD) 7 demonstrated that alcohol-induced cell following long-term voluntary alcohol intake in rats. Te loss in reuniens in adulthood is caused by alcohol-induced hydro-alcoholic extract of Ocimum sanctum leaves (EOS) was cell death in males and females. frst characterised for the presence of oleanolic acid (0.54% Additionally, AE reduced reuniens volume. The second w/w), eugenol (0.39% w/w) and caryophyllene (0.02% w/w) paradigm, high-dose AE on PD4-9 caused a selective loss and subsequently acute, sub-acute toxicity studies were also of neurons, but not non-neurons, reducing neuron-glia ratio,

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 379 Congrès as well as a reduction in volume of reuniens. No alterations intake, the Ai activation decreased it. Tis suggest that Ai were observed in the neighboring rhomboid nucleus. Te may partly provide the Acb with the excitatory input enhan- third paradigm examined the impact of high-dose (5.25 g/kg/ cing alcohol drinking, whereas the efects of Ai stimulation day) and moderate-dose (3.00 g/kg/day) AE on reuniens. We are mediated by other, still unknown circuits. observed signifcant neuron loss in both sexes at both doses, but volume was only reduced following high-dose AE (repli- (±)–Baclofen in alcohol use disorder: Identification cating and extending fndings from both prior paradigms). of responders and of the role of dopamine release Neither dose of alcohol altered the number of microglia in in the nucleus accumbens in the efficacy of the reuniens. Taken together, these experiments indicate that different enantiomers reuniens is highly susceptible to damage, over various levels Jérôme Jeanblanc1, Victor Echeverry-Alzate1, Pierre Sauton1, of drinking, even following brief exposure. Virginie Jeanblanc1, Mickael Naassila1 (1 Amiens, France)

Chemogenetic manipulation of nucleus accumbens Studies to evaluate the efcacy of (±)-baclofen (the racemic and insula activity modulates alcohol consumption R(+) and S(-) form) in the treatment of alcohol dependence in rats have yielded mixed results and lively debate about the bene- 1 1 2 1 Mia Haaranen , Annika Schäfer , Sara Häkli , Petri Hyytiä ft/risk ratio at the international level. Recent studies have (1 Helsinki, Finland, 2 Joensuu, Kuopio, Finland) suggested that diferent enantiomers may help to explain, at least in part, the contrasting results and the great variability in Functional brain imaging in humans and rodents has impli- treatment response. We investigated the efectiveness of each cated two brain regions in the development and maintenance of the enantiomers on self-administration of alcohol in either of alcohol use disorders (AUDs), the nucleus accumbens alcohol-dependent rats or binge drinker rats. We have shown (Acb) and the anterior insula (Ai). Both structures are part of that the R(+) form is more efective in reducing alcohol the mesocorticolimbic reward system: Te Acb is located in consumption, craving and relapse than the racemic form and the striatum and has been associated with mediating emotio- at a lower dose (1.5 mg/kg vs. 2 mg/kg). Almost 30% of rats nally rewarding sensations elicited by natural rewards as well signifcantly increased their alcohol consumption (+50%) af- as drugs of abuse. Te Ai is a cortical structure implicated ter the administration of either the racemic or the S(-) forms in the generation of interoceptive cues and decision making of baclofen. R(+)-baclofen only leads to a sharp decrease during goal-directed actions. in alcohol consumption in both rat populations. We also To characterize the functional role of Acb and Ai in the regu- found that the racemic and the R(+)-baclofen are both more lation of voluntary alcohol consumption, we used chemoge- efcient in males than in females rats. Finally, using the fast netic manipulation of neuronal activity through designer re- cyclic voltammetry technic on nucleus accumbens containing ceptors exclusively activated by designer drugs (DREADDs). brain slices, we found that the racemic and the R(+)-baclofen Te viral construct carrying either an excitatory (G(q)-) or reduced the DA release whereas the S(-)-baclofen increases inhibitory (G(i)-) DREADD was injected bilaterally into the release of dopamine. Terefore, the S(-)-baclofen seems the Acb or Ai of alcohol-preferring AA (Alko, Alcohol) rats to induce opposite efects both on the behavior and the trained to voluntarily self-administer alcohol (intermittent dopaminergic release than the R(+)-baclofen and thus, the drinking paradigm, 2-bottle choice test, 2 h access to 10% R(+)-baclofen should be a better medication in order to treat EtOH qad). After a four week expression time, DREADDs AUD than the currently prescribed racemic form. were activated by clozapine-N-oxide application (CNO, 10 mg/kg, ip). Neuronal Acb activation resulted in an increase Correlation between self-reported alcohol-intake in alcohol consumption while neuronal silencing lead to a (AUDIT-C) and PEth-concentrations in somatic decrease of alcohol intake. Neuronal Ai stimulation produced patients admitted to hospitals in Oslo and a decrease in alcohol drinking. Neuronal silencing did not Moscow show a change in drinking habits. Water consumption was Benedicte Joergenrud1 (1 Oslo, Norway) not afected in either of the groups. As the Ai has direct aferent connections to the Acb, we Background: AUDIT-C has traditionally been one of the decided to additionally examine the efect of pathway speci- most commonly used screening tools for identifcation of fc manipulation targeting the connections originating from harmful alcohol use. In recent times, the use of the biomarker the Ai and terminating in the Acb. Here, Cre-dependent phosphatidylethanol (PEth) (16:1/18:0) has been applied in DREADDs were injected into the Ai while the correspon- several clinical settings for detecting harmful alcohol use, as ding Cre-factor was applied to the Acb. Te results showed a it corresponds directly with alcohol consumption. However, statistically signifcant increase in alcohol consumption after there have been few studies that investigate the relationship CNO administration in the excitatory G(q) group while between AUDIT-C and PEth-concentrations. In our study drinking in the G(i) group remained unaltered. we wanted to see the correlation between PEth concentra- The results presented here show that both Acb and Ai tions and self-reported alcohol-consumption during the last contribute to voluntary alcohol consumption. However, while 12 months. activation of Acb or the Ai->Acb projection increased alcohol Methods: AUDIT-C data and PEth concentrations was

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 380 Congrès collected from 1897 Norwegian and Russian somatic patients lowing alcohol exposure and variants in this gene segregating during a period from 11/2016 to 12/2017. Te AUDIT-C among the BXD population may modulate alcohol intake in data was converted to weekly grams of alcohol consumption response to stress. by multiplying drinking events (AUDIT item 1) with the Conclusions. Tese results are the frst to show a genetic basis average number of alcoholic units consumed in a normal for individual diferences in the efects of chronic stress on drinking event (AUDIT item 2), and adding the alcoholic alcohol consumption and nominated a likely candidate gene. units from binge drinking (AUDIT item 3). We subsequently Future work will involve validating Nos1 and discovering correlated the weekly consumed alcohol with PEth concen- other genes underlying stress-related alcohol consumption trations. in humans. Results: When dividing the patient-population by country and gender we found that most patients drink from 12.8- Changes in the metabolome of human post-mortem 99.9 grams of alcohol per week. We also found that men brain samples associated with excessive alcohol use drink proportionally more alcohol compared to women on a Olli Kärkkäinen1, Kati Hanhineva1, Pekka J. Karhunen2,3, weekly basis. Mean and interquartile range of PEth increased Jari Tiihonen4,5, Eloise Kok2,3 (1 Joensuu, Kuopio, Finland, with higher self-reported alcohol consumption during the last 2 Tampere, Finland, 3 Fimlab Laboratories Ltd, Finland, 4 Kuopio, 12 months, and a medium correlation efect size was found. Finland, 5 Stockholm, Sweden) Discussion: Converting AUDIT-C scores into alcohol consumption in grams per week makes for a more practical Aims: Alcohol exposure has been shown to alter metabolite and feasible approach in correlating with PEth concentrations. levels in the brain in rodents. Here our aim was to investigate Our study showed that self-reported grams of alcohol consu- if the brain metabolome of humans is altered in association med each week correlated well with PEth concentrations with excessive alcohol use. within two diferent hospital populations. However, a major Methods: We analyzed frozen human post-mortem frontal limitation is that the patients self-reported alcohol consump- cortex samples from persons with history of excessive alco- tion referred to the last 12 months, while PEth concen- hol use (n=97) and controls (n=107). We used non-targeted trations refects alcohol use only during the last 4 weeks. liquid chromatography mass spectrometry method for the metabolomics analyses. Effect of chronic stress on alcohol consumption is Results: We observed diferences between the study groups mediated by genetics in BXD recombinant inbred in the metabolite levels in the post-mortem frontal cortex mice samples. For example, we observed decreased levels of acetyl- 1 1 1 1 Byron C. Jones , Megan K. Mulligan , Lu Lu , Wenyuan Zhao , choline (p<0.001) and GABA (p<0.001) in the alcohol group Robert W. Williams1, Sonia A. Cavigelli2, Elena Terenina3, when compared to the controls, indicating alterations in the Pierre Mormède3 (1 Memphis, USA, 2 University Park, USA, neurotransmitter metabolism. Moreover, we observed in- 3 Castanet-Tolosan, France) creased levels of S-adenosyl-L-methionine (SAM, p<0.001) in the alcohol group when compared to the controls, indica- Background. Te efect of stress on alcohol consumption in ting alterations in the methylation processes since SAM is humans is highly variable and underlying processes are not an important cofactor in the methyl group transfer reactions. yet understood. Attempts to model an association between Conclusions: Overall these results show that the metabolome stress and altered ethanol consumption in animals have not of human post-mortem frontal cortex samples is altered in in been successful. Our hypothesis is that individual diferences persons with history of excessive alcohol use when compared in stress efects on ethanol consumption are mediated by to controls. genetics. Methods. We measured alcohol consumption, using drin- Effects of a mindfulness based relapse prevention king-in-the-dark (DID) in females from two inbred mouse intervention on alcohol consumption and gut strains, C57BL/6J (B6) and DBA/2J (D2) and 35 of their microbial diversity: Preliminary findings inbred progenies (the BXD family). A control group was Hollis C. Karoly1, Sarah L. Hagerty1, Raeghan L. Mueller1, maintained under normal housing and a stress group was ex- Angela D. Bryan1, Kent E. Hutchison1 (1 Boulder, USA) posed to chronic mild stress (CMS), consisting of unpredic- table stressors over seven weeks. Alcohol intake was measured Emerging research suggests that mindfulness-based interven- over sixteen weeks in both groups during Baseline (preceding tions (MBIs) infuence the inhibitory control network, which 5-week period), CMS (intervening 7-week period), and post- is critical to the etiology and maintenance of alcohol use stress (fnal 4-week period). disorders (AUDs). Mindfulness is associated with alterations Results. Tere was a strong efect of CMS on alcohol intake. in the microbiota-gut-brain-axis (MGBA) across numerous A few strains demonstrated CMS-related increased alcohol patient populations (1-3), and gut and immune alterations consumption; however, most showed decreased intake. We especially impact brain regions involved in executive function identifed one suggestive quantitative trait locus on chromo- and inhibition (4, 5). We compared an 8-week Mindfulness some 5 that contains the neuronal nitric oxide synthase gene Based Relapse Prevention program to a Relapse Prevention (Nos1). Te expression of Nos1 is frequently changed fol- (RP) intervention on alcohol consumption and gut microbial

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 381 Congrès diversity in an AUD sample. 36 participants have com- AA participation and extent of social support system from pleted treatment so far. From baseline to post-treatment, the 1st survey as predictor variables, depression score from participants reduced monthly binge drinking (M=7.0 to 3.0; the Patient Health Questionnaire (PHQ-9) as the media- t(60)=2.25, p=.03), total drinks (M=112.2 to 74.7; t(62)=2.41, ting variable, and Alcohol Use Disorders Identifcation Test p=.02) and alcohol-related problems using the Alcohol Use (AUDIT-C) score as the dependent variable. Disorder Identifcation Test (AUDIT), (M=19.3 to 13.2; Results: Te degree of social support system establishment (t(69)=3.51,p<.001). No signifcant group diferences were from the 1st survey negatively correlated with the depression observed. Although sample sizes at 3-month follow-up severity in the 3rd survey. Moreover, the duration in AA are small, preliminary analyses show greater reductions for from the 1st survey and the degree of depression from the MBRP participants for total drinks (d=.56) and AUDIT 3rd survey correlated with the severity of alcohol problem (d=.30). Post-treatment, gut microbial diversity was negati- from the 4th survey. Te model’s goodness of ft (?2=12.927, vely associated with AUDIT (r=-.288, p=.021). Change in df=10, P=0.228, IFI=0.926, CFI=0.898, RMSEA=0.050 microbial diversity pre to post treatment was negatively asso- (90% CI: [0.0000-0.117])) satisfed the acceptance criteria ciated with alcohol consumption on the Alcohol Dependence proposed by Hu & Bentler (1999). Te regression coefcient Scale (r=-.246, p=.050), indicating that as consumption from this model show that the degree of depression from decreased, microbial diversity increased. While these preli- the 3rd survey is decreased as the degree of social support minary analyses are encouraging, the full sample is needed system establishment from the 1st survey increases (ß=-3.186, to determine whether MBRP is superior to RP and whether P<0.01). Increased severity of depression, resulting from efects are mediated in part by alterations within the MGBA. weak social support system, increased the severity of alcohol Future work characterizing the microbiota at each level of the problem (ß=0.152, P<0.01). Increases in the duration in AA phylogenetic tree will allow for characterization of species decreased severity of alcohol problem without the media- that are diferentially altered by alcohol use and MBIs. tion of depression (ß=-0.039, P<0.05). Among the control References variables, the alcohol problem severity from the 1st survey 1 - Househam AM, Peterson CT, Mills PJ, Chopra D. The Effects of showed positive auto-regression efect (ß=0.311, P<0.01). Stress and Meditation on the Immune System, Human Microbiota, and When the auto-regression efect by alcohol problem was Epigenetics. Adv Mind Body Med. 2017 ; 31 (4) : 10-25. http://www.ncbi. controlled, the degree of social support system establishment nlm.nih.gov/pubmed/29306937. Accessed March 27, 2019. st 2 - Kaliman P, Álvarez-López MJ, Cosín-Tomás M, Rosenkranz MA, Lutz A, from the 1 survey afected later alcohol problem through the Davidson RJ. Rapid changes in histone deacetylases and inflammatory gene mediation of depression. expression in expert meditators. Psychoneuroendocrinology. 2014 ; 40 : 96- Conclusion: Adequate social support system relieves depres- 107. doi:10.1016/J.PSYNEUEN.2013.11.004. 3 - Schnorr SL, Bachner HA. Focus: Microbiome: Integrative Therapies in sion and improvement in depression helps the recovery Anxiety Treatment with Special Emphasis on the Gut Microbiome. Yale J process of patients with alcohol use disorder. Longer partici- Biol Med. 2016 ; 89 (3) : 397. https://www.ncbi.nlm.nih.gov/pmc/articles/ pation in AA can have a persisting efect on alleviating alco- PMC5045149/. Accessed March 27, 2019. 4 - Alfonso-Loeches S, Pascual-Lucas M, Blanco AM, Sanchez-Vera I, Guerri hol problem. Terefore, combining support for establishing C. Pivotal role of TLR4 receptors in alcohol-induced neuroinflammation and sufcient social support system and psychosocial interven- brain damage. J Neurosci. 2010 ; 30 (24) : 8285-95. tions, such as AA, is important for the recovery of alcohol 5 - Guerri C, Pascual M. Mechanisms involved in the neurotoxic, cognitive, use disorder. Particularly, screening and providing treatment and neurobehavioral effects of alcohol consumption during adolescence. Alcohol. 2010 ; 44 (1) : 15-26. doi:10.1016/J.ALCOHOL.2009.10.003. for patients who are at high risk for depression are needed in order to achieve a successful recovery. References Depression mediating the effect of social support - Kelly JF, Hoeppner B, Stout RL, Pagano M. Determining the relative and Alcoholics Anonymous on alcohol use disorder importance of the mechanisms of behavior change within Alcoholics recovery in Korea: A 2-year longitudinal study Anonymous: a multiple mediator analysis. Addiction. 2012 ; 107 : 289-99. Seon Wan Ki1, Il Ho Park1, Won Mi Jung2, Jeong Seok3, - Moos RH, Moos BS. Protective resources and long-term recovery from alcohol use disorders. Drug and Alcohol Dependence. 2007 ; 86 : 46-54. 2 1 2 Soo Bi Lee ( Incheon Metropolitan City, Korea, Seoul, Korea, - Hong JW, Noh JH, Kim DJ. The prevalence of and factors associated with 3 Chung-ju, Korea) high-risk alcohol consumption in Korean adults: The 2009–2011 Korea National Health and Nutrition Examination Survey. PlosOne. 2017. https:// doi.org/10.1371/journal.pone.0175299. Purpose: In order to understand the factors contributing to the course of alcohol use disorder in South Korea, we Does a glucagon-like peptide 1 (GLP-1) conducted a nation-wide longitudinal follow-up study of receptoragonist reduce alcohol intake in patients alcohol use disorder in South Korea. Te mediating efect of with alcohol dependence? depression on factors infuencing the recovery of alcohol use Mette K. Klausen1, Mathias E. Jensen1, Marco Møller1, disorder was examined in this study. Anne-Marie Østergaard Grindsted Jensen1, Ninale Dous1, Methods: Biannual survey and clinical follow-up were Kamilla W. Miskowiak1, Patrick M. Fisher1, Gerda K. Thomsen1, conducted in patients with alcohol use disorder from the Sabine Vollstädt-Klein2, Ulrik Becker3, Claus Ekstrøm1, hospitals/clinics and community mental health centers Gitte M. Knudsen1, Tina Vilsbøll4, Anders Fink-Jensen1 representing 6 districts in South Korea between 2016 and 1 2 2017. Data of 120 individuals who complete all four surveys ( Copenhagen, Denmark, Mannheim/Heidelberg, Germany, 3 4 were analyzed. Path analysis was conducted with duration of Hvidovre, Denmark, Gentofte, Denmark)

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Aims: Glucagon-like peptide-1 (GLP-1) receptorstimulation and cell/neuronal loss in midline thalamus in adulthood, after has proven to reduce alcohol consumption in preclinical ex- the rats underwent comprehensive behavioral testing. periments with rodents and non-human primates. However, Our data indicate that AE during third trimester equivalent the efect of GLP-1 receptor agonists on alcohol reduction leads to 30-fold increase in cell death and cell loss (≈30%) in in humans with alcohol dependence has to ourknowledge,not Re but not in neighboring thalamic nuclei (mediodorsal and yet been investigated. rhomboid nuclei). Tis cell loss is driven by loss of neurons Methods: Te efect of the once-weekly GLP-1-receptor- that persists into adulthood. AE rats display alterations in agonist, will be investigated in adouble-blinded, object-in-place memory and impairments in rule switching placebo-controlled, randomized clinical trial. 114 outpatients, in a plus maze-based operant conditioning task in adulthood. age 18-70 years will be randomized to eitherplacebo orexe- Tese data suggest that Re is specifcally targeted by postnatal natide once-weekly for 26 weeks as a supplement to cogni- AE and that this damage persists into adulthood. Te inte- tivebehaviouraltherapy. Te primary endpoint is reduction grity of RE may be a structural indicator of impaired execu- in numberof ‘heavy drinking days’, measured by the Time tive functioning observed in some manifestations of FASD. Line Follow Back (TFLB) method. Secondary endpoints include changes in total alcohol consumption, days without Tuftsin peptide analog prevents ethanol-induced consumption, changes in brain activity and function, smoking cognitive impairment in aged alcohol-withdrawn status, cognition, measures of quality of life and changes rats through modulation BDNF signaling pathway in phosphatidylethanol (PEth) as a biomarkerof alcohol L.G. Kolik1, A.V. Nadorova1, T.A. Antipova1, S.V. Kruglov1, consumption from baselineto follow-up at week 26. V.S. Kudrin1, A.D. Durnev1 (1 Moscow, Russia) In addition to these clinical outcomeparameters, we willex- plore the possible neurobiological underpinnings by use Background. (Thr-Lys-Pro-Arg-Pro-Gly-Pro), as of functional Magnetic Resonance Imaging (fMRI) and tuftsin analogue, is safe anxiolytic with cognitive enhancing the possible neuromolecular changes in striatal dopamine properties. However, there is no data about its use in patients transporter(DAT) availability by use of the Single photon with comorbid alcohol use disorders, so, work aimed to study emission computed tomography (SPECT). selank efects in early alcohol withdrawal. Results: 103/114 patients are recruited. Methods. Male albino rats were administered 10% (v/v) Conclusions: Te potential as a new treatment is being tested. ethanol (EtOH) as the only source of drinking water within If successful, this could be anew revolutionary treatment 30 weeks (n=20). Ten EtOH-withdrawn rats were treated foralcohol dependence. saline (“EtOH”) and selank 0.3 mg/kg, i.p. (“EtOH+selank”) Financial support: The study is financed by Region for 7 days. EtOH-naïve age-matched rats (n=20) were treated Hovedstadens Forskningsfond, Region Hovedstadens by saline (“Control”) and selank (“Selank”). Learning capaci- Psykiatri and Fonden Novavì. Te manufacturerof Bydureon®, ties were measured in novel object recognition task 24 hours AstraZeneca A/S, has no fnancial interest or involvement in after selank treatment, then rats were sacrifced. Selected this project. brain neurochemicals were measured by HPLC, expression of BDNF protein in particular brain structures was analyzed Why me? One midline thalamic nucleus is using Western blot. critically important for hippocampo-prefrontal Results. Selank prevented decrease of discrimination index cortex communication and vulnerable to alcohol of novel object (p<0.05) in aged EtOH-naïve and EtOH- exposure during third trimester equivalent withdrawn rats indicating its positive impact on cognitive Anna Klintsova1 (1 Newark, USA) performance. Selank restored increased 5-OT and reduced turnover 5-HIAA/5-OT in frontal cortex, prevented alco- Children exposed to alcohol in utero display physical and hol-induced increased aspartic acid, glycine and taurine levels behavioral irregularities, including impairment in “executive in hypothalamus, GABA in n.Acc. and aspartic acid and function” (EF) behaviors that require coordination between glycine in striatum. Forced alcohol intake with subsequent prefrontal cortex (PFC) and hippocampus (HPC). Non- withdrawal led to signifcant increase in BDNF level in human primate and rodent studies have demonstrated that hippocampus (“Control” 2.0±0.2 R.D.U., “EtOH” 3.4±0.6 the midline thalamic nucleus reuniens (Re) is essential in R.D.U.) and frontal cortex (“Control” 1.1±0.2 R.D.U., coordinating PFC-HPC activity, as selective Re inactivation “EtOH” 1.8±0.3 R.D.U.). Selank prevented BDNF increase impairs PFC-HPC synchrony and behavioral performance. in hippocampus and frontal cortex. To unveil the structural defcits in HPC-Re-mPFC circuitry, Conclusions. The obtained results indicate pronounced one needs to consider that Re is a critical intermediary with efect of taftsin analog on age-related changes associated reciprocal connections to mPFC and HPC, while mPFC with memory impairment, accompanied by chronic alcohol connects with HPC via Re. intoxication possibly through modulation BDNF signaling Rodent model of binge drinking during third trimester was pathway. used in our studies: rat pups were intubated with moderate or Alcohol-related mortality in the WHO European high doses of alcohol (AE) or sham-intubated (SI) on PD4-9. Unbiased stereology was used to estimate cell death after AE region: Sex-specific trends and predictions

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1 1 1 Daniel König , Nathalie Pruckner , Andrea Gmeiner , Barbara intervention and passive informing indicates a signifcant 1 1 1 Hinterbuchinger , Matthäus Fellinger , Thomas Waldhör , decrease in the frequency of alcohol consumption. At 3 1 1 1 Otto M. Lesch , Stephan Listabarth , Andreas Wippel , Sandra months follow-up, signifcant diferences were found between 2 1 1 2 Vyssoki , Benjamin Vyssoki ( Vienna, Austria, St. Pölten, Austria) the experimental and control groups: 47% of the women in the experimental sample and 62% in the control group were Introduction. According to data from the WHO Health at risk. After 6 months, the diferences are found at the level For All Database, alcohol-related deaths have signifcantly of the statistical tendency (45% and 55%, respectively), and decreased within the European region between 1979 and after 12 months no signifcant diferences were revealed (46% 2015. Yet, there are still pronounced diferences between and 49%, respectively), which indicates a faster efect achie- regions and burden of alcohol consumption and dependence ved with the brief intervention method. remains high. Tus, the results of the study indicate the efectiveness of the Aims. Alcohol is an important risk factor for morbidity and brief intervention designed to prevent FAS and FASD. mortality, especially within the European region. Diferences in per capita consumption and drinking patterns are possible What does impact health related quality of life in reasons for regional diferences and diverging trends in alco- alcohol use disorder: Cognitive deficits, anxiety or hol-related health outcomes. depression? Methods. For 29 countries within the WHO European Najlaa Lahbairi1 , Alice Laniepce1, Nicolas Cabé1, region the last four decades were evaluated for trends and François Vabret1, Céline Boudehent1, Géraldine Rauchs1, predictions in alcohol-related deaths using data available Anne-Lise Pitel1 (1 Caen, France) from the WHO Health For All Database. Results. Between 1979 and 2015, age-standardised death Alcohol Use Disorder (AUD) results in multiple social and rates for both sexes due to selected alcohol-related causes cognitive problems with a poor health related quality of life decreased signifcantly in all included countries of the WHO (HRQoL). Te association between HRQoL and cogni- European region, but regional diferences were still pro- tion is well-known in various diseases (stroke, dementia...). nounced. Assuming a similar trend in the future, the model While HRQoL is crucial to maintain abstinence, it remains predicted a further decrease until the year 2030. little studied in AUD. Depression and anxiety also afect Conclusions. Even though alcohol-related mortality might HRQoL and cognition, potentially exacerbating the risk may have decreased within the last decades, the detrimen- of relapse. Te objective of this study was to investigate tal efects of alcohol consumption and alcohol dependence the relationships between HRQoL, cognition and mood remain a considerable burden of disease within Europe. in AUD. Tirty-three recently detoxifed AUD inpatients Tis study provides information on possible reasons why and 28 healthy control (HC) subjects were included. An some countries show greater and others show lower decreases extensive neuropsychological assessment was conducted and on alcohol-related mortality. To put light on these various the intensity of depressive and anxiety symptoms was mea- – and especially – infuenceable factors, further research is sured using the Beck Depression Inventory (BDI) and the recommended. Findings on this area, such as certain legal Spielberg State-Trait Anxiety Inventory (STAI). In AUD regulations or adequate interventions to reach potentially patients, HRQoL was evaluated using the Alcohol Quality of alcohol burdened people earlier, are from utmost importance Life Scale (AQoLS), which focuses on 7 diferent domains: to establish essential preventive strategies. activities, relationships, living conditions, negative emotions, Brief intervention aimed at fetal alcohol syndrome looking after self, control and sleep. Compared to controls, prevention: Efficacy study AUD patients showed higher levels of depression, anxiety Ekaterina A. Burina1, Almara K. Kulieva1 (1Saint Petersburg, Russia) and more severe cognitive impairments. All patients complai- ned on at least 6 of the 7 domains of HRQoL. Contrary to our expectations, HRQoL did not relate to cognition, but to Tis study focuses on the psychological efects of brief inter- depression (r=0.53, p=0.02) and anxiety (r=0.42, p=0.01). Our ventions aimed at preventing Fetal Alcohol Syndrome (FAS) results confrm previous fndings suggesting altered HRQoL, and Fetal Alcohol Spectrum Disorders (FASD). impaired cognitive abilities and altered mood in AUD Te sample of the study consisted of 280 women of child- patients early in abstinence. Te pattern of a relationships bearing age: 140 women entered the experimental group and between HRQoL, cognition and mood suggests that impro- 140 – the control group. All participants were screened; a ving HRQoL in AUD patients may require prioritizing the basic interview and three follow-up interviews at 3, 6 and treatment of anxiety and depression. 12 months were conducted. All women received information materials (a brochure) about the alcohol efects on the fetus Withdrawal history is associated with sleep and and fetal alcohol syndrome. With women of the experimental cognitive alterations in recently detoxified alcohol group, after a baseline interview, twice in the period from 2 use disorder patients weeks to one and a half months, specially trained OBGYN Alice Laniepce1, Nicolas Cabé1, Françoise Bertran1, Claire André1, physicians carried out a dual-focused brief intervention. 1 1 1 Te dynamics of the actual alcohol consumption by women Céline Boudehent , François Vabret , Francis Eustache , 1 1 1 of childbearing age under the infuence of dual-focused brief Géraldine Rauchs , Anne-Lise Pitel ( Caen, France)

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Background: Early in abstinence, patients with Alcohol Use participants show diferent subjective and psychophysiologi- Disorder (AUD) frequently present brain alterations, cogni- cal responses to appetitive cues during an alcohol cue reacti- tive defcits and sleep disturbances (1, 2). Considering the vity task compared to placebo participants, and whether these crucial role of sleep in cognitive functioning, we aimed at responses are associated with prospective drinking outcomes. investigating whether objective sleep disturbances contribute Method: Forty-two alcohol dependent participants (pla- to cognitive defcits in AUD patients recently detoxifed. cebo: n=12, low-dose baclofen [30 mg/day] n=18, high-dose Given the short delay since drinking cessation, relationships baclofen [75 mg/day]: n=12) completed an alcohol cue with withdrawal history were also explored. reactivity task, whereby water and alcohol beverage cues Methods: 18 AUD patients underwent a neuropsychological were presented, with subsequent respective recovery periods. battery and sleep examinations (including a 1-week conti- Subjective alcohol craving and psychophysiological indices nuous actigraphy recording and one night of polysomno- (skin conductance; cardiovascular measures: heart rate, high- graphy). Withdrawal history was also documented. While frequency heart rate variability) were recorded across the task. patients were early in abstinence, none of them presented Results: High-dose baclofen-treated participants showed physical symptoms of alcohol withdrawal (3) nor were under both overall cue reactivity to both water and alcohol cues and medication by . Regressions analyses were greater recovery efects during recovery periods, revealed by performed between cognition, sleep, and withdrawal history. high-frequency heart rate variability levels, when compared Results: Longer sleep duration was associated with a lower to low-dose- and placebo-treated participants. Tere were no amount of slow-wave sleep and executive defcits in recently medication efects on subjective alcohol craving. In high-dose detoxifed AUD patients. Alcohol withdrawal history, espe- baclofen participants only, there was a predictive efect of cially the duration of benzodiazepines prescription and the lower baseline resting heart rate variability and fewer post- total amount of benzodiazepines prescribed, was related to test percentage of heavy drinking days. sleep abnormalities and executive defcits. Discussions and conclusions: Tere was a dose-specifc res- Discussion: Our results suggest that alcohol withdrawal and cuing efect of high-dose baclofen on the dynamic modula- associated benzodiazepines prescription result in poor resto- tion of reactivity and regulation of responses to eliciting cues. rative sleep and executive defcits in recently detoxifed AUD Using psychophysiological techniques to detect treatment patients. Te duration and severity of alcohol withdrawal responses may elucidate baclofen’s mechanisms of action, and should be taken into account when a neuropsychological potentially identify alcohol use disorder subgroups that may assessment is conducted early in abstinence. Further studies best beneft from this pharmacotherapy. are required to disentangle the neurotoxic efect of with- Disclosure of interest statement: Te Australasian Professional drawal per se from the consequences of the benzodiazepines Society for Alcohol and other Drugs (APSAD) recognises treatment. the considerable contribution that industry partners make References to professional and research activities. We also recognise the 1 - Le Berre AP, Laniepce A, Segobin S, Pitel AL, Sullivan EV. Alcohol Use Disorder: Permanent and Transient Effects on the Brain and need for transparency of disclosure of potential conficts of Neuropsychological Functions. The Oxford Handbook of Adult Cognitive interest by acknowledging these relationships in all written Disorders. 2019. 302-32. publications. Tere are no competing interests related to this 2 - Chakravorty S, Chaudhary NS, Brower KJ. Alcohol Dependence and Its study. Relationship With Insomnia and Other Sleep Disorders. Alcohol Clin Exp Res. 2016 ; 40 : 2271-82. 3 - Cushman PJ, Forbes R, Lerner W, Stewart M. Alcohol withdrawal syn- Affect preceding drinking sessions predicts dromes: clinical management with lofexidine. Alcohol Clin Exp Res. 1985 ; increased alcohol consumption in University 9 : 103-8. students: an experience sampling approach. Warren Logge1, Benjamin Riordan1, Kirsten Morley1, What can we learn from epidemiology in alcohol Andrew Baillie1, Paul Haber1, Tamlin Conner2 (1 Sydney, Australia, research? Recent findings from large population- 2 Dunedin, New Zealand) based cohorts Cédric Lemogne1 (1 Paris, France) Introduction: University students are a high-risk group for developing alcohol problems. Positive and/or negative afect Baclofen modulates psychophysiological responses is associated with increased consumption, but there are mixed to appetitive cues in treatment-seeking alcohol results. use disorder individuals Impulsivity, which is a key risk factor for initiation of and Warren Logge1, Andrew Baillie1, Paul Haber1, Kirsten Morley1 excessive alcohol use, may explain the link between afect (1 Sydney, Australia) and drinking. Tis study used experience sampling to assess whether reported afect prior to drinking was associated with Introduction: Baclofen is an emerging potential pharmaco- increased consumption, and whether impulsivity moderated therapy for alcohol use disorder. Little research has investiga- this association. ted how baclofen afects psychophysiological responses to al- Method: We recruited 694 University students (18-25 years) cohol cues, and subsequent efects upon drinking behaviours. for a micro-longitudinal daily diary study, with impulsivity We assessed whether baclofen-treated alcohol dependent (BIS/BAS) measured at baseline. Students reported afect

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(positive, negative) via text message four times per day for 13 tamate following baclofen treatment in alcohol dependent days, and daily alcohol use. individuals. Results: Linear mixed models found a three-way interaction Disclosure of interest statement: Tere are no competing between positive afect, number of drinking days, and the interests related to this study. BIS/BAS Drive subscale score. For participants who drank less frequently, those with higher Drive scores reported a The active ingredients of Bupleurum falcatum, higher number of drinks per session with increasing positive saikosaponins A and D, but not C, reduce alcohol afect, while those lower Drive scores showed less pronounced self-administration in rats increase. For participants who drank more frequently, those Paola Maccioni1, Federica Fara1, Irene Lorrai1, Young-Won Chin2, with higher Drive scores showed little change regardless of Jung Hwan Lee3, Hak Cheol Kwon3, Giancarlo Colombo1 positive afect, whereas those with lower Drive scores showed (1 Monserrato, Italy, 2Goyang, Gyeonggi-do, Republic of Korea, a marked increase in drinks per session according to positive 3 Gangneung-si, Gangwon-do, Republic of Korea) afect. Tere were no efects found related to negative afect. Discussions and conclusions: Positive afect, but not negative Treatment with saikosaponin A (SSA) – an active ingre- afect, has a key role in consumption levels according to dient of the medicinal herb, Bupleurum falcatum – has been drinking session frequency and level of goal-directed moti- reported to suppress i.v. self-administration of and vation in university students. Tis association is complex and cocaine and oral self-administration of alcohol in rats. It has dependent on drive and established patterns of drinking. been demonstrated that these anti-addictive properties of Disclosure of interest statement: Tere are no competing SSA occur, at least in part, via a GABAB receptor-mediated interests related to this study. mechanism. Tis lab has recently started a research program aimed at investigating whether ingredients of Bupleurum Brain metabolites and hypothalamic-pituitary- falcatum other than SSA afect alcohol self-administration adrenocortical activity during baclofen treatment in rats. Accordingly, the present study investigated whether in alcohol dependent patients: Modulation by the the anti-alcohol properties of SSA extend to saikosaponin GABAB receptor polymorphism rs29220 C (SSC) and saikosaponin D (SSD; an epimer of SSA). To Kirsten C. Morley1, Natasha Luquin2, Jim Lagopoulos1, this end, adult female Sardinian alcohol-preferring (sP) rats Warren Logge1, Andrew Baillie1, Ronald J. Trent2, Paul S. Haber1, 2 were trained to lever-respond for alcohol (15%, v/v) on a fxed (1 Sydney, Australia, 2 Camperdown, Australia) ratio 5 (FR5) schedule of reinforcement. Once responding had stabilized, rats were tested under the same schedule after Introduction: We have previously shown that the GABBR1 treatment with saikosaponins. Treatment with SSA (0.25-1 rs29220 polymorphism is associated with response to baclo- mg/kg, i.p.) and SSD (0.25-1 mg/kg, i.p.) resulted in highly fen, a GABAb agonist, in the treatment of alcohol de- similar, marked reductions (50-60% at the highest dose pendence. In the current study, we aimed to further examine tested) in lever-responding for alcohol and amount of self- the role of the GABBR1 rs29220 polymorphism on hypotha- administered alcohol. Conversely, treatment with SSC (0.25- lamic-pituitary-adrenocortical activity and neurometabolites 1 mg/kg, i.p.) failed to alter lever-responding for alcohol and following administration of baclofen (BAC) or placebo (PL) amount of self-administered alcohol. Future experiments will in alcohol dependent individuals. investigate the efect of other saikosaponins on alcohol self- Method: Parietal GABA, Glutamate, Glutathione and administration in sP rats, with the intent of establishing a N-Acetyl Apartate levels were measured in N=25 alcohol possible structure-activity relationship. Tese results confrm dependent patients using in vivo proton magnetic resonance that Bupleurum falcatum is a valuable source of compounds spectroscopy (1H-MRS) 120 minutes following adminis- with anti-alcohol potential. tration of PL or BAC. Blood samples were obtained for analysis of the single nucleotide polymorphism (rs29220) in Is it relevant to postpone psychosocial treatment the GABAB receptor subunit 1 gene (GABBR1) (CC=15, of alcohol dependence by one month to favor G-=10). Plasma cortisol levels were also measured at two early neuropsychological recovery? time points including pre and post scan. Angéline Maillard1, Hélène Poussier2, Céline Boudehent1, Results: Tere was a signifcant efect of medication (BAC vs Coralie Lannuzel1, Angel Vicente2, François Vabret1, PL) on cortisol levels (F=10.18, p=0.007) but there were no Nicolas Cabé1, Anne-Lise Pitel1 (1 Caen, France, 2 Ifs, France) signifcant main efects of genotype (G- x CC) or medica- tion (BAC vs PL) x genotype (G- x CC) interaction efect. Many recently detoxified Alcohol Use Disorder (AUD) Tere was a signifcant medication (BAC vs PL) x genotype patients early in abstinence exhibit neuropsychological im- (G- x CC) interaction efect for parietal concentrations of pairments which limit the beneft of treatment and increase glutamate (F=4.87, p=0.04) but not for the other metabolites. the risk of relapse. While psychosocial alcohol treatment may Discussions and Conclusions: Our data demonstrate that the not be clinically relevant in AUD patients with impaired GABBR1 rs29220 polymorphism does not moderate baclofen neuropsychological abilities, it is now clear that these neuro- induced changes in HPA axis activity. Te GABBR1 rs29220 psychological defcits can be partially or totally reversible polymorphism did moderate cortical concentrations of glu- with drinking cessation. Te main purpose of this retrospec-

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 386 Congrès tive clinical study was to investigate whether a three-week conclusion, a promising peripherally selective CB1 receptor stay as inpatients in a convalescent home enables neuro- antagonist has been identifed that is suitable for further cli- psychological defcits observed in recently detoxifed AUD nical development for ALD and other disorders. patients to recover and even to return to normal. Neuropsychological data were collected in 84 AUD patients. Telomere length and polymorphisms in telomerase Five neuropsychological components were assessed before genes among patients with alcohol use disorders and after a three-week multidisciplinary treatment in conva- C. Carbonell1, H. Llorente1, E. Bueno1, J. A. Pérez Rivera2, lescent home. Baseline and follow-up performance was com- M. A. Pérez1, C. Cieza1, J. Fernández Mateos1, pared using Wilcoxon’s and Chi-square tests. R. González-Sarmiento1, F. Laso1, M. Marcos1 (1 Salamanca, The comparisons between baseline and follow-up per- Spain, 2 Burgos, Spain) formance revealed a signifcant improvement for the fve cognitive components. Te ratio of patients with preserved Introduction: Telomeres are repetitive DNA sequences loca- or impaired performance was signifcantly diferent between ted at the ends of chromosomes, protecting cells from loss of the baseline and follow-up sessions for three components, genomic material during replication. Telomere length (TL) indicating that there were fewer patients with impaired per- variation is associated with many infammatory diseases. formance at follow-up than at baseline. Te relationship between alcohol consumption and TL has In recently detoxifed AUD patients, impaired cognitive func- been previously studied, but, to date, no clear relationship is tions recover with a three-week stay in a convalescent home established regarding alcohol use disorders (AUDs) and TL. ensuring sobriety and healthy nutrition. Such therapy seems Polymorphisms in telomerase genes have also been associa- favoring cognitive recovery and even performance to return ted with susceptibility to AUDs. Here, we have analysed TL to normal. It is thus crucial to postpone alcohol treatment for and the polymorphisms TERC rs2293607, rs12696304, and AUD patients with neuropsychological impairments in order rs16847897, TERT rs2735940, rs2736100, and rs2736098 in to make them cognitively able to beneft from it. patients with AUDs. Development of a peripherally restricted CB1 Patients and methods: 99 men with AUDs (according to DSM IV criteria) and 99 healthy age and sex-matched receptor antagonist for alcohol induced liver controls were included. DNA was extracted from peripheral disease blood leukocytes using phenol/chloroform procedure and George Amato1, Amruta Manke1, Robert Wiethe1, Vineetha 1 1 1 1 genotyped using TaqMan 5’-exonuclease allelic discrimi- Vasukuttan , Rodney Snyder , Yun Lan Yueh , Ann Decker , nation assays (Applied Biosystems). Relative mean TL was 1 1 1 1 Scott Runyon , Nayaab Khan , Rangan Maitra ( Durham, USA) measured from DNA by a qPCR assay. Statistical analysis was performed using GenEx v6 software and SPSS v25. Antagonists of peripheral type 1 cannabinoid receptors (CB1) Results: Mean telomere length (T/S) was 4.38±3.18 in alco- can treat various diseases including alcoholic liver disease holic patients and 6.28±1.94 in controls (p<0.001). Alcoholic (ALD). Unfortunately, inhibition of human CB1 (hCB1) patients with alcoholic liver cirrhosis or without liver disease receptors in the central nervous system (CNS) produces had also shorter TL than their respective controls. Area adverse efects including depression, anxiety and suicidal under ROC curve to study correlation between telomere ideation. Terefore, eforts are underway to develop periphe- length and alcoholism was 0.70 (p<0.001) and the best cut- rally restricted antagonists of hCB1. Recent crystal structures of-point of telomere length (T/S) was 5.91 (sensitivity 70% of hCB1 and docking studies with the purine otenabant, a and specifcity 64%). Te presence of the G allele of allelic centrally acting CB1 inverse agonist developed by Pfzer that variant TERC rs2293607 polymorphism was associated with was clinically tested but abandoned due to concerns related increased risk of AUDs and shorter TL. to adverse efects, indicated that the piperidine group of this Conclusion: Our study supports the correlation between TL compound could be functionalized at the 4-position to access attrition and chronic and excessive ethanol intake, which has a binding pocket that might accommodate both polar and not been previously analyzed in this population. In addition, nonpolar groups. Terefore, we proceeded to examine the the possession of the G allele of TERC rs2293607 allelic piperidine as a linker, which was functionalized with alkyl, variant is associated with both shorter TL and increased heteroalkyl, aryl and heteroaryl groups using a urea connector. risk of AUDs and therefore may be associated with genetic Tese studies resulted in orally bioavailable and peripherally susceptibility to develop alcoholism through modulating TL. selective compounds that were potent inverse agonists of hCB1 with exceptional selectivity for hCB1 over hCB2. Funded by ISCIII and FEDER PI16/01548 and Te lead compound from this series presented good ADME RD16/0017/0023 properties, clean selectivity profle against >40 high-risk Impact of chronic alcohol exposure and receptor targets (SafetyScreen, Eurofns), and was advanced withdrawal on hepatocellular carcinoma into in vivo efcacy studies in the Lieber DeCarli model of aggressiveness alcohol-induced steatosis. Once a day oral dosing with this Constance Marié1, Charles-Antoine Papillon1, Grégory Fouquet1, lead compound blocked alcohol-induced liver steatosis in 1 1 1 mice and reduced expression of several molecular biomarkers Hicham Bouhlal , Éric Nguyen-Khac , Mickael Naassila , 1 1 associated with hepatic infammation and metabolism. In Ingrid Marcq ( Amiens, France)

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In France, 9.7% of the population drinks alcohol every day. Effect of behavioral “super-intervention” in Futhermore, 50% of liver disease mortality is due to alcohol. adolescence on cortically-projecting cholinergic Indeed, chronic alcohol exposure can lead to HepatoCellular neurons in a rodent model of Fetal Alcohol Carcinoma (HCC) development. HCC is the 6th cancer in Spectrum Disorders (FASD) incidence but the 2nd cancer in mortality. HCC patients have K.A. Milbocker1, Z.H. Gursky1, A.Y. Klintsova1 (1 Newark, USA) a 5-year survival lower than 10 percent. Te study of C.E. Costentin (et al, 2018, Cancer) showed a One in twenty infants in the United States are afected decrease of survival in patients with alcoholic HCC. Tese by prenatal alcohol exposure (PAE) resulting in a range of observations determined that alcoholic HCC are more disorders categorized as Fetal Alcohol Spectrum Disorders aggressive than other etiologies of HCC. Tis study also (FASD). PAE during late-stage pregnancy can produce las- demonstrates the importance of withdrawal in patient sur- ting defcits of cortical-dependent behaviors in afected indi- vival. Our project aims to determine the physiopathological viduals (executive function, i.e. decision-making and inhibi- mechanisms explaining these clinical observations. tory control). Evidence from human cases and animal models We developed a protocol of Chronic Alcohol Exposure of FASD suggest that cognitive impairments are mitigated by (CAE). For this, HCC cells were exposed to alcohol during non-invasive intervention such as choline supplementation several months at diferent doses. We also devised a with- (Fuglestad et al., 2013; Idrus et al., 2017). drawal method simultaneously. We investigated the impact Our study examines the efects of a behavioral “super-inter- of alcohol on cellular viability, mortality and proliferation. vention” on the integrity of cortically-projecting cholinergic Furthermore, we studied cancer stem cell markers in cells neurons from the nucleus basalis of Meynert (NBM) in a exposed to CAE and after withdrawal, and fnally their rat model of FASD. Male Long-Evans rats were exposed to efects on migratory and invasive cell potentials. binge-like alcohol (5.25 g/kg/day) on postnatal days (PD) Our results show that CAE decreases cellular viability and 4-9. Control groups included sham-intubated (SI, no liquid) promotes aggressiveness by an increase of metastatic potential and suckle-control (SC) rats. Rats were weaned on PD23. and cancer stem cell markers. Interestingly, withdrawal par- On PD30, behavioral “super-intervention” began, consis- tially reverses these modifcations due to CAE. ting of twelve days of voluntary running in a wheel (WR) followed by four weeks of housing in a complex environ- Our results obtained in HCC cellular model exposed to CAE ment (EC). Immunocytochemical visualization of ChAT+ allow a better understanding of the mechanisms underlying neurons was performed on brain tissue collected at PD72. decreased survival of patients with alcoholic HCC. Tey also Preliminary analyses of unbiased stereological estimates demonstrate the importance of looking for alcohol abstinence show a signifcant increase in the number of cholinergic in patients. neurons following adolescent intervention (F(1, 27)=6.63, The Effects of environmental enrichment on p=0.02; interaction with treatment p=0.08) and volume of intermittent 20% ethanol intake in Long-Evans NBM (F(1, 27)=4.65, p=0.04). However, it appears that the Rats intervention afects postnatal treatment groups diferently 1 2 2 1 (SC: t(8)=2.86, p=0.02), indicating decreased neuroplasticity L. Shehata , J. Jeanblanc , M. Naassila , M.P. Martinetti in AE rats long-term. (1 Ewing, USA, 2 Amiens, France) Violations of a healthy behavior of pregnant Environmental enrichment (EE), such as toys or novel ob- women: Pilot study jects, is recommended practice for laboratory animal research; Valentina A. Moshkivskaya1, Ekaterina A. Burina1 (1Saint however, in animal models of alcohol consumption, EE has Petersburg, Russia) produced varying efects on ethanol (EtOH) intake. Te current study examined the efects of EE on EtOH intake Most women know that proper nutrition, avoiding the use in male Long-Evans rats using an intermittent two-bottle of various psychoactive substances, moderate exercise, pro- choice (2BC) paradigm. Animals were randomly assigned to longed sleep during pregnancy is a necessary condition for either an EE group (n=12), with a packet of crinkle paper the normal development of the fetus, the course of pregnancy in the home cage, or non-enriched control (n=12). A 20% and the health of mother and child. However, a number of EtOH solution was provided concurrently with water in the studies reveal a violation of the principles of healthy behavior. home cage for 24-hr periods on Mon, Wed, and Fri each In this regard, it was decided to conduct a study that allows week for 13 weeks. Overall, EE rats consumed less absolute investigating some of the habits of a violation of the healthy EtOH (g/kg/24 hrs) compared with CTRL rats. Tis efect lifestyle of pregnant women. was specifc to EtOH, as water consumption on the days Materials and research methods. 25 pregnant women took between the two-bottle choice periods did not difer, nor did part in the study: 20 – had a physiological pregnancy, 5 – got the groups difer with respect to body weight. Tese fndings pregnant with the help of reproductive methods. Te sample suggest that EE reduces 20% EtOH consumption in an was enrolled on the base of a maternity hospital. Social intermittent 2BC paradigm, and these results underscore the interview, analysis of medical notes and Questionnaire of experimental implications of environmental enrichment in Disorders of Healthy Behavior (Lutsenko, Gabelkova) was animal models of alcohol use disorder. used in the study due to research aim. Te questionnaire

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 388 Congrès includes 9 scales: smoking, eating disorders, neglecting safety, variable, easy and quick to asses and that can ofer more alcohol use, chasing a trendy image, low self-control, emo- information that we expected to. At this study we do not only tional incompetence, self-destructive behavior and general confrm that the amount of alcohol consumption that a student performance. who practices alcohol asses of him/herself highly corresponds Study results indicated low and average level of violation in with his/her real consumption, but we observed how this general. Detailed analysis showed a tendency of women with self-perception of alcohol use at 18 years old is an important frst pregnancy to have normal eating and drinking habits in predictor of the RAC and BD at 20, 22, 24 and 28 years old. comparison to women who already have one or two children Relationship between liver fibrosis and liver iron (W=113.5, p-value=0.0549). Participants with higher level of emotional incompetence tend to have more eating problems in patients with alcoholic liver disease 1 1 1 1 1 or disorders (r(25)=0.63, p=0.002). Current study should be J. Mueller , H. Raisi , O. Elshaarawy , V. Rausch , I. Silva , 1 1 1 continued to gain more crucial information that can be used H.K. Seitz , S. Mueller ( Heidelberg, Germany) for preventive measures. Background: Alcoholic liver disease (ALD) is one of the most Validity of the selfperception of alcohol common liver diseases and can cause hepatic iron overload. consumption among freshmen Aim: To investigate the relationship between fbrosis stage Lucia Moure-Rodriguez1 (1 Ourense, Spain) and liver iron in ALD patients. Methods: 358 patients were prospectively recruited between 2007 and 2018 at the department of Internal Medicine Aim: Our aim is to assess the importance of the perception at Salem Medical Center, Heidelberg. In 224 patients of university students about their own alcohol consumption with ALD, non-invasive liver stiffness (LS) (FibroScan, at 18 years of age on risky alcohol consumption (RAC) and Echosense, Paris) and iron determination (room temperature binge drinking (BD) at that same age and during the fol- susceptometry, RTS) was performed. 134 patients with liver lowing 10 years. biopsy had histological assessment of liver fbrosis (Kleiner) Methods: a cohort study among university students and iron (Prussian Blue stain). Additionally, all patients had (Compostela Cohort 2005, Spain), was carried out between routine laboratory tests and abdominal ultrasound. November 2005 and February 2015. Participants were selec- Results: Mean age was 52.4 years and mean alcohol consump- ted by cluster sampling, going to at least one 1st year class of tion was 176 g/day. Biopsied patients showed more fbrosis every faculty of the Universidade de Santiago de Compostela. than non-invasively characterized patients (fbrosis stage 1.7 Te Alcohol Use of Dissorder Identifcation Test was used to 2.6, P<0.001). No signifcant correlation between fbrosis/ for measuring both RAC, – total score ≥5 for women and LS and histological iron or RTS was observed, while ferritin ≥6 for men –, and BD – drinking ≥6 alcoholic beverages in was moderately associated with LS (r=0.45, P<0.01). Serum one single occasion at least monthly–. Students answered transferrin decreased continuously with LS/fbrosis (r=-0.51, questions related to their expectations about alcohol (Low/ P<0.001). Te relationship between fbrosis stage and liver High) and their perceptions of their own alcohol consump- iron was non-trivial in both cohorts. Liver iron (histology tion (nothing/little/fair amount/large amount). A multilevel and RTS) was elevated in both cohorts in fbrosis stages logistic regression was performed and predictive values were up to stage 3 while it was signifcantly decreased in fbrosis calculated with SPSSv20 statistical software. stage 4 in comparison to stage 3 (P<0.05). Tis decrease was Results: 99% of students in class the day of the survey partici- also observed in ferritin values, however, non-signifcantly. pated (n=1,382). As subject’s perception of their own alcohol Accordingly, RTS was signifcantly associated with LS in consumption at 18 years old increases, so does the proportion patients with stifness less than 12 kPa (r=0.22, P<0.05), of them with positive expectations regarding alcohol (55.2% while no signifcant correlation was observed in patients with vs 14.3%) and the proportion of them who practice RCA high LS >20 kPa. (100% vs 14% among females and 100% vs 37.7% for men) Conclusions: In contrast to common perception, ALD and BD (84.4% vs 10% among females and 85.7% vs12.3% patients with liver cirrhosis showed signifcantly lower liver on men) at this same age. Tis tendency is maintained in iron in histology and RTS. Te highest iron concentrations relation to their RCA and BD during 20, 22, 24 and 28 years were found in intermediate fbrosis stages F1 to F3. Elevated old among both genders. Taking in to account subjects who ferritin or lower transferrin in liver cirrhosis patients is more perceive their consumption was nothing or little at 18 years likely due to a release of iron from cell death instead of old compared to those who perceive that they consume a fair increased body iron stores. amount or large amount of alcohol, this last group maintains much higher prevalence of RCA and BD all through the Identification of novel epigenetic biomarkers of study period. Understanding one’s alcohol consumption per- alcohol dependence in brain and blood ception as a diagnostic tool, they were observed high negative Vanessa Nieratschker1 (1 Tuebingen, Germany) and positive predictive values for RAC and BD at 18, 20, 22, 24 and 28 years old for both genders. Background: Alcohol dependence (AD) is a severe disorder Conclusions: Te perception of a freshmen college student accompanied by mental and physical health problems. Te about the amount of alcohol they consume, is an important complex pathogenesis includes environmental and genetic

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 389 Congrès factors. Evidence is emerging that epigenetic mechanisms tionnellement associée à la notion de plaisir et de convivialité might contribute to gene environment interactions which dans tous les milieux. seem to play a major role in the manifestation of addiction. In Cependant, les représentations sociales portant sur les “alcoo- previous studies, we already identifed several genes as being liques” ont toujours été paradoxales et ambivalentes. L’une diferentially methylated in patients compared to healthy des stigmatisations la plus “populaire” aujourd’hui est l’idée controls. Interestingly, our top hits are both involved in the générale que les personnes soufrant d’alcoolisme ne sont pas cellular stress response and in neurodevelopment. malades, mais sont responsables de leur sort. Relativement One of the biggest challenges in psychiatric epigenetics is aux personnes présentant d’autres maladies mentales, celles the inaccessibility of living brain tissue. Terefore, in this dépendantes de l’alcool sont fortement rejetées et soufrent study we tried to replicate our previous hits, which had been de stéréotypes négatifs. identifed in human blood, in human post mortem brain Une source importante de stigmatisation et de discrimination samples as well as in blood and brain samples derived from est trouvée par certains, dans le personnel de santé lui-même a rat model for AD. (incluant les infrmier(e)s), considéré comme en étant le pre- Methods: We investigated post mortem human brain samples mier contributeur. Pour mieux comprendre et identifer les originating from Brodman area 9 either from AD patients déterminants de la stigmatisation dans le Trouble de l’usage (n=13) or healthy controls (n=10). In addition, brain and de l’alcool (TUA), nous avons efectué deux études entre blood samples from a rat model of AD were investigated. novembre 2018 et juin 2019 afn de décrire les représenta- To induce AD, rats had been exposed to daily intermittent tions socio-culturelles et le stigma associés au TUA chez 24 cycles of alcohol vapor intoxication and withdrawal. Rats patients accueillis au Centre hospitalier Esquirol, et leurs were weight-matched and assigned to two groups which were proches, et chez 594 professionnels travaillant dans le même either exposed to ethanol vapor (n=16) or normal air (n=16). hôpital. Nous avons utilisé le questionnaire Explanatory Samples from blood and from diferent brain regions of the Model Interview catalogue (EMIC) qui est un outil d’entretien same animal were obtained one day after alcohol abstinence. semi-structuré permettant une évaluation quantitative et qua- DNA methylation levels in our top hits were assessed by litative des représentations de la maladie, des connaissances, pyrosequencing. du stigma associé et de la recherche d’aide. Results: Interestingly, for the human brain samples both Les résultats montrent que 36 % des patients et 43 % des genes showed diferential DNA methylation when compa- proches ne connaissent pas la maladie, et les différents ring patients with healthy controls. However, for both genes types d’aide disponibles ne sont pas connus pour plus de the changes in methylation were in opposite direction to our la moitié. L’analyse du discours des malades et des proches previous results comparing human whole blood samples. In montre la présence d’une stigmatisation importante envers the rat model, we found diferential DNA methylation levels les personnes soufrant de TUA. Par ailleurs, l’enquête chez for both genes in the PrLC. les professionnels hospitaliers montre une expression plus Discussion: Our results further facilitate the potential role faible du stigma chez les personnes travaillant au sein du of the two investigated candidate genes in AD. Interestingly, Pôle universitaire d’addictologie (PUAL). Ceci met en avant both genes play a role in the cellular stress response as well l’importance de la formation sur la maladie, ou du moins d’un as in neurodevelopment. However, more research is needed contact plus important avec les malades pour la diminution to examine on a cellular level how the regulation of these de la stigmatisation de ces derniers. two genes might be involved in AD. Furthermore, in the rat Les actions de prévention visant à mieux identifer les sources brain we found diferential methylation patterns in the PrLC, d’aide aux patients et familles doivent être entreprises. La a brain region thought to be involved in drug-seeking beha- formation addictologique apparaît indispensable auprès des vior. Our results further emphasize the tissue and cell type professionnels pour mieux combattre la stigmatisation. specifcity of epigenetic changes. Although we can sometimes assume a correlation of DNA methylation changes between Sociocultural representations and stigmatization diferent cell types, further studies are needed to identify related to alcohol use disorder: The perspective which genes do correlate between certain tissue and cell types of patients, their relatives, and professionals at a and which seem to be regulated independently. University addiction center in Limoges (France) Thibaut Dumontheil1, Jean-Jacques Yonga1, Murielle Girard1, Représentations socioculturelles et stigmatisation Philippe Nubukpo1 (1 Limoges, France) liée au Trouble de l’usage d’alcool : le point de vue de patients, de leurs proches et In France, the use of alcohol is very frequent, and is placed de professionnels d’un Pôle universitaire in a context of a cultural habit. Tis psychoactive substance d’addictologie à Limoges (France) is traditionally associated with the notion of pleasure and Thibaut Dumontheil1, Jean-Jacques Yonga1, Murielle Girard1, conviviality in all environments. 1 1 Philippe Nubukpo ( Limoges, France) However, social representations of “alcoholics” have always been paradoxical and ambivalent. One of the most “popular” En France, l’usage d’alcool est très fréquent, inscrit dans une stigmas today is the general idea that people with alcoholism habitude culturelle. Cette substance psychoactive est tradi- are not sick, but are responsible for their trouble. Relative to

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 390 Congrès people with other mental disorders, those dependent on alco- independent of alcohol consumption status (relapse, absti- hol are strongly rejected and sufer from negative stereotypes. nence). Te results allow an overview of some dysfunctions at An important source of stigma and discrimination is found specifc times of the disorder, and refect the characteristics of in the care teams tehmselves (including nurses), which are the problematic alcohol use and not those from the addictive considered to be the frst contributors. To better understand component of AUD. and identify the determinants of Alcohol Use Disorder Source of funding: Inter-regional Clinical Research Hospital (AUD), we conducted two studies between November 2018 Program 2011 - Esquirol Hospital Center. and June 2019 to describe the socio-cultural representations and stigma associated with AUD at the Esquirol Hospital Impact of addictology expert patients on medical Center in Limoges (France) with 24 patients and their rela- students: A qualitative study tives, and 594 professionals working in the same hospital. We C. Obadia1, A. Nguyen Van Nhieu1, D. Moisan2, M. Claudon1, used the “Explanatory Model Interview Catalog” (EMIC) M. Lejoyeux1, A. Reyre3 (1 Paris, France, 2 Clichy, France, 3 Bobigny, questionnaire which is a semi-structured interview tool France) allowing quantitative and qualitative evaluation of the repre- sentations of illness, the knowledges, the associated stigma Background: Te expert patient (EP) participation in medi- and the help seeking. cal education is growing in France. Tey are involved in the Te results show that 36% of patients and 43% of relatives topic of addictions, but the literature is poor on this subject. do not know the disease. The analysis of the speech of However, excessive alcohol consumption is the second lea- patients and relatives shows the presence of a signifcant ding cause of preventable death in France and screening for stigma towards people sufering from AUD. In addition, the alcohol use disorders is one of the least common. Tis study survey of hospital professionals shows a weaker expression evaluates the impact of EPs’ intervention on medical students. of stigma among people working in the University Service Design: It’s a qualitative study. Semi-directed interviews were of Addictology in comparison with the other health care conducted with 11 students, 4 weeks and 4 months after an workers. Tis highlights the importance of formation on addiction training involving 4, conducted in October 2017. the disease for care workers, or at least greater contact with Te analysis of transcribed data follows the thematic method. patients to reduce the stigma of the latter. Findings: Te study identifes 3 dimensions along which Prevention actions to better identify sources of support for students express themselves in diferent postures: the student patients and families should be undertaken. Te training on appreciates the innovative, enriching and playful nature of AUD for the professionals appears essential to better fght this training, through the philanthropic contribution that the stigmatization. seems to be missing in the usual curriculum. Tey unders- tand and retain better. Tey project themselves more into Clinical and biological monitoring of subjects with their clinical practice. Tey also adopt a citizen’s posture by Alcohol Use Disorder after alcohol withdrawal seeing their representations of dependent individuals upset, treatment which leads to a questioning of their relationship with others. Murielle Girard1, Marine Pareaud1, Paul Carrier1, Tese refections lead the physician to question his own way Philippe Nubukpo1 (1 Limoges, France) of practicing medicine and to improve his clinical practice. Conclusions: Tis study reveals the interest of a collective Te diagnosis and follow-up of the subjects with alcohol refection about the contribution of EPs during medical edu- use disorder (AUD) are based on clinical examination and cation, and then about the pedagogical models of the medical biological measure of alcohol use toxicity indicators. Te studies. Prospectively, this work makes it possible to consider rigorous examination of the symptomatology corresponds the integration of EPs into medical education concerning to a moment of the expression of the disorder, and does not taboo subjects or even more globally in medical education. make it possible to predict the success of the treatment, or to Alcohol-Related Brain Injury in alcohol use make an evolutionary prognosis, or characterize dependance. disorder patients attending a UK secondary care Biological indicators could help to objectify them. We sum- hospital marize here the contributions of diferent studies that we 1 1 1 carried out in a 6-month follow-up of a cohort of subjects Lynn Owens , Andrew Thompson , Munir Pirmohamed , 1 1 with AUD who came for alcohol withdrawal at the psychia- Paul Richardson ( Liverpool, United Kingdom) tric hospital. Follow-ups at 1, 2, 4 and 6 months after alcohol withdrawal focused on clinical indicators (relapses, comor- Introduction: Alcohol-Related Brain Injury (ARBI) is an bidities, depression, anxiety, craving), and specifc biological umbrella term for a number of neuropsychiatric conditions indicators : liver stifness, and serum levels of molecules of caused by headrinking. Tis condition is under-recognised interest in the pathophysiology and expression of AUD: and under-treated, often resulting in readmissions leading to neuronal plasticity indicator (Brain Derived Neurotrophic increased physiological and psychological harm for patients Factor), pro-infammatory cytokines, and neuronal damage and relatives. markers (S100 beta and Neuron Specifc Enolase). Te varia- Methods: A 12-month cohort study on the implementation tions of these indicators indicate a long-term improvement of the Montreal Cognitive Assessment (MoCA) tool for

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 391 Congrès detection of ARBI in heavy drinkers. Primary measure of in- disorder for SHAS-E-great, and neurological-disorder for terest was MoCA ≤23. Participants were all in-patients aged SHAS-E-terrible. Enriched networks in pleiotropic variants ≥18 years who were reviewed by the Alcohol Care Team’s included fatty-acid-metabolic-process for SHAS-E-total, Specialist Nurses (n=1276), and need for MoCA screening neurotrophin-signaling-pathway for SHAS-E-great and was based on pre-determined criteria. -terrible. Results: In 12 months, 205 patients were screened, 38% were Our study identifed a glutathione metabolic gene to be sug- initiated due to concerns about cognition raised by relatives. gestively associated with SHAS-E-total in AI, and suggests Of those screened, the directly standardised period prevalence that the genetic substrates underlying LR might relate to cell rate for MoCA ≤23 was 36.1% (n=74). Drinking measures adhesion, neurotrophin, and infammatory responses. were not predictive of either; the need for screening or the Supported by: K25AA025095, AA010201, AA027316, presence of ARBI. Te most common (27%) co-morbidity DA030976. was Alcohol Related Liver Disease (ARLD). In patients with a MoCA ≤23 at 12-month follow-up, 17.5% had died Comparing non-treatment-seeking participants of which 61.5% had no evidence of ARLD. Mean hospital in alcohol medication research studies across attendances were signifcantly reduced from 8 to 5.6 (95%CI: demographic and clinical variables by geographic 1.1 to 6.4; P=0.08) as were admissions from 3.2 to 2.4 (95% location CI: 0 to 1; P=0.03). Roberta Perciballi1, Bianca Persaud1, Victoria Long1, Conclusions: Screening for ARBI in acute hospital settings is Daria Piacentino1, Xiaobai Li1, Lorenzo Leggio1, an important frst step in improving identifcation and mana- Carolina Haass-Koffler1 (1 Providence, USA, Rome, Italy) gement of this patient group with complex medical histories. Further work is required to optimise screening processes and Background: Pharmacotherapy development for Alcohol to develop appropriate treatment pathways. Use Disorder (AUD) treatment is a long and challenging process. One of the challenges in early stage clinical research Assessing the genetic aspects of subjective level for AUD medication is that typically the enrolled partici- of response to alcohol in an American Indian pants are heavy drinkers, but they are not seeking treatment population through whole genome association for AUD. Tis may impact the translational efort to move and pleiotropic analyses medications from clinical research to a clinical setting. One Q. Peng1, K.C. Wilhelmsen2, C.L. Ehlers1 (1 La Jolla, USA, 2 Chapel study found various signifcant diferences between non- Hill, USA) treatment-seeking participants with AUD from human laboratory studies conducted at the University of California, Subjective level of response (LR) to alcohol is considered Los Angeles (UCLA) when compared to treatment-seeking a risk factor for alcohol use disorders (AUD). LR varies by participants with AUD from a national, multi-site clinical ethnicity. Previous studies showed that American Indians trial, the COMBINE study, which evaluated naltrexone and (AI) were less sensitive to the alcohol efects, which could acamprosate with or without a combined behavioral interven- partially contribute to their elevated AUD rates. In this study, tion (CBI). Te UCLA studies were confned to one homo- we assessed how LR related to AI ancestry and AUD severity, genous geographic location, which showed signifcantly less conducted GWAS and pleiotropic study of LR measures in variability between participants compared to the COMBINE 684 admixed AI participants. Tree LR traits from Subjective study, which was conducted at 11 sites across the country. High Assessment Scale Expectancy for alcohol (SHAS-E) Objectives: This present analysis aims to compare non- were evaluated: SHAS-E-total, -great, -terrible. AUD seve- treatment-seeking individuals who participated in human rity was derived from SSAGA interview. laboratory studies for medication development at Brown SHAS-E-total was anti-correlated with AI ancestry (r=-0.10, University to non-treatment-seeking individuals that parti- p-value=9.5E-3) and AUD severity (r=-0.16, p=3.3E-5). cipated in similar studies at UCLA across demographic and GWAS identified several revealing suggestive findings: clinical variables. Rs547109 on GCLC was negatively associated with SHAS- Methods: Participants from the Brown University group E-total (p=3.6E-7). Rs547109 has MAF 26% in general (n=240) were compared to participants from the UCLA population, but 41% in AI. GCLC belongs to glutathione (n=213) group. All participants from all studies were non- pathway crucial to ethanol detoxifcation. CACNA1B variant treatment-seeking individuals who met DSM-IV criteria was associated with SHAS-E-great (p=1.2E-7). Variants for AUD and were enrolled in a medication development on SCML4, PLD5, GADL1, PCDH7 were associated trial. The Brown studies were compared across multiple with SHAS-E-terrible (p=6.3E-8 - 4.3E-7). Top pathways demographic, clinical, and alcohol-related variables to the included adherens-junction for SHAS-E-total, toll-like- UCLA studies. All studies at both sites gave participants receptor and neurotrophin signalings for SHAS-E-terrible, same baseline questionnaires and assessments that were used immune-response-signaling for SHAS-E-great. 217-232 for comparison in this analysis. pleiotropic SNPs were selected from GWAS Catalog. Among Results: Analysis comparing non-treatment-seeking partici- these, infammatory-measurement was the most enriched for pants from the Brown studies to the participants from the SHAS-E-total, immune-system-disorder and neurological- UCLA studies revealed signifcant diferences between the

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 392 Congrès populations across demographic and alcohol-related variables. related with cotinine, are involved in stress systems. Tus, Participants in the Brown studies were older (p<0.0001), had smoking may afect stress responses resulting in a possible fewer years of education (p=0.005), and were more likely to be impairment of homeostasis and dysregulation of the central in a committed relationship or previously married (p<0.0001) feedback response. when compared to the UCLA population. Participants in the Brown studies also had less alcohol dependence (p=0.01) and Relationships between hepatic function, cognitive consumed fewer drinks 30 days prior to their baseline session status and cerebral macrostructure in Alcohol Use (p<0.05) than the participants in the UCLA studies. Disorder patients (AUD) Conclusions: Signifcant diferences between the non-treat- A.L .Pitel1, A. Lanquetin1, S. Segobin1, S. Wilson1, A. Laniepce1, ment-seekers within diferent geographic locations raises F. Vabret1, N. Cabé1, L. Coulbault1, F. Eustache1, D. Vivien1, further questions and the need for more research to further M. Rubio1 (1 Caen, France) defne AUD populations. Future directions will compare non-treatment-seekers in AUD medication studies at Brown Introduction: Alcohol Use Disorder (AUD) has been associa- University to the treatment-seekers in the COMBINE study. ted with cognitive impairments, brain macrostructure altera- tions and liver diseases but their relationship remains unclear. Effects of smoking on endogenous hormones in Moreover, their inherent pathophysiological mechanisms are individuals with alcohol use disorder still unknown. Some studies found no relationship between 1 1 1 Bianca Persaud , Roberta Perciballi , Lorenzo Leggio , biological markers of liver function and structural brain 1 1 Carolina Haass-Koffler ( Providence, USA) abnormalities while others reported correlations between in- creased GGT levels and both gray and white matter atrophy. Introduction: Among individuals with alcohol use disorder GGT level is not a specifc measure of alcoholic liver disease, (AUD), smoking is prevalent. Smoking has multiple efects unlike liver fbrosis (estimated by Fibrometre®) which has, on hormone secretion, some of which are associated with however, never been compared to structural brain alterations important clinical implications. Cotinine represents the most or cognitive impairment. Our aim was to explore the rela- reliable biomarker of smoking behavior. Some signifcant tionships between liver function and brain structure and co- correlations exist between this biomarker and endogenous gnitive function in AUD patients recently after detoxifcation. hormones. Beta-endorphins, is an endogenous opioid neu- Materials and methods: Thirty-two recently detoxified ropeptide known to be involved in stress responses and AUD patients and 20 healthy controls (HC) underwent a maintain homeostasis, released in the peripheral circulation neuropsychological evaluation, a volumetric T1-weighted are afected by nicotine stimulation; nicotine can afect beta- MRI examination and blood sample tests. First, between- endorphins concentration resulting in changes in pain thres- group comparisons were conducted between AUD and HC hold and immune response. , a pineal hormone, on the neuropsychological scores, measures of GGT levels, exerts potential efects on smoking induced oxidative stress. fbrosis scale and gray matter volumes using voxel-based Plus, melatonin can help to counteract the acute efects of morphometry (SPM12). Second, to establish the rela- smoking cessation on mood. Alpha-Melanocyte-stimulating tionship between these measures, correlations were carried hormone (alpha-MSH) an endogenous peptide hormone out between hepatic enzyme levels and liver fbrosis scales, and neuropeptide of the melanocortin family, production is gray matter volumes and cognitive performance. stimulated by nicotine. Substance P neuropeptide, represents Results: Te neuropsychological evaluation showed lower a key responder to preserve biological integrity, likely resul- results in AUD patients than in HC in executive functions, ting in stress responses. Oxytocin is a hypothalamic peptide balance and working memory. AUD patients presented de- hormone and a neuropeptide. It is involved in empathy, sexual creased gray matter volume in the frontal cortex, cerebellum, reproduction and childbirth. Smoking indirectly modulates cingulate and limbic structures including the thalamus and oxytocin neuronal activity determining changes in environ- the hippocampus (FWE, p<0.05, k=100). Compared to HC, mental and stress responses. Also, nicotine can afect AUD patients presented GGT levels and mild liver fbrosis. activity, increasing appetite. Orexin regulates the release of Liver fbrosis negatively correlated with motor abilities of the noradrenaline, one of the neurotransmitters involved in stress. upper limb and the putamen volume. GGT levels negatively Orexin also is implicated in the induction of behavioral res- correlated with gray matter volumes in the left middle frontal ponse to stressors. gyrus and the right temporal gyrus, as well as with cerebellar Method: We performed a pilot study with 18 patients with ataxia. alcohol use disorder to investigate the relationship between Conclusion: First, our results confirm previous findings cotinine and endogenous hormones. regarding GGT levels, liver fbrosis, gray matter abnormali- Results: We found strong positive correlations between ties and neuropsychological defcits in AUD patients. To our cotinine and the following hormones: Beta-endorphin knowledge, this is the frst study describing the profle of gray (r16=0.604; p=0.008), melatonin (r16=0.509; p=0.031), alpha- matter and neuropsychological alterations in AUD patients MSH (r13=0.645; p=0.009), Substance P (r16=0.470; p=0.049), with mild liver fbrosis. We fnd a correlation between liver oxytocin (r16=0.667; p =0.002), orexin (r14=0.742; p=0.001). alteration (GGT levels or fbrosis) and global motor impair- Conclusion: We found that most of these hormones, cor- ment without associated neuropsychological alterations. Our

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 393 Congrès data also reveal a link between the level of GGT in serum of alcohol use disorders. Te Brief Young Adult Alcohol and alteration of some brain regions presenting with a par- Consequences Questionnaire (BYAACQ) is often utilised to ticular sensitivity. Taken together, the two diferent profles explore adverse alcohol-related outcomes among tertiary stu- of gray matter alterations observed between GGT levels and dents. Alcohol consumption behaviour assessed via retrospec- hepatic fbrosis may refect two diferent pathophysiological tive summary measures has been linked to BYAACQ score. mechanisms. While hepatic fbrosis might be caused by oxi- It is unclear, however, how drinking assessed in real-time, dative stress in the liver, increased GGT levels might refect in conjunction with variables such as age of drinking onset oxidative stress not only in the liver, but also in the brain as and other drug use might predict severity of adverse alcohol a result of chronic alcohol consumption. Neuropsychological consequences as captured by the BYAACQ. and neuroimaging correlations with liver impairment both Methods: Te psychometric properties of the BYAACQ support the brain-liver axis role in AUD. were explored using a large Australian sample of tertiary students (N=893). A subsample (n=504) provided alcohol Acute IP administration of N-acetylcysteine intake information in real-time (21 days) via a smartphone prevents the activation of the mesocorticolimbic app (CNLab-A) plus age of drinking onset, drug use, and system triggered by intra-VTA ethanol parental alcohol/drug use details. administration in rats Results: Average BYAACQ score for the full sample was S. Fernández-Rodríguez1, C. Esposito1, L. Granero1, A. Polache1, 7.23 (SD=5.47). Classical and item response theory ana- T. Zornoza1, M.J. Cano-Cebrián1 (1 Valencia, Spain) lyses revealed some inconsistencies related to progressive item severity and male/female diferential item functioning. Several studies have explored the potential efcacy of pro- Current drinking – namely, frequency of intake and quantity longed N-acetylcysteine (NAC) treatments, to regulate/ per drinking occasion – plus age of drinking onset and other modify ethanol-related behaviours. Its effects has been drug use accounted for 33.9% of the variance in BYAACQ attributed, at least in part, to the ability of prolonged NAC score after controlling for age and depression symptomology. treatment to reverse ethanol induced plasticity. In this sense, Conclusions: Information related to current drinking, age of one of the most widely accepted hypotheses assumes that drinking onset, and drug use is useful for predicting severity chronic NAC treatment is able to restore the expression of of alcohol use consequences. Tese markers might enable glial transporters, such as xCT and GLT-1, contributing to tertiary institutions to target students for prevention/inter- the normalization of the glutamate function in the striatal vention programs. system. However, on the other hand, recent research has also shown that an acute dose of NAC is enough to limit DNA Methylation Analysis of the GABRA2 receptor motivation, seeking behaviour and reacquisition after ethanol subunit in alcohol dependence self-administration in rats. In order to give a plausible expla- Ulrich W. Preuss1,2, Gabriele Koller3, Peter Zill3 (1 Halle/Saale, nation to these latest results, in the present work, we have Germany, 2 Herborn, Germany, 3 Munich, Germany) explored whether NAC is able to counteract the activation of mesocorticolimbic system triggered by acute intra-VTA ethanol administration. To this end, we combined local Background: Variants of GABRA2 have been repeatedly administration of 150 mM of ethanol in the posterior Ventral associated with alcohol dependence risk. However, no study Tegmental Area (pVTA), with the systemic administration investigated potential epigenetic changes in GABRA2 CpGs (IP) of NAC (60 or 120 mg/Kg) 30 min before. Afterwards, between alcohol-dependent (AD) subjects and controls and we measured the neuron cFOS inmunoreactivity in the relationship to AD characteristics. Nucleus Accumbens by immunohistochemical analysis. Our Methods: In the present study, blood samples for promoter- results show that 120 mg/kg of NAC, but not 60 mg/kg, pre- related GABRA2 CpG and genome wide global methylation vented the increment in the cFOS immunoreactivity induced were obtained from n=57 AD subjects and 51 controls which by ethanol. In this scenario, an alternative mechanism of were clinically assessed by structured interviews. Global action of NAC, apart from the above mentioned, cannot be 5-methylcytosin (5-mC) methylation was measured using ruled out. Terefore, further experiments are necessary to ELISA kits for 5-mC. Assessment of random DNA methy- elucidate the mechanism of action of NAC. lation status was accomplished using the [3H] methyl group incorporation assay. Real-time drinking, age of drinking onset, and Results: While no signifcant diference in GM was detected other drug use predict adverse alcohol use across groups, after controlling for age and gender, mea- consequences of tertiary students sures of GABRA2 epigenetic changes yielded a signifcant Antoinette Poulton1, Adrienn Mata1, Jason Pan1, hypomethylation of several CpG sites. Hypomethylation Loren Richard Bruns1, Richard O. Sinnott1, Robert Hester1 was related to recent alcohol intake, duration of AD and (1 Parkville, Australia) withdrawal severity. Summary: Te results indicate a signifcant epigenetic change Background: Compared to their non-university enrolled in GABRA2 methylation which is also related to AD seve- peers, tertiary students consume greater quantities of alcohol, rity. Further studies are needed to determine the efects of are at increased risk of injury/harm, and have higher rates epigenetic changes on GABRA2 expression and longitudi-

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 394 Congrès

8 - Drummond C, Wolstenholme A, Deluca P, Davey Z, Donoghue K, Elzerbi nal changes of epigenetic regulation over time to clarify the C, et al. Alcohol policy in Europe: Evidence from AMPHORA. 2013. pathophysiological relevance of the fndings. 9 - Anderson P, O’Donnell A, Kaner E. Managing Alcohol Use Disorder in Primary Health Care. Curr Psychiatry Rep. 2017 ; 19 (11) : 79. In search of perfection: defining new approaches 10 - World Health Organization. WHO Collaborative Project on to improve the EIBI-culture in general practice Identification and Management of Alcohol-Related Problems in Primary Health Care: report on phase IV: development of country-wide strategies 1 1 Bram Pussig ( Leuven, Belgium) for implementing early identification and brief intervention in primary health care. Heather N, editor. 2006. 11 - Keurhorst M, Heinen M, Colom J, Linderoth C, Müssener U, Okulicz- Background. Early Identifcation and Brief Interventions Kozaryn K, et al. Strategies in primary healthcare to implement early (EIBI) in primary care is a proven (cost-)efective method identification of risky alcohol consumption: why do they work or not? A for reducing harmful drinking in society (1-5). Unfortunately, qualitative evaluation of the ODHIN study. BMC Fam Pract. 2016 ; 17 : 70. integration in daily practice is low (6-8). It is suggested that 12 - Seppä K. Development of country wide startegies for implementing early identification and brief intervention in primary care. Report on Phase a more integrated EIBI-culture is necessary to fully utilise its IV of WHO collaborative project. Nord alkohol Nark. 2003 ; 20 : 91-6. efect (9-11) and that community involvement might provide a tool to stimulate that (10, 12). Te still suggestiveness of Role of estrogen in the gender effect of binge- this approach makes the stakeholders’ point of view essential drinking on hippocampus LTD in young adult rats for defning community-oriented actions to stimulate the K. Rabiant1, J. Antol1, M. Naassila1, O. Pierrefiche1 (1 Amiens) negotiability of alcohol use in primary care; creating a bene- fcial setting for EIBI delivery. Aim. Te aim of this study is to defne community-oriented Binge-drinking is responsible for memory impairments espe- strategies to stimulate the negotiability of alcohol use in cially during adolescence. Recent studies highlighted a higher general practice. sensitivity of women brain regarding the efects of ethanol. Method. Stakeholders will be asked to defne community- However, the reason behind such diference remains unclear. oriented strategies to normalise the negotiability of alcohol Here, we tested the hypothesis that estrogen level (E2) play use in general practice. Te nominal group technique will a role in the gender diference observed in the efects of be applied, it allows generating ideas within a stakeholder ethanol on hippocampus plasticity, the cellular mechanisms population while creating consensus. Forty-eight stakeholders implicated in learning and memory processes. Long-Term from the municipality of Leuven will be divided into four he- Depression (LTD) was recorded in hippocampus slices 24 h terogeneous nominal group sessions. Tese sessions combine after 2 binge-like ethanol exposure (3 g/kg, i.p.) applied individual and group work that comprises generating ideas, during the diferent phases of the oestrus cycle. We also used sharing ideas, discussing ideas and voting on ideas; resulting exogenous E2 (180 µg/kg) with and without concomitant in immediate action planning. All results will be merged presence of ethanol during specifc phases of the oestrus cycle into an overarching list, based on validated implementation in postpubertal rats and prepubertal female rats. We found frameworks. A member check session will be conducted to that neither oestrus cycle nor ethanol binge alone had an ensure correct interpretation of the results. efect on LTD magnitude in postpubertal female rats at 24 h Results. A prioritised list of scientifc robust and stakeholder- delay. However, LTD was abolished at 24 h delay only when inspired strategies, for stimulating the negotiability of alcohol ethanol was injected during the endogenous peak of E2. Such use in general practice, is generated. Developed community- abolition of LTD was also obtained when co-injection of E2 oriented actions will be presented at the conference. and ethanol was performed in either postpubertal female rats References in low E2 phase or prepubertal rats. Finally, we measured a similar abolition of LTD in male rats at 24 h delay when 1 - Angus C, Scafato E, Ghirini S, Torbica A, Ferre F, Struzzo P, et al. Cost- effectiveness of a programme of screening and brief interventions for alco- ethanol was injected with a higher dose of ethanol (3.75 g/kg, hol in primary care in Italy. BMC Fam Pract. BioMed Central. 2014 ; 15 : 26. i.p.). Our results showed that for the same dose of ethanol, 2 - Kaner EF, Beyer FR, Muirhead C, Campbell F, Pienaar ED, Bertholet N, et LTD in female rats is more sensitive than in males, especially al. Effectiveness of brief alcohol interventions in primary care populations. Cochrane Database Syst Rev. 2018 ; 2 : CD004148. when ethanol was present at the same time that a high level 3 - O’Donnell A, Anderson P, Newbury-Birch D, Schulte B, Schmidt C, of estrogen. Tese results suggests that E2 plays a role in the Reimer J, et al. The impact of brief alcohol interventions in primary health- gender diference of ethanol efects in hippocampus plasticity. care: a systematic review of reviews. Alcohol Alcohol. 2014 ; 49 (1) : 66-78. 4 - Purshouse RC, Brennan A, Rafia R, Latimer NR, Archer RJ, Angus CR, et Alcohol consumption and gender gap in al. Modelling the cost-effectiveness of alcohol screening and brief interven- tions in primary care in England. Alcohol Alcohol. 2013 ; 48 (2) : 180-8. cardiovascular mortality in Europe 5 - Angus C, Latimer N, Preston L, Li J, Purshouse R. What are the Razvodovsky Yury1 (1 Grodno, Russia) Implications for Policy Makers? A Systematic Review of the Cost- Effectiveness of Screening and Brief Interventions for Alcohol Misuse in Primary Care. Front Psychiatry. 2014 ; 5 : 114. Background: Cardiovascular disease (CVD) is the largest 6 - Bendtsen P, Anderson P, Wojnar M, Newbury-Birch D, Müssener U, contributor to the morbidity and mortality in Europe. Colom J, et al. Professional’s Attitudes Do Not Influence Screening and Mortality from cardiovascular disease remains substantially Brief Interventions Rates for Hazardous and Harmful Drinkers: Results from ODHIN Study. Alcohol Alcohol. 2015 ; 50 (4) : 430-7. higher among men than among women. Te level of alcohol- 7 - Anderson P. Overview of interventions to enhance primary-care provi- related problems difers substantially across Europe, with der management of patients with substance-use disorders. Drug Alcohol Eastern European countries experiencing higher burden of Rev. 2009 ; 28 (5) : 567-74. alcohol-attributable morbidity and mortality than Western

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References European countries. - Leasure JL, Neighbors C, Henderson CE, Young CM. Exercise and alcohol Objective: Tis study aims to test the hypothesis that alcohol consumption: What we know, what we need to know, and why it is impor- plays an important role in explaining the gender gap in CVD tant. Frontiers in Psychiatry. 2015 ; 6 : 156. doi: 10.3389/fpsyt.2015.00156. mortality in Eastern Europe. - Rodríguez-Arias M, Navarrete F, Blanco-Gandia MC, Arenas MC, Bartoll- Methods: Te male-to-female ratio of CVD mortality and Andrés A, Aguilar MA, Rubio G, Miñarro J, Manzanares J. Social defeat in adolescent mice increases vulnerability to alcohol consumption. Addiction the level of alcohol consumption per capita in Western (n 21) Biology. 2016 ; 21 (1) : 87-97. doi:10.1111/adb.12184. and Eastern European (n 24) countries were compared. Te male-to-female ratio of CVD mortality (the fve-year average Incidental neoplasia and mortality among heavy from 2010 to 2014) was calculated. Te comparison in the alcoholics gender gap in CVD mortality was made between Western (n Iván Ribot1, Victor E. Vera Delgado1, Lourdes Gonzalez 21) and Eastern (n 24) European countries (t-test). 1 1 1 Results: Te results of the correlation analysis indicate statis- Navarrete , Paula Reyes Suárez , Ana Godoy Reyes , 1 1 tically signifcant relationship between alcohol consumption Candelaria Martín González , Esther Martín Ponce , 1 1 per capita and gender gap in CVD mortality in Eastern Emilio González Reimers ( Tenerife, Canary Islands, Spain) Europe. Te relationship between alcohol consumption and gender gap in CVD mortality in Western Europe is also We included 255 alcoholic patients (aged 59±11; drinkers of positive, but statistically non-signifcant. 200±150 g/day during 32±12 years), consecutively admitted Conclusion: Alcohol appears to play an important role in the to the Internal Medicine Service of our hospital due to orga- gender gap in CVD mortality in the countries of Eastern nic complications of alcoholism. who were followed up du- Europe. ring a period of 5 years. During this time 103 died. 110 were cirrhotics; 61 of them died during this period, and mortality Physical activity reverses the increase in ethanol was clearly related to liver cirrhosis (Log rank (LR)=9.26; oral self-administration induced by social defeat p=0.002). Thirty nine patients developed neoplasia, 18 in male mice among cirrhotics and 21 among non cirrhotics (x2=0.034; Marina D. Reguilón1, Carmen Ferrer-Pérez1, José Miñarro1, NS). Te development of neoplasia was not related to liver Marta Rodríguez-Arias1 (1 Valencia, Spain) function (assessed by Child-Pugh´s score, LR=2.5; NS), but it was marginally related to the amount of ethanol ingested Preclinical and clinical studies have shown that exposure (LR=2.97; p=0.08). Survival was signifcantly related with to stress increases drug-seeking and relapse into ethanol cancer (LR=5.46; p=0.019), especially among non-cirrhotics (EtOH) consumption by modifying the activity of brain (LR=13.026; p<0.0001), but not among cirrhotics (LR=0.016; areas involved in the rewarding efects of EtOH. In a pre- NS). Te total amount of ethanol ingested was not related vious study we demonstrated that exposure to repeated social to mortality either in the whole group (LR=0.85; NS), non- defeat (RSD), a model of social stress, produced a long-term cirrhotics (LR=0.51), or cirrhotics (LR=2.1), but mortality increase in the consumption of EtOH. Te aim of the present was associated with Child-Pugh´s classifcation (LR=9.81; work was to evaluate if exposure to physical activity can block p=0.007). By Cox regression model the variable cirrhosis the increase in EtOH consumption induced by RSD. Mice entered in the frst place in relation with survival, followed were exposed to 4 sessions of repeated social defeat in which by the variable cancer. 150 patients underwent assessment of they were confronted with an aggressive animal (resident), handgrip strength using a Collins dynamometer. Mortality while the control groups were exposed to a similar situation was clearly related to handgrip strength (LR=8.1; p=0.004). without an aggressive opponent (exploration). During the However, handgrip was displaced by liver function and can- whole procedure, half of the mice were exposed to controlled cer using Cox regression model. Terefore, we conclude that physical activity, being allowed 1 h access to a low-profle neoplasia is a common fnding among heavy alcoholics, and running wheel three times a week. Tree weeks after the last is related to mortality both in the whole group and, especially, social defeat, animals began oral self-administration (SA) among non-cirrhotics. If cirrhosis has already developed, an of ethanol (6% EtOH). Our results show that the socially incidental neoplasia has no efect on mortality. Although defeated animals not exposed to physical activity consumed handgrip stength was associated with mortality in the univa- greater amounts of ethanol and showed greater motivation to riate analysis, it was displaced by liver function and neoplasia obtain the substance than the non-stressed group. Defeated in the Cox regression analysis. animals that performed physical activity behaved similarly to the non-stressed exploration group. Terefore, our results Effects of alcohol hangover on working confirm that controlled physical activity can reverse the memory process in university students – efects of social stress on EtOH consumption. An electroencephalography study Acknowledgements: Ministerio de Economía y Rui Rodrigues1, Natália Antunes1, Eduardo López-Caneda1, Competitividad (MINECO), PSI2017-83023-R; Instituto Adriana Sampaio1, Alberto Crego1 (1 Braga, Portugal) de Salud Carlos III, Red de Trastornos Adictivos (RTA) RD16/0017/0007 and Unión Europea, Fondos FEDER Binge drinking (BD), characterized by an excessive intake “una manera de hacer Europa”; Te European Foundation of alcohol – 5 or more drinks for males and 4 or more for for Alcohol Research (ERAB), EA13 08. females in two hours or less –, leading to elevated blood

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 396 Congrès alcohol concentration (BAC), equal or higher than 0,08 g/dL Liver cirrhosis induces profound immune suppression (cirrhosis- over a brief period followed by abstinence, has acquired associated immune dysfunction, CAID) accounting for in- great attention due to its negative social and health conse- creased susceptibility to bacterial infection and mortality. We quences (car crashes, assaults, low academic performance, have previously shown the “leaky” gut as pivotal in driving cardiovascular diseases), as well as its high prevalence among this phenomenon in Alcohol-related Liver Disease (ALD) adolescents. Tese data, alarming by themselves, become even (Riva et al, Gut 2018; Markwick et al, Gastroenterology more worrying considering that adolescence is an especially 2015), with bacterial translocation due to increased gut per- vulnerable period to the neurotoxic efects of alcohol due to meability stimulating systemic/intrahepatic infammation. the structural and functional changes undergoing in the brain However, the pathways that underlie gut hyper-permeability at this stage. in ALD are not well understood. It is also known that alcohol intoxication has major detri- As part of a “European Foundation for Alcohol Research” mental efects on functioning the following day (e.g., len- (ERAB)-funded study, we performed colon transcriptomics tifcation, difculties in maintaining focus, poor decision in 10 alcohol-cirrhotic patients (ARC) and 10 healthy making). Furthermore, after full metabolism of the alcohol controls (HC) (~20,000 genes, ArrayStar). We investigated consumed, symptoms like headache, diarrhea, tremulousness, differential gene expression and functional clustering by fatigue or nausea may appear. Te presence of two or more “Gene Ontology” (GO), “Kyoto Encyclopaedia of Genes and of these symptoms as the outcome of excessive alcohol intake Genomes” (KEGG) pathways and “Gene Set Enrichment is called hangover. Literature has reported that hangover Analysis” (GSEA). We also measured surrogate markers of can signifcantly decrease alertness and arousal performance bacterial translocation (D-Lactate, Endotoxin) as indicative which can be caused by a decrease in the sleep quality, afec- of gut barrier disruption. ting the ability to react quickly, and psychomotor ability in ~8,400 genes were diferentially expressed in ARC vs HC drive-tasks. However, despite the importance of the imme- colon. Most upregulated genes were functionally involved diate consequences of BD to daily life, few are the studies in transcriptional processes, ribosomal/chromatin structure, that so far have evaluated the short-term efects of a BD intercellular adhesion and energy metabolism. GSEA identi- session on neurocognitive functioning of the adolescent/ fed upregulated adhesion pathways in ARC vs HC, driven by young brain and, in our knowledge, no studies have used tight junction genes including claudin 3, occludin and ZO-1. electrophysiological methods. Conversely, downregulated genes did not cluster signifcantly. Quantifcation of plasma D-Lactate and endotoxin (Lps) Te aim of the present investigation was to assess the beha- indicated the presence of gut barrier disruption and bacterial vioral and electrophysiological consequences on the wor- translocation in ARC vs HC. king memory processes in the real day after a BD session, In conclusion, transcriptional profling highlights major alte- during hangover state, in young people. For that purpose, rations in intestinal cell adhesion pathways in ALD patients. we recorded EEG of 10 university students (6 females) with We are currently investigating these pathways in experimen- a BD consumption pattern while they execute a one-back tal models as new targets to restore barrier integrity in ALD. task, in both a normal and a hangover day. Te reaction times and percentages of correct responses, as well as the latency Don’t stress the amygdala – The role of pro- and and amplitude of P2, N2, P3 and Late Positive Component anti-stress amygdalar systems in alcohol use (LPC) were compared in both moments. disorder (AUD) Despite having found no signifcant diferences at the beha- Marisa Roberto1 (1 La Jolla, USA) vioural level, the hangover was associated with electrophysio- logical anomalies. Te amplitude of ERP components ana- Chronic alcohol exposure in alcohol-preferring (sP) lyzed was smaller in the hangover state. Te lower amplitude rats provokes mild brain and liver inflammatory of early ERP components as P2, N2 and P3 may indicate that responses and a specific atrophy of the corpus during hangover subjects showed difculties to cope as well callosum with the attentional demands of the working memory task in 1 1 1 1 1 regards of the constant updating of information; while smaller M. Rubio , A. Lanquetin , M. Naveau P. Maccioni , I. Lorrai , 1 1 1 1 amplitude of LPC can suggest that subjects show latent def- G. Colombo , D. Vivien , A.L. Pitel ( Caen, France) cits in the ability to recollect or maintain information during the hangover state. Further studies with a larger sample are Introduction: Neuroimaging and neuropsychological studies needed to clarify the detected electrophysiological anomalies. reveal structural and functional brain alterations associated with chronic, excessive alcohol consumption. In 50 to 80% of Gut barrier disruption in alcohol-related liver Alcohol Use Disorder (AUD) patients, these brain alterations disease is characterised by broad alterations of result in cognitive and/or motor impairments. In addition to physiological transcriptional profiles and increased this, 70 to 80% of AUD patients show hepatic damage that bacterial translocation can vary from steatosis to cirrhosis. Te relationship between 1 2 2 1 1 1 hepatic damage and brain dysfunction in AUD is not well A. Riva , J. Bajaj , A. Fagan , R. Williams , S. Chokshi ( London, understood to date and has been mainly investigated within 2 United Kingdom, Richmond, USA) the context of acute hepatic encephalopathy. Here, we have

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 397 Congrès studied macroscopic and microscopic brain and liver altera- assess the neuroimmune component of alcohol-related neuro- tions induced by voluntarily consumed alcohol in selectively toxicity in a binge-like ethanol exposure in adolescent rats bred Sardinian alcohol-preferring (sP) rats, a validated model and ii) to evaluate the impact of nalmefene treatment on of excessive alcohol consumption. alcohol-induced neuroinfammation. Materials and methods: Post mortem brain and liver studies Materials and methods. Adolescent rats (n=6-10 animals/ were performed in sP rats exposed to the homecage, 2-bottle group) received an i.p. injection of ethanol [3 g/kg in 25% “alcohol (10% v/v) vs water” choice regimen with unlimited (v/v)] or saline (control) in a validated intermittent adminis- access for 12 months. Control rats were exposed to 2 water tration pattern (two consecutive days at 48-h intervals over a bottles. Liver infammation was assessed by immunohistoche- 14-day period) (1). In another group, nalmefene (0.4 mg/kg, mical analyses. Brain anatomical measurements were conduc- s.c) was injected 1 hour prior to ethanol. MicroPET imaging ted by T2-weighted MRI scans. Regional brain astrogliosis with [18F]DPA-714 (~37 MBq, i.v.) was performed 24 h after and microgliosis were evaluated by immunohistochemical the last alcohol/saline injection. In each experiment, the brain analysis. distribution of [18F]DPA-714 was estimated in diferent Results: Weekly alcohol intake averaged 35-45 g/kg over the brain areas using the Logan Plot analysis and the metabo- 12-month period. lite-corrected arterial input function. Blood alcohol levels Our results showed mild but significant inflammatory obtained in the model were measured in an independent responses in the liver of alcohol-drinking sP rats, with (i) group using gas chromatography. increased numbers of Kuppfer cells (Iba1+), and (ii) overac- Results. Ethanol administration to adolescent rats induced tivation of hepatic stellate cells (GFAP+). an blood alcohol concentration of 2.40±0.5 g/L at 5 min 18 Concerning the brain, we observed a generalized infammato- after injection. Te regional VTs of [ F]DPA-714 in alco- ry response revealed by increased numbers of microglial cells hol exposed animals (VT hippocampus=21.1±2.7 and VT accumbens= and astrocytes in cortex, corpus callosum, and hippocampus 25.3±6.1) were significantly increased when compared of alcohol-drinking sP rats. Tese infammatory responses to control animals (VT hippocampus=5.9±0.8 and VT accumbens= were accompanied by a specifc atrophy of the corpus callo- 7.07±1.3). Nalmefene signifcantly alleviated the alcohol- 18 sum in alcohol-drinking sP rats, with no changes in hippo- induced increase in [ F]DPA-714 binding (VT hippocampus= campus, cerebellum nor the total brain volume. 14.02±5.22 and VT accumbens=18.5±5.4).Te efects of alcohol Conclusion: We describe here, for the frst time, infammato- and nalmefene were homogeneously observed in all brain ry responses to long-term alcohol drinking in liver and brain areas. of alcohol-preferring sP rats. Tese infammatory responses Discussion. Tese results support the neuroinfammatory could be triggered by increased plasmatic LPS levels, which hypothesis of alcohol-related brain toxicity and suggests that has been recently described in the same set of alcohol-drin- nalmefene may protect from this neuroinfamation. PET king sP rats (Posteraro et al., 2018). Interestingly, the specifc imaging using [18F]DPA-714 is a relevant technique to inves- atrophy of the corpus callosum suggests a vulnerability of tigate the neuroinfammatory component of alcohol exposure this region to chronic alcohol exposure, and is in accordance in animal models and patients. to human studies (Oscar-Berman, 2003). Te absence of Funding : Fondation pour la recherche en alcoologie. anatomical changes in other regions of the brain could, on Reference the contrary, suggest a specifc resistance to alcohol damages. 1 - Pascual M, Montesinos J, Marcos M, et al. Addict Biol. 2017 ; 22 : 1829- Behavioral studies are needed to determine the impact of 41. these macro- and microscopic alcohol-induced alterations. References Dimensionality and Scale Properties of DSM-5 - Posteraro et al. Liver Injury, Endotoxemia, and Their Relationship to Alcohol Use Disorder Intestinal Microbiota Composition in Alcohol-Preferring Rats. Alcohol Clin 1 1 1 Exp Res. 2018 ; 42 (12) : 2313-25. Tulshi D. Saha , Patricia Chou ( Bethesda, USA) - Oscar-Berman, Marinkovic. Alcoholism and the brain: an overview. Alcohol Res Health. 2003 ; 27 (2) : 125-33. Changes to the DSM-5 criteria for alcohol use disorder (AUD) included eliminating the legal problems criterion Nalmefene alleviates alcohol-induced and adding a new criterion, alcohol craving. DSM-IV abuse neuroinflammation: A PET imaging study with and dependence were based on discrete sets of 4 abuse and 7 [18F]DPA-714 in an adolescent rats dependence criteria, but all 11 criteria apply toward DSM-5 W. Saba1, G. Pottier1, M. Goislard1, C. Leroy1, J. Negroni1, mild (2-3 criteria), moderate (4-5 criteria) and severe (6 or S. Demphel1, F. Caille1, C. Coulon1, E. Jaumain1, B. Jego1, more criteria) AUD diagnoses. N. Tournier1 (1 Orsay, France) Using Item Response Teory (IRT), the purpose of this study was to evaluate these cutof points for DSM-5 AUD severity Introduction. Alcohol exposure during adolescence induces using US adult data from NESARC, IRT analyses provided important and long-lasting brain damages, which are assumed test and information functions to examine the diferent level to involve neuroinfammatory processes. [18F]DPA-714 PET of AUD severity and evaluate the interpretation of these imaging targeting the glial biomarker TSPO (Translocator severity level. Protein 18kDa) was performed to i) non-invasively Tere was no signifcant gain of information from DSM-5

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 398 Congrès criteria compared to DSM-IV criteria. Te IRT information associated with greater subjective and behavioral motivation function curve showed that equal ranges in raw scores did not for alcohol. correspond to equal ranges in the latent AUD severity mea- Supported by NIH/NIAAA grants U24 AA014811, sure. For a cutof score of 2+ (mild AUD), the latent severity AA08757, AA025359. level was between 1.2 and 1.6 (a range of 0.7 in logit units), for a cutof score of 4+ (moderate AUD), the latent severity Assessing decision-making impairments in a level was between 1.9 and 2.1 (a range of just 0.2 in logit rodent model of Binge Drinking using relevant units). Tat is, as latent severity increased, the contribution tools: The Rat Gambling Task and ex-vivo of each additional criterion diminished. For cutof score of Fast-Scan Cyclic Voltammetry 6+ (severe AUD), the latent severity level was greater than P. Sauton1, E. Negria1, V. Jeanblanc1, J. Jeanblanc1, M. Naassila1 2.4 in logit units. (1 Amiens, France) Te interpretability and practicality of the DSM-5 AUD severity cut points can be enhanced through the evaluation Decision-making (DM) is an essential cognitive process of IRT test and information functions. resulting in the most advantageous choice among several alternatives. DM has been shown to be altered in alcohol- Long lasting epigenetic marks of alcohol on dependent patients and the few reports in Binge Drinking circadian and stress regulatory genes (BD) subjects are contradictory. Te aim of this study was to Dipak K. Sarkar1 (1 New Brunswick, USA) test whether the BD pattern of consumption is able to alter the DM performances as well and study the dopaminergic Epigenetic modifcations of a gene have been shown to play a correlates within the nucleus accumbens. role in maintaining a long-lasting change in gene expression. In this study, we used a Rat Gambling Task (RGT) paradigm, DNA methylation occurring at CpG dinucleotides is the which is based on the Iowa Gambling Task used in Humans, most common epigenetic modifcation that constitutes an and two models of BD administration: the frst one is an ope- important regulatory element in human genome. Epigenetic rant self-administration model of BD that we developed and alterations believed to occur early in disease state, thus provi- the second is a commonly used intermittent forced adminis- ding the possibility of early diagnosis. As stress and circadian tration of intoxicating doses of ethanol. Secondly, we used ex- physiological systems governing many body functions are vivo fast-scan cyclic voltammetry (FSCV) and dopamine D2 often disregulated in alcohol dependent patients, we sought receptors (D2R) pharmacology in order to identify changes to test whether epigenetic changes of in the dopaminergic transmission in the NAc involved in (Pomc) and period 2 (Per2) genes, critical for stress and cir- these defcits. We show that BD rats made signifcantly less cadian regulation, is long-lasting and may serve as measures advantageous choices than control rats, and that the BD of behavioral motivation for alcohol. To test this, we frst signifcantly increases the proportion of rats with a poor level studied the dose response and time course efects of alcohol of DM. Our FSCV results allowed us to identify a characte- on Per2 and Pomc gene methylation and gene expression in ristic profle of bad DM and the involvement of D2R in the isolated mouse-derive POMC cells in cultures, and found efect of alcohol on the dopaminergic transmission. binge-like ethanol concentrations increase DNA methylation Being able to model the altered DM caused by alcohol and while decrease mRNA expression of Per2 and Pomc genes for fnding relevant therapeutic targets leads to important pers- several days beyond the day of ethanol exposures. In human, pectives in order to study the neurobiological mechanisms we found pregnant women who consumed moderate to high involved in such a deleterious behavior that BD is. levels of alcohol and gave birth to prenatal alcohol exposed Measuring alcohol craving after virtual reality (PAE) children had higher DNA methylation of POMC exposure: A method comparison in social drinkers and PER2. PAE children also had increased methylation of 1 1 1 1 POMC and PER2. In adult humans, non-smoking moderate, Jessica Simon , Léonie Schroder , Etienne Quertemont ( Liège, non-binging compared to binge and heavy alcohol drinkers, Belgium) we found increased methylation of the PER2 and POMC DNA, reduced expression of these genes in the blood samples Craving contributes to the maintenance and relapse of alco- of the binge and heavy drinkers relative to the moderate, non- hol dependence. Some methods, more or less explicit, are binge drinkers. Increased PER2 and POMC DNA methyla- used to estimate this subjective state: on one hand, single tion was also signifcantly predictive of both increased levels items or validated questionnaires and on the other hand, ad- of subjective alcohol craving immediately following imagery, libitum taste test. In this second paradigm, participants are and with presentation of the alcohol (2 beers) prior to the invited to taste and evaluate the organoleptic properties of alcohol taste test, as well as with alcohol amount consumed several alcoholic beverages. Te total amount of liquid drunk during the alcohol taste test. Tese data establish signifcant is an indirect measure of craving. Te objective of the present association between binge or heavy levels of alcohol drinking study is to compare self-reported measures of craving with and elevated levels of methylation and reduced levels of measurements from the ad-libitum tasting test. We hypothe- expression of POMC and PER2 genes. Furthermore, elevated size a relationship between the implicit and explicit measures methylation of POMC and PER2 genes is long-lasting and is of craving. In addition, we will attempt to determine the most

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 399 Congrès valid measure of craving. To do this, we will determine which Prenatal alcohol exposure (PAE) is known to damage the measure is most strongly correlated with the AUDIT score fetal brain and leads to life-long cognitive and behavioral and the obsessive compulsive drinking scale score. 46 social dysfunctions. Fetal Alcohol Spectrum Disorders (FASD), drinkers will be recruited for this experiment and immersed which collectively describes the constellation of effects in a virtual environment including alcohol-related cues, sup- resulting from alcohol consumption during pregnancy, is a posed to generate craving, before evaluating it with single complex syndrome that afects up to 5% of children and is the items and ad-libitum taste test. leading cause of preventable intellectual disability. Despite prevention campaigns discouraging alcohol drinking during Impact of chronic alcohol consumption and pregnancy, the number of children sufering from FASD withdrawal on hippocampus or striatum-dependent has not decreased over the past years. Te consequences learning and related synaptic plasticity of PAE have become a global public health problem and Léa Tochon1, Rose-Marie Vouimba1, Jean-Louis Guillou1, understanding the alcohol-related mechanisms is crucially Daniel Béracochéa1, Vincent David1 (1 Bordeaux, France) needed to develop new pharmacological strategies and treat- ments. Studies have shown that alcohol interferes with the cerebral cortex development in a variety of ways, including Te hippocampus and striatum have dissociable roles in defects in neurogenesis, impaired cell proliferation and cell memory: while the former is necessary for spatial/declara- migration, reduced survival and disrupted neurotransmis- tive forms of learning, the latter underlies cued/procedural sion. However, the precise pathophysiological mechanisms learning. An emerging hypothesis suggests that drug addic- underlying alcohol’s actions on cortical development are yet tion could lead to a functional cognitive imbalance, which poorly understood. In this study, we set up a mouse model would maintain addictive behaviour and support the risk of FASD, using an alcohol consumption paradigm in which of relapse by promoting habit learning while concurrently mice voluntarily drink high amounts of alcohol throughout disrupting spatial memory. We examined, in C57BL/6J male pregnancy. Importantly, this model avoids any bias resulting mice, whether chronic ethanol consumption (5 months) or from maternal stress that could be introduced by stressful withdrawal might modulate the use of spatial memory vs alcohol consumption procedures such as gavage or injection. cued memory, and related hippocampal and striatal synaptic We frst showed that this model accurately refects alcohol plasticity. Using a competition protocol in the Barnes maze consumption in human, as mice reach blood alcohol concen- assessing the respective use of hippocampus vs striatum- tration levels comparable to those reported in binge-drinking dependent learning strategies, we frst show that alcohol humans. In order to investigate alcohol-dependent cortico- withdrawal, and also to a lesser extent alcoholization, dras- genesis defects, we are analyzing the number, proliferation tically increase the use of non-fexible, striatum-dependent and specifcation of glutamatergic projection neurons during cued strategies in a task solved with spatial memory in 95% embryonic development and at postnatal stages. By using of non-alcohol exposed mice. Concurrently, by performing in utero electroporation, we are investigating the migration in vivo electrophysiological studies in freely-moving mice pattern of projection neurons during neurogenesis. Our pre- to assess learning-induced synaptic plasticity in both the liminary results reveal an abnormal accumulation of neurons dorsal hippocampus (CA1) and dorsolateral striatum (DLS), in deep layers of the cortex of alcohol-exposed embryos, we found that task-induced synaptic plasticity activity was suggesting impaired neuronal migration or dysregulated reduced in the CA1 and increased in the DLS of withdrawn layer specifcation. Analysis of radial migration at postnatal mice, and to a lesser extent, in alcohol mice as compared stage showed that projection neurons have fnally reached with controls. Furthermore, the capacity to induce LTP in the upper layer, similar to control. However, the morphology the CA1 was impaired in both withdrawn and alcoholized of neurons seems to be afected by prenatal alcohol exposure, mice. We conclude that early alcohol withdrawal and mode- especially at the level of apical dendrites. We thus plan to rate chronic alcoholization, have disrupting efects on spatial investigate more specifcally the terminal diferentiation and memory processes and synaptic plasticity in the CA1, leading dendritogenesis of projection neurons of alcohol-exposed to the compensatory use of striatum-dependent learning pups. We will also evaluate adult mice behavior and alcohol strategies. consumption in order to determine whether PAE has a long- term impact on adult behavior and drinking pattern. Lack of alpha5 nicotinic receptors increases alcohol self-administration at high dose and reverses the Characterization of the behavioral sensitization pattern of alcohol-induced neuronal activity in and the conditioned response induced by VTA and IPN long-term daily exposure to alcohol in DBA2/j 1 1 Léa Tochon ( Bordeaux, France) and Swiss mice Théo Van Ingelgom1 (1 Liège, Belgium) Impact of alcohol exposure on the development and maturation of the cerebral cortex 1 1 1 1 Studies on the locomotor sensitization induced by repeated Laura Van Hees , Sophie Laguesse , Laurent Nguyen ( Liège, ethanol administrations in mice use typically experimental Belgium) designs where duration of alcohol exposure is limited to

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 400 Congrès a maximum of 21 days. Consequently, little or nothing is (t=2.73; p=0.008), no diferences existing among cirrhotic known about sensitization induced by a more prolonged (17246±11021 pg/ml) and non-cirrhotic (21340±12442 ethanol exposure (exceeding 21 days). Terefore, the frst pg/ml). TGF-1 showed signifcant correlations with total aim of the present study is to characterize the behavioral cholesterol (r=0.28; p=0.017) and HDL-cholesterol (r=0.25; sensitization induced in mice by an extended period of daily p=0.042), and an inverse correlation with body mass index ethanol administrations (45 days). Te second aim of the pre- (BMI; =-0.37; p=0.004), IL-4 (=-0.31; p=0.009), INF- sent study is to test whether ethanol sensitization results in (=-0.28; p=0.001) or IL-6 a conditioned increase in locomotor activity when sensitized (=-0.38; p=0.001), but not with TNF- (=-0.02) or mice are confronted to a placebo test (saline injection) in the C-reactive protein (=-0.22, 0.06>p>0.05). By multivariate testing environment. Tis phenomenon, called conditioned analysis only BMI, IL-6 and HDL-cholesterol showed inde- response, has been well established with psychostimulants pendent relationships with TGF-1. No relationships were such as cocaine. However, the occurrence of an excitatory observed with ankle-brachial index or calcium in the aortic conditioned response after repeated exposure to a stimulant arch, hypertension, diabetes, left ventricular hypertrophy or dose of alcohol is still being debated. atrial fbrillation. Terefore TGF-1 levels are increased in For these purposes, Swiss and DBA/2J, the two most popu- alcoholics, but they are not related to vessel wall calcifcation lar mouse strains in the feld, received 45 consecutive daily or arterial stifness. ethanol administrations (respectively 2.5 and 2.0 g/kg) and their locomotor activity was daily recorded to test the deve- Brain-derived neurotrophic factor and handgrip lopment of ethanol sensitization. At the end of the procedure, strength among alcoholics a placebo test and a challenge test were conducted to assess Víctor E. Vera Delgado1, Lourdes Gonzalez Navarrete1, Lucía respectively the conditioned response and the inter-group Romero Acevedo1, Iván Ribot Hernández1, Candelaria Martín ethanol sensitization. González1, Elisa Espelosín-Ortega1, Melchor Rodríguez Gaspar1, Te results of the present study show that ethanol sensiti- Emilio González Reimers1 (1 Tenerife, Canary Islands, Spain) zation continues to develop beyond the usual duration of ethanol exposure used in the previous studies. Tus, ethanol sensitization reach maximal levels after about 25 injections Brain derived neurotrophic factor (BDNF) is involved in in DBA2/j mice and 40 injections in Swiss mice. However, neurogenesis and in the protection against oxidative damage it may be noted that the core phase of the development of and neuronal apoptosis. After exercise there is an increased ethanol sensitization occurred in both strains during the frst expression of this myokine, especially in skeletal muscle and 20 days. Remarkably, ethanol sensitization after such a long brain. Low BDNF levels have been described in neurode- daily ethanol treatment resulted in both an upward shift of generative diseases. Alcoholics show both muscle atrophy the magnitude of ethanol stimulant efects and a prolonga- and brain atrophy. Tus, this study was performed in order tion of these efects in time (up to 30 minutes). Finally, the to analyse the behavior of BDNF among alcoholics and its results of the placebo test clearly indicated an absence of association with brain atrophy and muscle mass and strength. conditioned response in both strain of mice. Serum BDNF values were prospectively determined to 82 male alcoholics (drinkers of 197±153 g ethanol/day during Arterial stiffness and TGF- among alcoholics 33±14 years), aged 58.62±11.21 years and 27 age-matched Víctor Vera-Delgado1, Lourdes González-Navarrete1, (54.52±7.78 years, Z=1.77; p=0.11) controls, and compared Selena Aguilera-García1, Melchor Rodríguez Gaspar1, with handgrip strength, with the presence of brain atrophy, Candelaria Martín-González1, Lucía Romero-Acevedo1, assessed by computed tomography (CT), and with the inten- Daniel Martínez-Martínez1, Emilio González-Reimers1 sity of alcoholism and liver function derangement. BDNF (1 Tenerife, Canary Islands, Spain) values were signifcantly lower among patients (median=6320, interquartile range=2404-12080 vs 19660(16228-27182 pg/ml, Z=6.36; p<0.001). Handgrip strength (signifcantly Transforming growth factor beta-1 (TGF- 1) is a pleiotropic reduced among patients) was correlated with BDNF values, cytokine. Its relationship with atherosclerosis is debatable, both in the whole population (=0.25; p<0.05), and, especial- protective or deleterious efects having been described. It has ly, in patients over 59 (median value) years (r=0.57, p<0.001). been reported that TGF- 1 is increased in alcoholics and BDNF was poorly related to liver function, but showed no heavily involved in liver fbrogenesis. However, its role on vas- relation at all with CT assessed brain atrophy. We conclude cular risk factors in these patients has not been analyzed. Tis that chronic alcoholics show decreased BDNF values that is the objective of this study. We included 79 heavy alcoholics are related to muscle function impairment rather than to age, and 34 controls. Calcium deposition in the aortic arch was brain atrophy, liver dysfunction, or the amount of ethanol assessed in the plain thorax X-ray flm. All the patients unde- consumed. rwent complete laboratory evaluation, including cholesterol fractions and serum levels of TGF-1, tumor necrosis factor Thiamine substitution in alcohol use disorder (TNF)- , interleukin (IL)-4, IL-6 and interferon- (IFNG). 1 2 1 Ankle-brachial index was recorded in 48 patients. Serum Nathalie Pruckner , Sandra Vyssoki , Benjamin Vyssoki 1 2 TGF-1 levels were signifcantly higher among patients ( Vienna, Austria, St. Pölten, Austria)

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 401 Congrès

Introduction: Patients with alcohol use disorder (AUD) indirect co-cultures by using the supernatant of SK hep cells frequently sufer from cognitive defcits ranging from mild treat Huh7 cells. Hepcidin, BMP6, BMP2, TFR1 and ferritin symptoms to most severe forms like Wernicke encephalo- were assessed by qRT-PCR and the Bmp6 concentration in pathy (WE). WE is caused by thiamine defciency and, if medium was determined by ELISA. left untreated, can progress to Korsakof syndrome, which Results: Treatment of SK hep cells with ferric iron led to constitutes severe anterograde amnesia, confabulation and signifcantly increased Bmp6 and hepcidin mRNA expression behavioral abnormalities. We conducted a review of the under hypoxia, whereas no efect on Huh7 cells was detected. current medical treatment guidelines for AUD in order to In direct co-cultures the mRNA expression levels have simi- identify recommendations for the use of thiamine. lar trends as found in SK hep and Huh7 single cultures. Of Methods: Tree diferent keyword combinations (“alcohol note, BMP6 expression in SK hep was extreme low and may treatment guideline”, “alcohol withdrawal guideline” and “al- not be sufcient to maintain adequate levels in the medium cohol treatment recommendation”) were entered in Pubmed to induce signaling in hepatocytes. and Scopus, additional guidelines were searched screening the Conclusion and outlook: SK hep cells are able to sense iron online sites of the respective agencies or societies. In total, 14 changes (iron supplementation or chelation) but no adequate guidelines were included. response of hepatocytes towards BMP6-induced signaling Results: Tiamine was mentioned in all but one of the re- could be observed. In the future, other EC lines (HUVECs), viewed publications. Specifcations on application modalities co-culture systems and 3D in vitro models will be explored to and indications varied considerably. While the majority of better understand the crosstalk on iron regulation. reviewed guidelines recommended parenteral thiamine only for patients at high risk for WE, some gave no information GDNF / Glia cell line derived neurotrophic factor – regarding the application form or dosage. A promising new treatment target in Alcohol use Conclusions: Substitution of parenteral thiamine in suspected disorder (AUD) WE is a well-established treatment regimen and high-dose Andreas Wippel1, Stephan Listabarth1, Andrea Gmeiner1, treatment with parenteral thiamine in several daily doses Daniel König1, Vid Velikic1, Benjamin Vyssoki1 (1 Vienna, Austria) should be considered a state-of-the-art procedure. Yet, hardly any evidence-based recommendations exist on a more gene- Background: According to the World Health organization ral use of thiamine as a preventative measure. Suggestions (WHO) alcohol accounted for 5,3% of all deaths worldwide according to medical guidelines vary widely. Further research in 2018. Alcohol is consumed globally but only a small part is of utmost importance to better defne and implement of consumers transit from social drinking habits to uncon- recommendations on use of thiamine in patients with alcohol trolled, compulsive drinking. Craving is a hallmark symptom use disorder. of alcohol dependence. Currently available anti-craving treatment options are limited in number, lack in efcacy and Hepatic iron overload in alcoholic liver disease: The face non-compliance in clinical use. Te dopaminergic system role of sinusoidal endothelial cells in iron sensing plays a key role in the reward circuit in dependence as well Shijin Wang1, Teresa Peccerella1, Sebastian Mueller1, as craving behavior. Vanessa Rausch1 (1 Heidelberg, Germany) Terefore, new treatment options should target neuronal reward pathways to normalize adaptations to chronic alcohol Background and aims: So far, hepatic iron overload in exposure and reduce craving. patients with alcoholic liver disease is poorly understood. Methods: Two diferent keyword combinations (“alcohol use Hepcidin, the master switch of systemic iron homeostasis disorder AND Gdnf ” “addiction AND Gdnf ”) were entered and is regulated by the BMP signaling pathway. Recent in PubMed. Relevant fndings are presented as a narrative data showed that liver sinusoidal endothelial cells (LSECs) review. express the highest amount of BMP6 among diferent hepa- Results: GDNF acts as a regulator of dopamine release and tic cell types and are able to regulate iron homeostasis in vivo. fring rates in the midbrain, especially in ventral tegmen- However, the exact mechanisms, how iron levels are sensed by tal area (VTA) and nucleus accumbens. Several studies in ECs and how BMP signaling is involved in the regulation of rodents suggest that GDNF is an alcohol responsive gene, systemic iron metabolism as well as which cells are involved which is upregulated in short term alcohol intake and down- is still not completely known. Te aim of this study is to regulated during withdrawal after excessive alcohol intake. establish an in vitro co-culture model to mimic the crosstalk Tese results were confrmed for humans in two independent between LSECs and hepatocytes for the investigation of the studies investigating GDNF serum levels and alcohol intake. exact role of LSECs in regulating iron metabolism. Furthermore, elevated GDNF produced suppression of alco- Methods: Huh7 cells (hepatocytes) and SK hep cells (endo- hol-drinking behaviours in rats and reduced GDNF facilita- thelial cells) were cultured alone and treated with increasing ted the escalation of alcohol drinking. concentrations of ferric iron and with two iron chelators (des- Discussion: Te escalation from moderate to excessive drin- feral and SIH) under normoxic (21% O2) as well as hypoxic king could be a result of a breakdown of endogenous GDNF conditions (1% O2) for 24 hours. Next, direct co-cultures with systems, therefore, GDNF could be a marker for AUD and SK hep and Huh7 cells were established by inserts as well as may serve as a treatment target to reduce craving.

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 402 Congrès

Is (poly-) substance use associated with impaired tobacco use and response inhibition (in the SST) difered inhibitory control? A mega-analysis controlling for between cannabis users and non-users, with a negative asso- confounders ciation between tobacco use and inhibition in the cannabis Yang Liu 1, Wery P.M. van den Wildenberg1, Ysanne de Graaf1, non-users. In addition, participants’ age, education level, and Susan L. Ames2, Alexander Baldacchino3, Ragnhild Bø4, Fernando some task characteristics infuenced inhibition outcomes. Cadaveira5, Salvatore Campanella6, Paul Christiansen7, Eric D. Overall, we found limited support for impaired inhibition Claus8, Lorenza S. Colzato9, Francesca M. Filbey10, John J. Foxe11, among substance users when controlling for demographics Hugh Garavan12, Christian S. Hendershot13, Robert Hester14, and task-characteristics. Jennifer M. Jester15, Hollis C. Karoly16, Anja Kräplin17, Fanny Role of NOX1 on hepcidin signaling in the Kreusch18, Nils Inge Landrø4, Marianne Littel19, Sabine Steins- crosstalk between macrophages and hepatocytes Loeber 20, Edythe D. London21, Eduardo López-Caneda22, Dan I. Linna Yu1, Teresa Peccerella1, Sebastian Mueller1, Lubman14, Maartje Luijten23, Cecile A. Marczinski24, Jane Metrik25, Vanessa Rausch1 (1 Heidelberg, Germany) Catharine Montgomery26, Harilaos Papachristou27, Su Mi Park28, 29 7 30 Andres L. Paz , Géraldine Petit , James J. Prisciandaro , Boris B. Background and aims: Liver-secreted hepcidin is the syste- Quednow31, Lara A. Ray21, Carl A. Roberts7, Gloria M.P. Roberts32, mic masterswitch of iron homeostasis and its dysregulation Michiel B. de Ruiter1, Claudia I. Rupp33, Vaughn R. Steele8, leads to iron accumulation in most of chronic liver diseases. 34 14 35 Delin Sun , Michael Takagi , Susan F. Tapert , Ruth J. van Hepcidin is regulated by iron, infammation or H2O2, but 1 36 31 Holst , Antonio Verdejo-Garcia , Matthias Vonmoos , Marcin the role of NOX1 and its products ROS/H2O2 in monocyte- Wojnar37, Yuanwei Yao38, Murat Yücel14, Martin Zack13, Robert A. derived macrophages on hepcidin regulation under (patho) Zucker15, Hilde M. Huizenga1, Reinout W. Wiers1 (1 Amsterdam, physiological conditions is poorly understood. We here inves- The Netherlands, 2 Claremont, USA, 3 St Andrews, Scotland, tigate the role of NOX1 on regulating hepcidin and cytokines United Kingdom, 4 Oslo, Norway, 5 Santiago de Compostela, in infammatory macrophages and subsequent efects on Spain, 6 Brussels, Belgium, 7 Nicosia, Cyprus, 8 Albuquerque, New hepatocytes mimicking (patho)physiological conditions (cell Mexico, 9 Leiden, the Netherlands, 10 Dallas, USA, 11 Rochester, ratios, oxygen levels and infammation). USA, 12 Burlington, USA, 13 Toronto, Canada, 14 Melbourne, Methods: THP-1 monocytes were diferentiated into macro- Australia, 15 Ann Arbor, USA, 16 Boulder, USA, 17 Dresden, phages and co-cultured with Huh7 cells at physiological cell Germany, 18 Liège, Belgium, 19 Rotterdam, The Netherlands, ratio (4:1) and treated with diferent LPS concentrations 20 Bamberg, Germany, 21 Los Angeles, USA, 22 Braga, Portugal, (10 ng/ml and 100ng/ml) under normoxia (21% O2) or 23 Nijmegen, The Netherlands, 24 Highland Heights, USA, hypoxia (1% O2). Te exposure of Huh7 cells to macro- phage-conditioned medium with LPS was also investigated. 25 Providence, USA, 26 Liverpool, United Kingdom, 27 Maastricht, Hepcidin, IL-1 IL-6, C/EBP and SMAD6 mRNA levels The Netherlands, 28 Seoul, Republic of Korea, 29 Boca Raton, USA, were assessed by qRT-PCR and the expression of NOX1, p- 30 Charleston, USA, 31 Zürich, Switzerland, 32 Sydney, Australia, 33 34 35 STAT3, STAT3 and p-SMAD1/5/8 proteins was analyzed Innsbruck, Austria, Durham, USA, San Diego, USA, by western blot. 36 37 38 Clayton, Australia, Warsaw, Poland, Beijing, China) Results: LPS signifcantly increased NOX1, p-STAT3, IL- 1and IL-6 levels in THP-1 macrophages, but decreased Many studies have reported that heavy substance use is asso- STAT3 expression in a concentration-dependent manner ciated with impaired response inhibition. Studies typically under 21% and 1% O2. Interestingly, 10ng/ml LPS increased focused on associations with a single substance, while poly- the expression of hepcidin whereas 100ng/ml LPS decreased substance use is common. Further, most studies compared the expression of hepcidin under 21% O2. In contrast, both heavy users with light/non-users, though substance use occurs LPS concentrations decreased the expression of hepcidin un- along a continuum. Te current mega-analysis accounted for der low oxygen conditions (1% O2) in THP-1 macrophages. these issues by aggregating individual data from 43 studies In addition, LPS decreased SMAD6, p-SMAD1/5/8 and (3610 adult participants) that used the Go/No-Go (GNG) CEBP in THP-1 macrophages under normoxia (21% O2). or Stop-signal task (SST) to assess inhibition among mostly Conclusion: Our findings underscore a possible role of “recreational” substance users (i.e., the rate of substance use NOX1 and subsequent ROS/H2O2 concentrations on hep- disorders was low). Main and interaction efects of substance cidin regulation and induction of cytokine production in use, demographics, and task-characteristics were entered in infammatory macrophages involving the STAT3 signaling a linear mixed model. Contrary to many studies and reviews pathway. In the future, we aim at studying in detail hepcidin in the feld, we found that only lifetime cannabis use was signaling by using WT and truncated hepcidin promoter associated with impaired response inhibition in the SST. An constructs and siRNA-mediated knockdown of TLR4, interaction efect was also observed: the relationship between NOX1, STAT3 or C/EBP.

Alcoologie et Addictologie. 2019 ; 41 (4) : 368-403 403

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