DOCSLIB.ORG

Mexican Carriers of the HNF1A P.E508K Variant Do Not Experience

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Mexican Carriers of the HNF1A P.E508K Variant Do Not Experience 1726 Diabetes Care Volume 41, August 2018 Mexican Carriers of the HNF1A Alexandro J. Martagon,´ 1,2 Omar Yaxmehen Bello-Chavolla,1,3 p.E508K Variant Do Not Olimpia Arellano-Campos,1 Paloma Almeda-Valdes,´ 1,4 Experience an Enhanced Response Geoffrey A. Walford,5,6,7 Ivette Cruz-Bautista,1,4 to Sulfonylureas Donaj´ı V.Gomez-Velasco,´ 1 RoopaMehta,1,4 Liliana Munoz-Hern~ andez,´ 1 1 Diabetes Care 2018;41:1726–1731 | https://doi.org/10.2337/dc18-0384 Magdalena Sevilla-Gonzalez,´ Tannia L. Viveros-Ruiz,1 Mar´ıa Luisa Ordonez-S~ anchez,´ 8 Rosario Rodr´ıguez-Guillen,8 Jose C. Florez,5,6,7 Mar´ıa Teresa Tusie-Luna,´ 8 and Carlos A. Aguilar-Salinas,1,2,4 on behalf of the Slim Initiative in Genomic Medicine for the Americas (SIGMA) Type 2 Diabetes OBJECTIVE Consortium* To assess whether an ethnic-specific variant (p.E508K) in the maturity-onset dia- betes of the young (MODY) gene hepatocyte nuclear factor-1a (HNF1A) found in 1Unidad de Investigacion´ de Enfermedades Mexicans is associated with higher sensitivity to sulfonylureas, as documented Metabolicas,´ Instituto Nacional de Ciencias in patients with MODY3. Medicas´ y Nutricion,´ Ciudad de Mexico,´ Mexico´ 2Tecnologico´ de Monterrey, Escuela de Medicina RESEARCH DESIGN AND METHODS y Ciencias de la Salud, Monterrey, Nuevo Leon,´ Mexico´ We recruited 96 participants (46 variant carriers and 50 age- and sex-matched 3 – Plan de Estudios Combinados en Medicina, noncarriers). Response to glipizide (one 2.5 5.0-mg dose), metformin (four 500-mg Facultad de Medicina, Universidad Nacional doses), and an oral glucose challenge was evaluated using a previously validated Autonoma´ de Mexico,´ Ciudad de Mexico,´ Mexico´ protocol. Glucose and insulin levels and their areas under the curve (AUCs) were 4Departamento de Endocrinolog´ıa y Metabo- compared between groups. lismo, Instituto Nacional de Ciencias Medicas´ y Nutricion´ Salvador Zubiran,´ Ciudad de Mexico,´ RESULTS Mexico´ 5Center for Genomic Medicine and Diabetes Unit, Carriers of the p.E508K variant had a lower maximum insulin peak during the Massachusetts General Hospital, Boston, MA glipizide challenge as compared with noncarriers with diabetes (P < 0.05). Also, 6Programs in Metabolism and Medical and carriers had a lower insulin response after the oral glucose challenge. Following an Population Genetics, Broad Institute of MIT and Harvard, Cambridge, MA oral glucose tolerance test in the presence of metformin, carriers of the p.E508K 7Harvard Medical School, Boston, MA variant with diabetes had a lower maximum insulin peak and total and incre- 8Unidad de Biolog´ıa Molecular y Medicina Geno-´ mental insulin AUC value as compared with noncarriers with diabetes (P < 0.05). A mica, Instituto Nacional de Ciencias Medicas´ y similar but nonsignificant trend was seen in participants without type 2 diabetes. Nutricion´ Salvador Zubiran,´ Instituto de Inves- PATHOPHYSIOLOGY/COMPLICATIONS tigaciones Biomedicas,´ Universidad Nacional CONCLUSIONS Autonoma´ de Mexico,´ Ciudad de Mexico,´ Mexico´ HNF1A Corresponding author: Carlos A. Aguilar-Salinas, Carriers of variant p.E508K in have a reduced insulin response rather than [email protected]. the increased sensitivity to sulfonylureas seen in patients with MODY3. Received 21 February 2018 and accepted 1 May 2018. Type 2 diabetes is the leading cause of death and a major burden for public health of This article contains Supplementary Data online at http://care.diabetesjournals.org/lookup/suppl/ Mexican and Latino populations (1). Ethnic-specific genetic variants have been de- doi:10.2337/dc18-0384/-/DC1. scribed in populations with a Native American heritage (such as Mexicans). The Slim *A complete list of the members of the Slim Initiative in Genomic Medicine for the Americas (SIGMA) Type 2 Diabetes Consor- Initiative in Genomic Medicine for the Americas tium has reported associations with a common haplotype in SLC16A11 (2) through a Type 2 Diabetes Consortium can be found in the genome-wide association study in ;9,000 participants and a rare missense variant Supplementary Data online. (c.1522G . A [p.E508K]) in the gene encoding the hepatocyte nuclear factor-1a © 2018 by the American Diabetes Association. (HNF1A) through whole-exome sequencing in ;4,000 participants (3), both of which Readers may use this article as long as the work – is properly cited, the use is educational and not are rare or absent in non Native American populations. The Mexican population for profit, and the work is not altered. More infor- results from a recent admixture of European and Native American populations with mation is available at http://www.diabetesjournals similar proportions and a relatively low African ancestry (,5%). This admixture is .org/content/license. care.diabetesjournals.org Martagon´ and Associates 1727 known as the mestizo population. There- RESEARCH DESIGN AND METHODS or women at risk of becoming pregnant; if fore, exome-sequencing in Mexican mes- The study was supported by the SIGMA they had age of onset of diabetes before tizos has resulted in the identification of Type 2 Diabetes Consortium and per- 25 years of age, known history of liver or genetic variants mainly derived from our formed in accordance with the SUGAR- kidney disease, allergy to sulfonamides, Native American heritage. In particular, the MGH protocol (10). The study protocol history of porphyria, impaired renal func- HNF1A variant, located in exon 8, causes a was approved by the Ethics Commit- tion (estimated glomerular filtration rate 2 partial defect in the function of the tran- tee of the Instituto Nacional de Ciencias ,60 mL/min/1.73 m ), established coro- scription factor (3), which is expressed in Medicas´ y Nutricion´ Salvador Zubiran.´ nary artery disease, or history of bariatric liver and pancreas. Though the p.E508K surgery, seizures, or stroke; if they were variant is associated with a fivefold in- Subjects and Setting taking medications that could affect creased risk for developing type 2 diabe- Mexican mestizos (defined as Mexican glycemic parameters; or if they were tes, patients with the variant were clinically individuals whose father, mother, and planning radiologic or angiographic stud- undistinguishable in terms of age of onset, grandfathers were born in Mexico and ies requiring contrast within 1 week of adiposity, and glycemia from type 2 diabe- who do not belong to any other ethnic completion of the study. All participants tes case subjects (3). The variant is present group [i.e., Jewish, Japanese, etc.]) aged read and signed an informed consent in 0.2% of the Mexican participants with $18 years, males or nonpregnant fe- document. type 2 diabetes in the SIGMA exome- males, healthy control subjects (n =33), sequencing analyses (3). Loss-of-function and patients with type 2 diabetes (n =16) Interventions mutations in HNF1A cause maturity-onset were recruited at the diabetes outpatient The study consisted of two visits (Fig. 1) diabetes of the young type 3 (MODY3) clinic of the Instituto Nacional de Ciencias (10). Prior to visit 1, participants with (4,5). MODY3 is characterized by an early Medicas´ y Nutricion´ Salvador Zubiran´ in type 2 diabetes taking an oral antidia- onset of the disease, normal body weight, Mexico City, a center attending indivi- betic agent underwent a 7-day washout glycosuria, and high risk for microvascular duals from the general population. An period. Blood samples were obtained complications (6–8). In addition, patients additional recruitment of carriers and non- between 8:00 A.M. and 9:00 A.M. in the with MODY3 have high sensitivity to sul- carriers of the p.E508K HNF1A variant morning after an 8–12-h overnight fast. fonylureas; this feature underlies the cur- was done among users of the diabetes A complete medical and family history rent indication for sulfonylurea therapy in outpatient clinic of the Instituto Nacional as well as anthropometric measurements these patients, over metformin or insulin de Ciencias Medicas´ y Nutricion´ or were obtained. Participants were weighed (8,9), and represents one successful exam- among the first-degree relatives of the on calibrated scales, and height was de- ple of precision medicine in diabetes. E508K variant carriers. Genotyping of the termined with a floor scale stadiometer; Because p.E508K causes partial loss p.E508K variant (rs483353044) was per- BMI was calculated as weight in kilograms of function in HNF1A, we hypothesized formed using a TaqMan probe. For the divided by the square of height in me- that carriers of this variant might also ex- assay, we included control subjects pre- ters. Participants with a fasting glucose hibit heightened sensitivity to sulfony- viously sequenced and selected carry- #80 mg/dL were not eligible to receive lureas. The aim of our study was to advance ing the three possible genotypes for this the sulfonylurea challenge for safety rea- precision medicine by evaluating the insul- single nucleotide polymorphism. Case sub- sons; those with fasting glucose 80–99 in and glycemic responses to glipizide and jects with type 2 diabetes were eligible if mg/dL received 2.5 mg of glipizide, metformin in p.E508K carriers and age- they were being treated with no more and those with a fasting glucose $100 and sex-matched noncarriers. We fol- than two oral antidiabetic agents and mg/dL received 5 mg. After glipizide ad- lowed the approach used in the Study hadanHbA1c #7.5% (58 mmol/mol). Non- ministration, blood samples were col- to Understand the Genetics of the Acute carriers were matched by age (65years), lected at 30, 60, 90, 120, 180, and Response to Metformin and Glipizide sex, BMI (62kg/m2), and status of diabetes. 240 min for glucose and insulin mea- in Humans (SUGAR-MGH), a standard- Volunteers were excluded from par- surements. After the 240-min period, ized pharmacogenetic protocol (10,11). ticipation if they were pregnant, nursing, breakfast was given, and patients were Figure 1—Outline of the study period and intervention.
Load more

Recommended publications
  • Mediator of DNA Damage Checkpoint 1 (MDC1) Is a Novel Estrogen Receptor Co-Regulator in Invasive 6 Lobular Carcinoma of the Breast 7 8 Evelyn K
    bioRxiv preprint doi: https://doi.org/10.1101/2020.12.16.423142; this version posted December 16, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC 4.0 International license. 1 Running Title: MDC1 co-regulates ER in ILC 2 3 Research article 4 5 Mediator of DNA damage checkpoint 1 (MDC1) is a novel estrogen receptor co-regulator in invasive 6 lobular carcinoma of the breast 7 8 Evelyn K. Bordeaux1+, Joseph L. Sottnik1+, Sanjana Mehrotra1, Sarah E. Ferrara2, Andrew E. Goodspeed2,3, James 9 C. Costello2,3, Matthew J. Sikora1 10 11 +EKB and JLS contributed equally to this project. 12 13 Affiliations 14 1Dept. of Pathology, University of Colorado Anschutz Medical Campus 15 2Biostatistics and Bioinformatics Shared Resource, University of Colorado Comprehensive Cancer Center 16 3Dept. of Pharmacology, University of Colorado Anschutz Medical Campus 17 18 Corresponding author 19 Matthew J. Sikora, PhD.; Mail Stop 8104, Research Complex 1 South, Room 5117, 12801 E. 17th Ave.; Aurora, 20 CO 80045. Tel: (303)724-4301; Fax: (303)724-3712; email: [email protected]. Twitter: 21 @mjsikora 22 23 Authors' contributions 24 MJS conceived of the project. MJS, EKB, and JLS designed and performed experiments. JLS developed models 25 for the project. EKB, JLS, SM, and AEG contributed to data analysis and interpretation. SEF, AEG, and JCC 26 developed and performed informatics analyses. MJS wrote the draft manuscript; all authors read and revised the 27 manuscript and have read and approved of this version of the manuscript.
    [Show full text]
  • The Roles of Long Noncoding Rnas HNF1-AS1 and HNF4-AS1 in Drug
    non-coding RNA Review The Roles of Long Noncoding RNAs HNF1α-AS1 and HNF4α-AS1 in Drug Metabolism and Human Diseases Liming Chen 1, Yifan Bao 1, Suzhen Jiang 1,2 and Xiao-bo Zhong 1,* 1 Department of Pharmaceutical Sciences, School of Pharmacy, University of Connecticut, Storrs, CT 06269, USA; [email protected] (L.C.); [email protected] (Y.B.); [email protected] (S.J.) 2 School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 51006, China * Correspondence: [email protected]; Tel.: +01-860-486-3697 Received: 25 May 2020; Accepted: 22 June 2020; Published: 24 June 2020 Abstract: Long noncoding RNAs (lncRNAs) are RNAs with a length of over 200 nucleotides that do not have protein-coding abilities. Recent studies suggest that lncRNAs are highly involved in physiological functions and diseases. lncRNAs HNF1α-AS1 and HNF4α-AS1 are transcripts of lncRNA genes HNF1α-AS1 and HNF4α-AS1, which are antisense lncRNA genes located in the neighborhood regions of the transcription factor (TF) genes HNF1α and HNF4α, respectively. HNF1α-AS1 and HNF4α-AS1 have been reported to be involved in several important functions in human physiological activities and diseases. In the liver, HNF1α-AS1 and HNF4α-AS1 regulate the expression and function of several drug-metabolizing cytochrome P450 (P450) enzymes, which also further impact P450-mediated drug metabolism and drug toxicity. In addition, HNF1α-AS1 and HNF4α-AS1 also play important roles in the tumorigenesis, progression, invasion, and treatment outcome of several cancers. Through interacting with different molecules, including miRNAs and proteins, HNF1α-AS1 and HNF4α-AS1 can regulate their target genes in several different mechanisms including miRNA sponge, decoy, or scaffold.
    [Show full text]
  • Homozygous Hypomorphic HNF1A Alleles Are a Novel Cause of Young-Onset Diabetes and Result in Sulphonylurea-Sensitive Diabetes
    Diabetes Care 1 Shivani Misra,1 Neelam Hassanali,2 Homozygous Hypomorphic Amanda J. Bennett,2 Agata Juszczak,2 Richard Caswell,3 Kevin Colclough,3 HNF1A Alleles Are a Novel Cause Jonathan Valabhji,4 Sian Ellard,3 of Young-Onset Diabetes and Nicholas S. Oliver,1,4 and Anna L. Gloyn2,5,6 Result in Sulphonylurea-Sensitive Diabetes https://doi.org/10.2337/dc19-1843 OBJECTIVE Heterozygous loss-of-function mutations in HNF1A cause maturity-onset diabetes of the young (MODY). Affected individuals can be treated with low-dose sulpho- nylureas. Individuals with homozygous HNF1A mutations causing MODY have not been reported. RESEARCH DESIGN AND METHODS We phenotyped a kindred with young-onset diabetes and performed molecular genetic testing, a mixed meal tolerance test, a sulphonylurea challenge, and in vitro assays to assess variant protein function. RESULTS A homozygous HNF1A variant (p.A251T) was identified in three insulin-treated 1 family members diagnosed with diabetes before 20 years of age. Those with the Diabetes, Endocrinology and Metabolism, Im- NOVEL COMMUNICATIONS IN DIABETES homozygous variant had low hs-CRP levels (0.2–0.8 mg/L), and those tested dem- perial College London, London, U.K. 2Oxford Centre for Diabetes, Endocrinology and onstrated sensitivity to sulphonylurea given at a low dose, completely transitioning off Metabolism, University of Oxford, Oxford, U.K. insulin. In silico modeling predicted a variant of unknown significance; however, in vitro 3Institute of Biomedical and Clinical Science, Uni- studies supported a modest reduction in transactivation potential (79% of that for the versity of Exeter Medical School, Exeter, U.K.
    [Show full text]
  • Transcriptome Analysis Uncovers the Diagnostic Value of Mir-192-5P/HNF1A-AS1/VIL1 Panel in Cervical Adenocarcinoma
    www.nature.com/scientificreports OPEN Transcriptome analysis uncovers the diagnostic value of miR‑192‑5p/ HNF1A‑AS1/VIL1 panel in cervical adenocarcinoma Junfen Xu1*, Jian Zou1, Luyao Wu1 & Weiguo Lu1,2* Despite the fact that the incidence of cervical squamous cell carcinoma has decreased, there is an increase in the incidence of cervical adenocarcinoma. However, our knowledge on cervical adenocarcinoma is largely unclear. Transcriptome sequencing was conducted to compare 4 cervical adenocarcinoma tissue samples with 4 normal cervical tissue samples. mRNA, lncRNA, and miRNA signatures were identifed to discriminate cervical adenocarcinoma from normal cervix. The expression of VIL1, HNF1A‑AS1, MIR194‑2HG, SSTR5‑AS1, miR‑192‑5p, and miR‑194‑5p in adenocarcinoma were statistically signifcantly higher than that in normal control samples. The Receiver Operating Characteristic (ROC) curve analysis indicated that combination of miR‑192‑5p, HNF1A‑AS1, and VIL1 yielded a better performance (AUC = 0.911) than any single molecule -and could serve as potential biomarkers for cervical adenocarcinoma. Of note, the combination model also gave better performance than TCT test for cervical adenocarcinoma diagnosis. However, there was no correlation between miR‑192‑5p or HNF1A‑AS1 and HPV16/18 E6 or E7. VIL1 was weakly correlated with HPV18 E7 expression. In summary, our study has identifed miR‑192‑5p/HNF1A‑AS1/VIL1 panel that accurately discriminates adenocarcinoma from normal cervix. Detection of this panel may provide considerable clinical value in the diagnosis of cervical adenocarcinoma. Abbreviations SCC Cervical squamous cell carcinoma NcRNA Non-coding RNA MiRNA MicroRNA LncRNA Long non-coding RNA ROC Receiver operating characteristic AUC​ Area under the ROC curve RT-qPCR Quantitative reverse-transcriptase PCR HNF1A-AS1 LncRNA HNF1A antisense RNA 1 VIL1 Villin 1 Cervical cancer ranks fourth for both cancer incidence and mortality in women worldwide 1.
    [Show full text]
  • Hepatocyte Nuclear Factor 1Αsuppresses Steatosis- Associated
    The Journal of Clinical Investigation RESEARCH ARTICLE Hepatocyte nuclear factor 1α suppresses steatosis- associated liver cancer by inhibiting PPARγ transcription Cecilia Patitucci,1,2,3 Gabrielle Couchy,4 Alessia Bagattin,3,5 Tatiana Cañeque,6,7,8 Aurélien de Reyniès,9 Jean-Yves Scoazec,10 Raphaël Rodriguez,6,7,8 Marco Pontoglio,3,5 Jessica Zucman-Rossi,3,4,11,12,13 Mario Pende,1,2,3 and Ganna Panasyuk1,2,3 1Institut Necker-Enfants Malades, 2INSERM U1151/CNRS Unité Mixte de Recherche (UMR) 8253, Paris, France. 3Université Paris Descartes, Sorbonne Paris Cité, Paris, France. 4INSERM, UMR 1162, Functional Genomics of Solid Tumors, Equipe Labellisée Ligue Contre le Cancer, Paris, France. 5INSERM U1016/CNRS UMR 8104, Institut Cochin, Paris, France. 6Institut Curie, PSL Research University, Organic Synthesis and Cell Biology Group, Paris, France. 7CNRS UMR 3666, Paris, France. 8INSERM U1143, Paris, France. 9Ligue Nationale Contre le Cancer, Paris, France. 10INSERM, UMR 865, Faculté Laennec, Lyon, France. 11Hopital Europeen Georges Pompidou, Paris, France. 12University of Paris Diderot, Sorbonne Paris Cité, University Institute of Hematology, Paris, France. 13University of Paris, Sorbonne Paris Cité, Saint-Denis, France. Worldwide epidemics of metabolic diseases, including liver steatosis, are associated with an increased frequency of malignancies, showing the highest positive correlation for liver cancer. The heterogeneity of liver cancer represents a clinical challenge. In liver, the transcription factor PPARγ promotes metabolic adaptations of lipogenesis and aerobic glycolysis under the control of Akt2 activity, but the role of PPARγ in liver tumorigenesis is unknown. Here we have combined preclinical mouse models of liver cancer and genetic studies of a human liver biopsy atlas with the aim of identifying putative therapeutic targets in the context of liver steatosis and cancer.
    [Show full text]
  • Proximal Tubule Translational Profiling During Kidney Fibrosis Reveals Pro- Inflammatory and Lncrna Expression Patterns with Sexual Dimorphism
    BASIC RESEARCH www.jasn.org Proximal Tubule Translational Profiling during Kidney Fibrosis Reveals Proinflammatory and Long Noncoding RNA Expression Patterns with Sexual Dimorphism Haojia Wu,1,2 Chun-Fu Lai,1,2,3 Monica Chang-Panesso,1,2 and Benjamin D. Humphreys 1,2,4 1Division of Nephrology, Departments of 2Medicine and 4Developmental Biology, Washington University in St. Louis School of Medicine, St. Louis, Missouri; and 3Renal Division, Department of Internal Medicine, National Taiwan University Hospital, Taipai, Taiwan ABSTRACT Background Proximal tubule injury can initiate CKD, with progression rates that are approximately 50% faster in males versus females. The precise transcriptional changes in this nephron segment during fibrosis and potential differences between sexes remain undefined. Methods We generated mice with proximal tubule–specific expression of an L10a ribosomal subunit pro- tein fused with enhanced green fluorescent protein. We performed unilateral ureteral obstruction surgery on four male and three female mice to induce inflammation and fibrosis, collected proximal tubule–specific and bulk cortex mRNA at day 5 or 10, and sequenced samples to a depth of 30 million reads. We applied computational methods to identify sex-biased and shared molecular responses to fibrotic injury, including up- and downregulated long noncoding RNAs (lncRNAs) and transcriptional regulators, and used in situ hybridization to validate critical genes and pathways. Results We identified .17,000 genes in each proximal tubule group, including 145 G-protein–coupled receptors. More than 700 transcripts were differentially expressed in the proximal tubule of males versus females. The .4000 genes displaying altered expression during fibrosis were enriched for proinflam- matory and profibrotic pathways.
    [Show full text]
  • ORIGINAL ARTICLE Correlation Between Density of CD8 + T Cell
    Author Manuscript Published OnlineFirst on June 9, 2015; DOI: 10.1158/0008-5472.CAN-14-3051 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. ORIGINAL ARTICLE Correlation between density of CD8+ T cell infiltrates in microsatellite unstable colorectal cancers and frameshift mutations: a rationale for personalized immunotherapy Running title: CD8+ cell density correlates with mutations in MSI-CRCs Pauline Maby1; David Tougeron1,2,3; Mohamad Hamieh1; Bernhard Mlecnik4,5,6; Hafid Kora1; Gabriela Bindea4,5,6; Helen K Angell4,5,6,7; Tessa Fredriksen4,5,6; Nicolas Elie8; Emilie Fauquembergue1; Aurélie Drouet1; Jérôme Leprince9; Jacques Benichou10; Jacques Mauillon11,12; Florence Le Pessot13; Richard Sesboüé1; Thierry Frébourg1,11; Jérôme Galon4,5,6; Jean-Baptiste Latouche1,11 1Inserm U1079, Institute for Research and Innovation in Biomedecine (IRIB), Rouen, France; 2Gastroenterology Department, Poitiers University Hospital, Poitiers, France; 3Laboratoire Inflammation Tissus Epithéliaux et Cytokines, EA 4331, Poitiers University, Poitiers, France; 4 Inserm U1138, Laboratory of Integrative Cancer Immunology, Paris, France; 5Université Paris Descartes, Paris, France; 6Cordeliers Research Centre, Université Pierre et Marie Curie, Paris 6, Paris, France; 7AstraZeneca Pharmaceuticals, Alderley Park, Cheshire, UK 8Imaging Core Facility, CMABIO, Caen University Hospital, Caen, France; 9Inserm U982, Institute for Research and Innovation in Biomedecine (IRIB), Rouen University, France 1 Downloaded from
    [Show full text]
  • Impact of Hypoxia on Hepatitis B Virus Replication
    i IMPACT OF HYPOXIA ON HEPATITIS B VIRUS REPLICATION NICHOLAS ROSS BAKER FRAMPTON A thesis submitted to the University of Birmingham for the degree of Doctor of Philosophy Institute of Immunology and Immunotherapy College of Medical and Dental Sciences University of Birmingham September 2017 ii University of Birmingham Research Archive e-theses repository This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder. ABSTRACT Hepatitis B virus (HBV) is one of the world’s unconquered diseases, with 370 million chronically infected globally. HBV replicates in hepatocytes within the liver that exist under a range of oxygen tensions from 11% in the peri-portal area to 3% in the peri- central lobules. HBV transgenic mice show a zonal pattern of viral antigen with expression in the peri-central areas supporting a hypothesis that low oxygen regulates HBV replication. We investigated this hypothesis using a recently developed in vitro model system that supports HBV replication. We demonstrated that low oxygen significantly increases covalently closed circular viral DNA (cccDNA), viral promoter activity and pre-genomic RNA (pgRNA) levels, consistent with low oxygen boosting viral transcription. Hypoxia inducible factors (HIFs) regulate cellular responses to low oxygen and we investigated a role for HIF-1α or HIF-2α on viral transcription.
    [Show full text]
  • BMAL1 and Modulates Tissue-Specific Circadian Networks
    Nuclear receptor HNF4A transrepresses CLOCK: BMAL1 and modulates tissue-specific circadian networks Meng Qua, Tomas Duffyb, Tsuyoshi Hirotac, and Steve A. Kaya,1 aKeck School of Medicine, University of Southern California, Los Angeles, CA 90089; bDepartment of Molecular Medicine, The Scripps Research Institute, La Jolla, CA 92037; and cInstitute of Transformative Bio-Molecules, Nagoya University, 464-8602 Nagoya, Japan Contributed by Steve A. Kay, November 6, 2018 (sent for review September 24, 2018; reviewed by Carla B. Green and John B. Hogenesch) Either expression level or transcriptional activity of various nuclear NRs canonically function as ligand-activated transcription receptors (NRs) have been demonstrated to be under circadian factors that regulate the expression of their target genes to control. With a few exceptions, little is known about the roles of affect physiological pathways (19). The importance of NRs in NRs as direct regulators of the circadian circuitry. Here we show maintaining optimal physiological homeostasis is illustrated in that the nuclear receptor HNF4A strongly transrepresses the their identification as potential targets for therapeutic drug transcriptional activity of the CLOCK:BMAL1 heterodimer. We development to combat a diverse array of diseases, including define a central role for HNF4A in maintaining cell-autonomous reproductive disorders, inflammation, cancer, diabetes, car- circadian oscillations in a tissue-specific manner in liver and colon diovascular disease, and obesity (20). Various NRs have been cells. Not only transcript level but also genome-wide chromosome implicated as targets of the circadian clock, which may con- binding of HNF4A is rhythmically regulated in the mouse liver. tribute to the circadian regulation of nutrient and energy me- ChIP-seq analyses revealed cooccupancy of HNF4A and CLOCK: tabolism.
    [Show full text]
  • Spectrum of HNF1A Somatic Mutations in Hepatocellular
    ORIGINAL ARTICLE Spectrum of HNF1A Somatic Mutations in Hepatocellular Adenoma Differs From That in Patients With MODY3 and Suggests Genotoxic Damage Emmanuelle Jeannot,1,2 Lucille Mellottee,1 Paulette Bioulac-Sage,3 Charles Balabaud,3 Jean-Yves Scoazec,4 Jeanne Tran Van Nhieu,5 Yannick Bacq,6 Sophie Michalak,7 David Buob,8 Groupe d’e´tude Ge´ne´tique des Tumeurs He´patiques (INSERM Network), Pierre Laurent-Puig,9 Ivan Rusyn,2 and Jessica Zucman-Rossi1 OBJECTIVE—Maturity onset diabetes of the young type 3 (MODY3) is a consequence of heterozygous germline mutation in HNF1A. A subtype of hepatocellular adenoma (HCA) is also epatocellular adenoma (HCA) is a rare, benign, caused by biallelic somatic HNF1A mutations (H-HCA), and rare liver tumor frequently associated with oral con- HCA may be related to MODY3. To better understand a relation- traception (1,2). HCA usually manifests as a ship between the development of MODY3 and HCA, we com- single tumor, but in some cases, several adeno- pared both germline and somatic spectra of HNF1A mutations. H mas are detected in the same patient; when Ͼ10 nodules RESEARCH DESIGN AND METHODS—We compared 151 are identified in the liver it is called liver adenomatosis (3). somatic HNF1A mutations in HCA with 364 germline mutations Recently, by the analysis of a large series of patients with described in MODY3. We searched for genotoxic and oxidative HCAs, we established a new molecular classification of stress features in HCA and surrounding liver tissue. these tumors. Adenomas were classified according to the RESULTS—A spectrum of HNF1A somatic mutations signifi- genotype of the tumors, such as the finding of mutations in cantly differed from the germline changes in MODY3.
    [Show full text]
  • Integrated Computational Approach to the Analysis of RNA-Seq Data Reveals New Transcriptional Regulators of Psoriasis
    OPEN Experimental & Molecular Medicine (2016) 48, e268; doi:10.1038/emm.2016.97 & 2016 KSBMB. All rights reserved 2092-6413/16 www.nature.com/emm ORIGINAL ARTICLE Integrated computational approach to the analysis of RNA-seq data reveals new transcriptional regulators of psoriasis Alena Zolotarenko1, Evgeny Chekalin1, Alexandre Mesentsev1, Ludmila Kiseleva2, Elena Gribanova2, Rohini Mehta3, Ancha Baranova3,4,5,6, Tatiana V Tatarinova6,7,8, Eleonora S Piruzian1 and Sergey Bruskin1,5 Psoriasis is a common inflammatory skin disease with complex etiology and chronic progression. To provide novel insights into the regulatory molecular mechanisms of the disease, we performed RNA sequencing analysis of 14 pairs of skin samples collected from patients with psoriasis. Subsequent pathway analysis and extraction of the transcriptional regulators governing psoriasis-associated pathways was executed using a combination of the MetaCore Interactome enrichment tool and the cisExpress algorithm, followed by comparison to a set of previously described psoriasis response elements. A comparative approach allowed us to identify 42 core transcriptional regulators of the disease associated with inflammation (NFκB, IRF9, JUN, FOS, SRF), the activity of T cells in psoriatic lesions (STAT6, FOXP3, NFATC2, GATA3, TCF7, RUNX1), the hyper- proliferation and migration of keratinocytes (JUN, FOS, NFIB, TFAP2A, TFAP2C) and lipid metabolism (TFAP2, RARA, VDR). In addition to the core regulators, we identified 38 transcription factors previously not associated with the disease that can clarify the pathogenesis of psoriasis. To illustrate these findings, we analyzed the regulatory role of one of the identified transcription factors (TFs), FOXA1. Using ChIP-seq and RNA-seq data, we concluded that the atypical expression of the FOXA1 TF is an important player in the disease as it inhibits the maturation of naive T cells into the (CD4+FOXA1+CD47+CD69+PD-L1(hi) FOXP3 − ) regulatory T cell subpopulation, therefore contributing to the development of psoriatic skin lesions.
    [Show full text]
  • (NGS) for Primary Endocrine Resistance in Breast Cancer Patients
    Int J Clin Exp Pathol 2018;11(11):5450-5458 www.ijcep.com /ISSN:1936-2625/IJCEP0084102 Original Article Impact of next-generation sequencing (NGS) for primary endocrine resistance in breast cancer patients Ruoyang Li1*, Tiantian Tang1*, Tianli Hui1, Zhenchuan Song1, Fugen Li2, Jingyu Li2, Jiajia Xu2 1Breast Center, Fourth Hospital of Hebei Medical University, Shijiazhuang, China; 2Institute of Precision Medicine, 3D Medicines Inc., Shanghai, China. *Equal contributors. Received August 15, 2018; Accepted September 22, 2018; Epub November 1, 2018; Published November 15, 2018 Abstract: Multiple mechanisms have been detected to account for the acquired resistance to endocrine therapies in breast cancer. In this study we retrospectively studied the mechanism of primary endocrine resistance in estrogen receptor positive (ER+) breast cancer patients by next-generation sequencing (NGS). Tumor specimens and matched blood samples were obtained from 24 ER+ breast cancer patients. Fifteen of them displayed endocrine resistance, including recurrence and/or metastases within 24 months from the beginning of endocrine therapy, and 9 pa- tients remained sensitive to endocrine therapy for more than 5 years. Genomic DNA of tumor tissue was extracted from formalin-fixed paraffin-embedded (FFPE) tumor tissue blocks. Genomic DNA of normal tissue was extracted from peripheral blood mononuclear cells (PBMC). Sequencing libraries for each sample were prepared, followed by target capturing for 372 genes that are frequently rearranged in cancers. Massive parallel
    [Show full text]