Arteriosclerosis, Thrombosis, and Vascular Biology

BRIEF REVIEW Proceedings of the Ninth HDL (High-Density ) Workshop Focus on

Annabelle Rodriguez, Bernardo L. Trigatti, Chieko Mineo, Darcy Knaack, John T. Wilkins, Daisy Sahoo, Bela F. Asztalos, Samia Mora, Marina Cuchel, Henry J. Pownall, Corina Rosales, Pascal Bernatchez, Amanda Ribeiro Martins da Silva, Godfrey S. Getz, Jacob L. Barber, Gregory C. Shearer, Angela M. Zivkovic, Uwe J.F. Tietge, Frank M. Sacks, Margery A. Connelly, Michael N. Oda, W. Sean Davidson, Mary G. Sorci-Thomas, Tomas Vaisar, Giacomo Ruotolo, Kasey C. Vickers, Catherine Martel

ABSTRACT: The HDL (high-density lipoprotein) Workshop was established in 2009 as a forum for candid discussions among academic basic scientists, clinical investigators, and industry researchers about the role of HDL in cardiovascular disease. This ninth HDL Workshop was held on May 16 to 17, 2019 in Boston, MA, and included outstanding oral presentations from established and emerging investigators. The Workshop featured 5 sessions with topics that tackled the role of HDL in the vasculature, its structural complexity, its role in health and disease states, and its interaction with the intestinal microbiome. The highlight of the program was awarding the Jack Oram Award to the distinguished professor emeritus G.S. Getz from the University of Chicago. The tenth HDL Workshop will be held on May 2020 in Chicago and will continue the focus on intellectually stimulating presentations by established and emerging investigators on novel roles of HDL in cardiovascular and noncardiovascular health and disease states. Downloaded from http://ahajournals.org by on October 22, 2019

Key Words: American Heart Association ◼ cardiovascular disease ◼ ◼ health ◼

he HDL (high-density lipoprotein) Workshop was HDL-C, statin treatment, systolic blood pressure, hyper- established in 2009 to advance our understanding tension treatment, history of diabetes mellitus, current Tof HDL biology.1 In clinical medicine, the measure- smoker, and aspirin therapy. Another well-known CVD ment of the cholesterol (C) component of HDL, HDL-C, risk calculator, the Framingham Risk Score, also incor- remains a standard-of-care requirement for cardiovas- porates HDL-C as a component in the 10-year risk cular disease (CVD) risk prediction. In 2019, the Ameri- assessment and assigns a score of −1 if patients have can Heart Association and other professional societies HDL-C≥60 mg/dL.4 issued an updated guideline on cholesterol manage- The latter would suggest that higher HDL-C levels ment for primary and secondary CVD prevention.2 This are cardioprotective, but this paradigm is now being chal- document targeted clinical management of LDL (low- lenged, and many argue for a reassessment of HDL-C in density lipoprotein)-C and non–HDL-C, while providing these risk calculators. Many clinical investigators agree no updates on HDL, HDL-C, or HDL functional metrics that the results of recent genetic association studies, as in CVD risk assessment. The American Heart Associa- well as large clinical trials targeting increases in HDL-C tion and the American College of Cardiology did provide to reduce CVD risk as a primary end point, have been an online risk calculator to help clinicians assess CVD disappointing.5–8 Such observations leave the field wide risk in their patients.3 The risk factors included in the open in better understanding the complexities of HDL calculator are sex, age, race, total cholesterol, LDL-C, biology and where this research might be of help to

Correspondence to: Annabelle Rodriguez, MD, Cell Biology, Linda and David Roth Chair of Cardiovascular Research, Center for Vascular Biology, Department of Cell Biology, University of Connecticut Health, 263 Farmington Ave, Farmington, CT 06030. Email [email protected] For Sources of Funding and Disclosures, see page xxx. © 2019 American Heart Association, Inc. Arterioscler Thromb Vasc Biol is available at www.ahajournals.org/journal/atvb

Arterioscler Thromb Vasc Biol. 2019;39:00–00. DOI: 10.1161/ATVBAHA.119.313340 December 2019 1 Rodriguez et al HDL Workshop

Nonstandard Abbreviations and Acronyms Highlights - VB - ApoA-I A-I • HDL (high-density lipoprotein) Workshop celebrates

eview CEC cholesterol efflux capacity its ninth year as a forum for academic basic scien- R CHD coronary heart disease tists, clinical investigators, and industry researchers to have candid discussions about HDL biology. rief CVD cardiovascular disease B • Presenters discussed novel aspects of scavenger EPA eicosapentaenoic acid receptors on cardiomyocyte function, LDL (low- EVs extracellular vesicles density lipoprotein) transendothelial cytosis, and FC free cholesterol raft distribution. HDL high-density lipoprotein • Presenters discussed many methodologies to evalu- ate the HDL proteome and association with cardio- HDL-p HDL particle vascular disease. JUPITER Justification for the Use of Statins in • Presenters discussed influences like serum opac- Prevention: an Intervention Trial Evaluat- ity factor, omega-3 fatty acids, egg consumption, ing Rosuvastatin exercise, and the intestinal microbiome on different LDL low-density lipoprotein aspects of HDL biology. LVs lymphatic vessels • G.S. Getz provided a comprehensive discussion PUFA polyunsaturated fatty acid about apoA-I and HDL proteome in oxLDL oxidized LDL as well as new in vivo approaches to assess HDL functionality. RCT reverse cholesterol transport SOF serum opacity factor SR-B1 scavenger receptor class B type I note that each of the following abstracts were written by WT wild-type the presenter with references included for the reader’s benefit. clinicians. A major objective of the HDL Workshop has been, and will continue to be, a forum for academic basic Session 1: HDL and the Vasculature Downloaded from http://ahajournals.org by on October 22, 2019 scientists, clinicians, and industry researchers to share 1. Bernardo L. Trigatti, PhD, Professor, Department of their research findings and perspectives. Over the past Biochemistry and Biomedical Sciences, McMaster Uni- 10 years attendance has steadily grown demonstrating versity and Thrombosis and Atherosclerosis Research that interest in HDL biology is alive and well. Therefore, Institute, McMaster University and Hamilton Health Sci- we now present the first of many future HDL Workshop ences, Hamilton, Ontario, Canada. HDL and SR-B1 in Proceedings with the hopes of engaging scientists and Cardiovascular Disease: Atherosclerosis and Beyond. clinicians to attend and present their research on this Global knockout of the HDL receptor, SR-B1 fascinating topic. (scavenger receptor class B type I (Scarb1 ), in ath- In this year’s program, the HDL Workshop took place erogenic mice increases their susceptibility to high-fat in Boston, MA, over a 2-day period. It included oral pre- diet induced aortic sinus atherosclerosis.9 In addition, sentations from invited established investigators as well when fed highly atherogenic, high fat and high choles- as short talks from trainees. A new feature was a poster terol diets, Scarb1/Ldlr double knockout mice develop presentation session. The highlight of the event was the occlusive coronary artery atherosclerosis, extensive annual Jack Oram Research Award, honoring the late Dr myocardial fibrosis, and early death.10 The extensive Jack Oram and generously funded by the Dyslipidemia myocardial fibrosis is undoubtedly triggered by the Foundation of Boston. This year’s award was presented occlusive coronary atherosclerosis in these mice, but to the distinguished professor emeritus Dr G.S. Getz from we wondered if it might also reflect a role for SR-B1 in the University of Chicago. cardiomyocytes themselves. SR-B1 is expressed in car- diomyocytes, and SR-B1’s ligand, HDL, is able to pro- tect cultured mouse and human cardiomyocytes from HDL WORKSHOP PROGRAM apoptosis induced by the anti-cancer drug doxorubicin.11 The program was chaired by Dr Catherine Martel and co- This protection is lost when SR-B1 is inactivated. HDL chaired by Dr Rodriguez. Over the course of the 2-day induces phosphorylation of Akt in mouse and human program, there were 5 sessions of oral presentations. cardiomyocytes in an SR-B1 dependent manner and The topics of each session were the following: HDL Akt1 or Akt2 knockdown impair HDL-mediated protec- and the Vasculature, HDL Structural Complexity, Clinical tion against doxorubicin mediated apoptosis.12 In vivo, Perspectives on HDL Measurement, HDL in Health and treatment of mice with doxorubicin over 5 weeks trig- Disease, and Microbiota and HDL Metabolism. Please gers greater cardiomyocyte apoptosis in Scarb1−/− than

2 December 2019 Arterioscler Thromb Vasc Biol. 2019;39:00–00. DOI: 10.1161/ATVBAHA.119.313340 Rodriguez et al HDL Workshop

wild-type (WT) mice. Transplantation studies implicate Lymphatic vessels (LVs) form a whole-body network B

SR-B1 in the heart itself. Finally, human apoA-I (apo- that maintains fluid balance, dietary lipid absorption, rief lipoprotein A-I) transgenic overexpression or injection and pathogen clearance from peripheral tissues. In the −/− R

protect WT but not Scarb1 mice against doxorubicin past years, the direct role of LVs in atherosclerosis has eview induced cardiomyocyte apoptosis and cardiac dysfunc- been more fully assessed. We reported that functional 11,12 tion. In conclusion, while global knockout studies adventitial LV are essential to first mobilize cholesterol - VB clearly demonstrate a protective role for SR-B1 against out of the vessel wall before reaching the circulation.16 atherosclerosis (likely by virtue of its function in A defect in the propelling capacity of the collecting LV, mediated reverse cholesterol transport [RCT]), SR-B1 rather than a defect in the absorptive capacity of the lym- and HDL appear to have cardioprotective roles beyond phatic capillaries would be the instigating element of this atherosclerosis. Notably, this includes protection of car- atherosclerosis-related lymphatic dysfunction.17 We are diomyocytes against cytotoxicity by the chemotherapeu- now pursuing the mechanisms that contribute to the loss tic agent doxorubicin. of collecting LV function during atherosclerosis, and how 2. Chieko Mineo, PhD, Associate Professor, Center for can they be modulated. Cell fragments called extracel- Pulmonary and Vascular Biology, Department of Pediat- lular vesicles (EVs) are important cell-cell messengers rics and Cell Biology, University of Texas Southwestern released on cell activation or death and are found in ath- Medical Center, Dallas, Texas. SR-B1 and Atherosclero- erosclerotic lesions. We reported for the first time that sis: A New Paradigm for an Old Story. EVs of heterogeneous origins are also present in lymph The biology of the high-affinity HDL receptor SR-B1 and that these small vesicles are even more abundant is best understood in the liver, where the receptor plays in atherosclerotic mice.18 With preliminary data suggest- a key role in RCT, facilitating cholesterol transfer from ing that specific subsets of EVs could be harmful to the HDL to hepatocytes for disposal in bile.13 The global lymphatic endothelium,19 we hypothesize that EVs accu- SR-B1 knockout mice show exaggerated atheroscle- mulation could be, at least in part, responsible for the rosis in the setting of hypercholesterolemia. In addition atherosclerosis-related lymphatic dysfunction. This mas- to the liver, SR-B1 is expressed in multiple cell types sive accumulation of EVs in the artery wall could, in turn, that are relevant to atherogenesis, including macro- be due to the underlying reduced LV function. Altogether, phages, platelets, and endothelium. In endothelial cells, these data suggest that EVs might be the connection SR-B1 has been shown to promote production of the between abnormal lymphatic function and atherosclero- Downloaded from http://ahajournals.org by on October 22, 2019 antiatherogenic molecule nitric oxide.14 To study how sis progression. endothelial SR-B1 impacts atherosclerosis, we gener- 4. Darcy Knaack, PhD, graduate student (Dr Daisy ated mice lacking SR-B1 selectively in the endothelium Sahoo, mentor), Department of Biochemistry, Division of by crossing floxed Scarb1 mice with VE-Cadherin Cre Endocrinology, Medical College of Wisconsin, Milwaukee, mice (Scarb1∆EC).15 Contrary to our prediction, Scarb1∆EC WI. The roles of SR-B1 and CD36 in Maintenance of Mac- mice were protected from hyperlipidemia-induced ath- rophage Cholesterol Homeostasis. erosclerosis without affecting the plasma lipid profiles. Atherosclerosis is a chronic inflammatory disease Recognizing that SR-B1 binds both HDL and LDL and characterized by buildup of cholesterol-rich macro- that LDL uptake into the vascular wall is the initial step phages within the endothelium. Two scavenger recep- of atherogenesis, we hypothesized that SR-B1 facili- tors that regulate macrophage cholesterol homeostasis tates LDL transport through the endothelial layer. Using and cholesterol transport are SR-B1 and CD36, which multiple loss-of-function approaches in vivo, we demon- bind HDL and oxLDL (oxidized LDL), respectively. Their strated that SR-B1 promotes LDL uptake into the aorta. individual roles in modulating atherosclerosis have been Studies in cultured endothelial cells further revealed that widely studied, but how they influence each other’s func- transendothelial transport of LDL requires SR-B1 C-ter- tions has yet to be investigated.20,21 To test our hypoth- minal cytoplasmic domain that recruits DOCK4. DOCK4 esis that SR-B1 and CD36 are functional partners that initiates LDL–SR-B1 internalization via activation of the mediate macrophage cholesterol homeostasis, we per- small GTPase Rac1. Our study revealed that expression formed 4 different assays in peritoneal macrophages of SR-B1 and DOCK4 is elevated in human atheroscle- from WT, Scarb1−/−, and Cd36−/− mice. First, coimmuno- rotic arteries compared with normal arteries. These find- precipitation assays demonstrated a potential interac- ings suggest that the mechanisms that influence LDL tion between SR-B1 and CD36 that was enhanced with transport across endothelium might be targeted to pro- oxLDL treatment in WT macrophages. Next, we exam- vide novel interventions to prevent atherosclerosis. ined how cholesterol transport functions of these recep- 3. Catherine Martel, PhD, Associate Professor, Can- tors were affected when one receptor was absent. We ada Research Chair in Lymphatics and Cardiovascular provide evidence that when CD36 is absent, the ability Medicine, Department of Medicine, Université de Mon- of HDL to bind surface receptors was impaired, as was tréal, Montreal Heart Institute, Montreal, Quebec, Canada. the ability of the cell ability to internalize HDL-cholesterol Extracellular Vesicles at the Heart of Lymphatic Function. as compared to WT or Scarb1−/− mice. We further show

Arterioscler Thromb Vasc Biol. 2019;39:00–00. DOI: 10.1161/ATVBAHA.119.313340 December 2019 3 Downloaded from http://ahajournals.org by on October 22, 2019 22, October on by http://ahajournals.org from Downloaded Brief Review - VB size subspecies. tors ofdifferent functionalcharacteristics across HDL apoA-I proteoformsmaybeimportant markersormedia- of apoA-IproteoformsinHDL-p issize-related andthat downstream proteomics.These datasuggest theprofile size separation technique and is compatible with HDL is anovel,high-resolution concluded thatCN-GELFrEE and 1.8×,respectively)inthemediumsizeranges.We significantly moreabundant(meanfolddifference: 1.7× respectively). Glycatedandoxidized proteoformswere types (7.5–9 nm; mean fold difference: 1.4× and 1.2×, nm) size ranges in comparison to medium size sub- 4 teome, lipidome, and functional characteristics. Particle SizeSubtypes. ferential ApoA-I Proteoform Quantification across HDL of Excellence,NorthwesternUniversity, Dif- Evanston,IL. of Life Processes Institute, and the Proteomics Center of Chemistry and Molecular Biosciences, the Chemistry versity, HenriqueSeckler, Departments Chicago,IL. MS, of Medicine,DivisionCardiology, NorthwesternUni- 1. JohnT. AssociateProfessor, Wilkins, MD, Department StructuralandtheVasculatureSession 2:HDL phage cholesterol homeostasis. suggesting theymayworktogethertopromotemacro- andCD36, be acooperativepartnershipbetweenSR-B1 domains. Basedontheseobservations,thereseemsto one receptorisabsent,theothermigratestononraft residewithinlipidraftmicrodomains,butwhen CD36 receptor. and Ourdatasuggestthat,inWTcells,SR-B1 impactsmembranelocalizationoftheother or CD36 fractionation toexamine whetherabsenceofSR-B1 either receptorisabsent.Last,weusedsucrose-gradient that membranedistributionofcholesterol isalteredwhen Rodriguez etal K88 apoA-Iinthepre-β K88 tion revealedasignificantlyhigherproportionofAcyl- (5–11nm).Proteoform quantifica- size rangeofHDL sizefractions covering the distinctHDL observed 36 mass spectrometryforproteoformquantification. We apoA-I-containing fractionstoliquid-chromatography size oftheelectrophoreticfractionsandthensubmitted ern blottoquantifyapoA-Icontentandaverageparticle technique, toseparateHDL-p bysize.We usedWest- anativeacrylamide-gelelectrophoretic CN-GELFrEE, Development inYoung Adultsparticipantsweadapted healthyCoronaryArteryRisk pooled samplesfrom30 types havedistinctapoA-Iproteoformprofiles.Using vary with HDL-p size. Wesub- hypothesized thatHDL efflux,butitisunknownifapoA-Iproteoforms HDL apoA-I)areassociatedwith forms ofApoA-I(Acyl-K88 (proteoforms) ofapoA-I. residues, yielding14distinctspecificmolecularforms lational modificationsthatoccuratpreciseaminoacid recently reportedthatapoA-Ihasspecificpost-trans- HDL particle (HDL-p) size subtypes differ in pro- HDL December 2019 23 -1 (5–7.5 nm) andα Fatty acid-modifiedproteo- -1 (9–11 Arterioscler ThrombVasc 10.1161/ATVBAHA.119.313340 DOI: Biol. 2019;39:00–00. 22 We We tect againstCVD. paralleling thefailureofHDL-raising therapeuticstopro- (SCARB1gene)mutationshavehigherCVD risk, SR-B1 commonandrare humansharboring levels ofHDL-C, Paradoxically, toSR-B1. of HDL despitehighcirculating tion viaRCT. The requiresthebinding laststepinRCT in deliveringperipheralcholesterol totheliverforexcre- Dr JekyllandMrHyde. kee, WI.The importance of HDL clearance: lessons from of Endocrinology, Medical College of Wisconsin, Milwau- (CE andTG).(CE andfree cholesterol/ The phospholipid/CE in thediscoid-shapedpre-β -1 particleswerecorelipids respectively. Interestingly, one-third ofthelipidmolecules inα-4,and257and 112inα-3,296 inpre-β-1, molecules were4and426 in α-1,3and227-2,2 functionality. The average numberofapoA-Iandlipid and controlsubjectshaving eitherloworhighpre-β-1 1-concentration-normalized cholesterol effluxcapacity) having low pre-β-1 functionality (as defined by pre-β- patients isolated fromcoronaryheartdisease(CHD) subpopulations 8 lipidclassesin5different-size HDL by theirlipidcomposition.We determinedapoA-Iand the functionalityofHDL-p issignificantlyinfluenced is poorly understood. We tested the hypothesis that on HDLcompositionandfunction. Research Center,Tufts University, Boston,MA.Analyses andreduceCVDenhance RCT risk. structurewillhelpusdevelopstrategiesto ing ofSR-B1 cholesterol transport. Together, an improved understand- mediates membrane/receptorinteractionstofacilitate testing whetherthisregionisajuxtamembranehelixthat electron paramagnetic resonance spectroscopy, we are intocells.Using bindinganddeliveryofHDL-C HDL exhibits impaired hydrophobic faceinfull-lengthSR-B1, a shortamphipathicα-helixthat,whenmutatedalongits peptide,wehaveidentified copy structureofanSR-B1 recently solvednuclearmagneticresonancespectros- tion canenhancecholesterol clearanceviaRCT. Inour interac- approach tounderstandhowtheSR-B1-HDL tookastructure-function these findings,ourlaboratory rophages togeneratereactiveoxygen species.Given increasedthe abilityofmac- malondialdehyde-HDL primary murine peritoneal macrophages. Furthermore, haveimpairedabilitiestopromotemigrationof HDL acrolein-andmalondialdehyde-modified to nativeHDL, inmacrophages.Compared protective functionsofHDL as acroleinandmalondialdehydeimpairedtheathero- modifiedbyreactivealdehydessuch observed thatHDL dysfunction throughincreasedoxidative modification. We clearancepromotesHDL hypothesis thatimpairedHDL We testedthe taining theprotectivefunctionsofHDL. that efficient cholesterol fluxRCT via is critical to main- HDL protectsagainstCVD, primarilyduetoitsrole HDL 2. Daisy Sahoo, PhD, Professor of Medicine, Division The composition andfunctionalrelationshipofHDL-p 3. BelaAsztalos,PhD,ScientistI,HumanNutrition 5–8,24,25 Thus, thefieldisappreciating HDL Workshop HDL Brief Review - VB 5 Excitingly, Excitingly, 33 HDL Workshop HDL-p might HDL-p 28 December 2019 26,27 Unfortunately, it is not available in Unfortunately, 31,32 Furthermore, HDL-C Furthermore, is not an adequate 29,30 The optimal range for a low risk of atherosclerotic The - MD, Pro Associate Cuchel, Marina 2. Research PhD, levels of HDL-CLow are associated with increased thanks to the greater awareness of its remarkable het- erogenicity in size and composition, our understanding of and diseases many of physiopathology the in role HDL’s beyond its role in the progres- conditions is expanding sion of atherosclerosis and RCT. also be of potential value in targeting residual CVD par- CVD residual targeting in value potential of be also prevention as indicated by data from ticularly for primary of JUPITER,the rosuvastatin arm but not where HDL-p, HDL-C,of CVD. was inversely predictive routine clinical care, and its results may be affected by be affected routine clinical care, and its results may and an as diabetes mellitus underlying conditions, such a method to assess mac- inflammatory status. Recently, rophage-specific developed that in vivo has been RCT could be of use during drug development. biomarker of HDL bio- function, and several additional RCTbeing investigated. markers are currently is one of functions of HDL.the most important atheroprotective Cholesterol efflux capacity (CEC), an in vitro assay HDLof ability the assessing from cholesterol efflux to macrophages (the first step of has been inversely RCT), associated with both prevalence and incidence of CVD in several studies. Session 4: HDL in Health and Disease of Bioenergetics, and PhD, Professor 1. Henry J. Pownall, - of Bioenerget Corina Rosales PhD, Assistant Professor ics, Institute for Academic Medicine, Houston Method- HDL and TX. Cornell Medical College, Houston, ist, Weill serum opacity factor. is HDL-Chumans among CVD 60between and 80 fessor, Department of Medicine, Division of Translational Department of Medicine, Division of Translational fessor, of School Medicine and Human Genetics, Perelman PA. Philadelphia, Pennsylvania, of University Medicine, Clinical perspectives on HDL measurement. the HDL-C asso- to LDL-C, contrary risk. However, CVD is not linear and, in recent years, risk ciation with CVD the absence of several data have emerged highlighting risk, or even an increase in CVD protection against CVD, at high HDL-C concentrations, suggesting the presence HDL-C between curve U-shaped a of all-cause and mortality. CVD prevention. At present, it can be measured clinically measured clinically present, it can be At prevention. CVD spectroscopy (LabCorp) magnetic resonance by nuclear - Most stud (Quest Diagnostics). analysis and ion mobility nuclear magnetic resonance based on the ies have been Study from the Multi-Ethnic method. Data spectroscopy and the placeboof Atherosclerosis of the JUPITER arm Prevention: an Use of Statins in (Justification for the Rosuvastatin) showed that Evaluating Intervention Trial HDL-C after adjusting of CVD is no longer predictive remained inversely associated HDL-p while for HDL-p, after adjusting for HDL-C.with CVD 24,25 -mobility particles. The pre-β-1 phospholipid/CEparticles. The α-mobility 2. Samia Mora, MD, Director, Center for Lipid MetaboLipid MD,Center for Samia Mora, 2. - Director, HDL-C risk, is a strong inverse indicator of CVD While A clinical vignette was presented to highlight the case A clinical vignette was presented to highlight lomics, Associate Professor of Medicine, Department of Medicine, Department lomics, Associate Professor and Cardiovascu- of Medicine, Divisions of Preventive Hospital, Harvard lar Medicine, Brigham and Women’s Boston, MA. Clinical perspectives on HDL Medical School, measurement. benefit. Given the into clinical yet translated this has not heterogeneity of HDLextreme structure and function, content of HDLmeasuring only the cholesterol will, at best, only partially reflect the potential role of HDL in risk assessment and therapeutic drug development. CVD has led to interest in developing HDLThis metrics that of functions atheroprotective indicate the better might HDL. average measurements include HDL-p, Proposed size, subclasses, and functional assays. Of the alternate HDLin utility potential has HDL-p of number the metrics, 1. Annabelle Rodriguez, MD, Professor, Cell Biology, Cell Biology, 1. Annabelle Rodriguez, MD, Professor, Research, Chair of Cardiovascular Linda and David Roth of Connecticut University Biology, Center for Vascular High HDL-C Paradox: Connecticut. Health, Farmington, A Clinical Vignette. HDL-Cof a 52-year-old woman with elevated (110 mg/ mg/dL) levels with a posi- (170 dL) and elevated LDL-C patient had no The tive family history of premature CVD. smoking, or history of known personal history of CVD, was found diabetes mellitus, but on physical examination patient was reluctant to have bilateral carotid bruits. The primary care physi- to start statin therapy as she and her cian believed that the HDL-C levels would provide car- demonstrated the high HDL-C case dioprotection. This of in this group for more research and the need paradox patients to uncover the underlying genetic and nonge- risk. netic etiologies of increased CVD Session 3: Clinical on HDL Perspectives Measurement - in low pre-β-1 functional significantly higher ratio was of or absence the presence of independent subjects ity CHD. between positive correlations were strong There abundance of major functionality and the pre-β-1 particle not high pre-β-1-functionality but in subjects with CHD.β-1-functionality and in subjects with low-pre- The was significantly the large-HDL-p lipid composition of between the CHDdifferent In and the control groups. CHDTG/CE patients, the high and phospholipid/CE with increased functionality of ratios were associated caus- clear support not do data These large-HDL-p. between the lipid composition and ative relationships Considering these results, HDL-p. the functionality of the lipid- other factors, for example, we hypothesize that functionality. binding capacity of apoA-I, regulate HDL-p CE pre-β-1 compared higher in ratios were significantly with Biol. 2019;39:00–00. DOI: 10.1161/ATVBAHA.119.313340 Arterioscler Thromb Vasc et al Rodriguez

Downloaded from http://ahajournals.org by on October 22, 2019 Downloaded from http://ahajournals.org by on October 22, 2019 22, October on by http://ahajournals.org from Downloaded Brief Review - VB nant HDL-FC influx is proportional to HDL-FCnant HDL-FC influxisproportionalto content. mol% FCdrivesinflux;themagnitudeofrecombi- containing≥15 HDL cellular FCefflux,recombinant 64% to 94% in double-mutant mice. 2bmicetoup to muscles ofDuchenne orLimb-Girdle muscle lesionareajumping from1%to5%inskeletal wasting comparedtoWT, mice,with apoEorsingleMD 2b/apoE micecanshowseveremuscle or Limb-Girdle Scarb1 associated virusdeliveryofserumopacityfactorto 6 levelsin mice. and lowersplasmatotalHDL structure Bacterial serumopacityfactordisruptsHDL transfers totheliverwitht in mice the majority of HDL- and nascent (n)HDL-FC isfast,t=<5minutesandreversible; transfer fromHDL Our hypothesisissupportedbyotherobservations.FC FC bioavailabilityprevents/reversestheseabnormalities. high HDL-FC bioavailabilityandthatreductionofHDL- levelsaredueto ties associatedwithhighplasmaHDL abnormali- support ourhypothesesthatthemetabolic mice isfreecholesterol (FC)-rich. These observations functionhavenotbeenidentified.Scarb1 HDL levelsisunknownandinterventionsthatrescue HDL-C atherosclerotic CVD amongpatientswithhighplasma syndrome,thecauseof are associatedwithmetabolic ated withhigherrisk.Whereas levels lowplasmaHDL-C mg/dL withhigherandlowerconcentrationsassoci- Rodriguez etal the females areinfertile. fromScarb1 Notably, HDL abnormalities;theyareatherosusceptible,and metabolic levelspresentwithmultiple which havehighplasmaHDL and skeletalmusclehomeostasis. cholesterol-loweringnew linksbetweenLDL approaches Columbia, Canada.HDL,LDL,andMuscularDystrophy. Innovation Centre,St.Paul’s Hospital,Vancouver, British peutics, UniversityofBritishColumbia,Heart&Lung Department ofAnesthesiology, Pharmacology&Thera- levels. plasma HDL problemsduetohigh ability andcouldreducemetabolic reduction ofHDL-p numberreducesHDL-FC bioavail- ≈24 hours.Inconclusion,serumopacityfactor-mediated rescues fertility amongthefemale Scarb1 tissues. tion; overlongertimes,nHDL-derived FCappearsinall in thesemice. Together, these dataclearlyshowedan complete lossofambulation, aphenotypeneverreported the factthatmuscledamage canreach levels thatcause inactivation andhigh-fatdiets. levelsareincreased via APOEgene when non–HDL-C skeletal muscle wasting and even loss of ambulation type 2bmice),wehaveobserveddrasticallyworsened MD muscle degeneration(Duchenne andLimb-Girdle thatexhibit weakbutchronic muscular dystrophy(MD) occurs is poorly understood. Using genetic models of can in rare casescause rhabdomyolysis, but how this Lipid managementpharmacotherapyhashighlighted 2. Pascal Bernatchez, PhD,AssociateProfessor, December 2019 −/ 34 Whereas FC-free recombinant HDL supports recombinantHDL Whereas FC-free micereducesHDL-FC tonilandconstitutively 1/ 2<5 minwithoutesterifica- 40 DoubleDuchenne/apoE 39 For instance,statins 41 More interesting is −/− 36–38 micewithin Arterioscler ThrombVasc 10.1161/ATVBAHA.119.313340 DOI: Biol. 2019;39:00–00. Adeno- −/− mice, −/− 35

dynamic range. subclasses duetoitshighsensitivity, specificity, andwide is apowerfulapproach toaccurately differentiate HDL spectrometry-based targetedquantitativeproteomics their abundancevariedconsiderably. DenseHDL subclasses,but majority ofproteinsweresharedbyboth subclasses.The with highprecisiontodifferentiate HDL Reaction Monitoring.Bothmethodsworkedequallywell odologies, DataIndependentAcquisitionandParallel their proteomewasquantifiedusing2targetedmeth- tions bydiscontinuousdensityultracentrifugation,and HDL-p wereseparatedintoHDL subclassesof19apparentlyhealthyvolunteers. of HDL with changesin theHDLproteome. of exercise-inducedchanges incholesterolefflux capacity Association Carolina,Columbia,SC. University ofSouth Sarzynski, mentor).Department ofExerciseScience, 1. JacobL.Barber, PhD,Graduatestudent(DrMarkA. Metabolism andHDL Session 5:Microbiota which makesaccuratequantificationa challenge. HDL-p, andmostofthemarepresentinlowabundance, the proteinsmaybedifferentially localizedtodistinct protein, make up to 90% oftheproteincontent. protein, makeupto90% APOA1 andAPOA2, the 2mostabundantproteinsinlipo- However,HDL. have beenidentifiedasassociatedwith functionality. proteomeaffects its contribute tounderstandhowHDL subclassesproteome and may gies todifferentiate HDL and Parallel ReactionMonitoringareattractivestrate- serum amyloid A-4. Thus, DataIndependent Acquisition apolipoproteinE,and andapoC-III), apoC-II, Cs (apoC-I, While HDL protein J, andphosphatidylcholine-sterol acyltransferase. protein D,apolipoproteinA-IV, apolipoproteinH,apolipo- activity, such paraoxonase andapolipoproteinL1,apolipo- significantly enriched withproteinsrelatedtoantioxidant ety ofcardioprotectivefunctions. quantitative proteomics. High-density lipoproteinssubclassesmappingbytargeted Chemistry, UniversityofSaoPaulo, SaoPaulo, Brazil. student (DrGraziellaE.Ronsein,mentor),Instituteof ther investigation. which warrantsfur- inMD, ciated cholesterol metabolism data suggestaprofounddefect inLDL- andHDL-asso- assessed bynormal creatine kinaselevels. Together, our also notedeveninsettingsoflowmuscledamageas lism wasanalyzed,andHDL-related abnormalitieswere - plasmalipoproteinmetabo settings ofDuchenne MD, possess intrinsic toxic properties in muscles. In various may ened tissuesandevensuggestedthatnon–HDL-C and musclehomeostasisinalreadygeneticallyweak- unexpected linkedbetweenplasmalipoproteinlevels HDL isaheterogeneousparticleandlinkedtovari- HDL 3. AmandaRibeiroMartinsdaSilva,PhD,Graduate 2 wasenriched withAPOA2, apolipoprotein 45 We, therefore,quantifiedtheproteome 42 Closeto100proteins 2 andHDL HDL Workshop HDL 3 subfrac- 43 44 Thus, Mass 3 was Brief Review - VB 7 HDL Workshop Within the bili- Within December 2019 53,54 Our findings indicate that diet 52 Impact of intestinal microbiota on reverse cho- 3. Angela M. Zivkovic, PhD, Assistant Professor, PhD, Assistant Professor, 3. Angela M. Zivkovic, and is growing evidence that the composition There 4. Uwe J.F. Tietge, MD, PhD, Professor, Division of of Division MD, Tietge, PhD, Professor, 4. Uwe J.F. Mediating RCT is envisioned as a major antiathero- ary pathway, cholesterol can be secreted either directly cholesterol ary pathway, or after metabolic into bile acids. RCT conversion studies conditions: ±apoA-I, and ±ABCA1 expression by siRNA ±apoA-I, and ±ABCA1conditions: expression - using compartmen efflux was analyzed Tracer silencing. phospholipids of label into media efflux The tal modeling. presence dependent on the esterified) was (as media of both ABCA1 apoA-I and in concurrently resulted and d8-12-HETE.unesterified intracellular lowest the Oppo- or ABCA1 the absence of apoA-I resulted in high sitely, and little to no appearance intracellular label appearance of d8-12-HETE pool. Overall, in the media phospholipid an important role of HDLour results suggest in mediat- efflux. ing oxylipin University California Davis, Department of Nutrition, in HDL lipidome, glyco- Diet-induced changes Davis, CA. capacity. proteome, and functional determinants are important functional capacity of HDL-p of HDLof the protective effects But we do not in CVD. and function of yet know how to improve the composition HDL.- feed cross-over In 2 controlled, randomized-order, of short-term diet ing studies, we determined the effects HDLon diet long-term and function. and composition days of fast food versus Mediterranean diet widely Four HDLthe remodeled com- lipid dietary reflect to lipidome A2HSG, but not A1AT, apoA-I content position, altered apoC-III, and did not alter protein glycopro- apoE, SAA, except apoC-IIIfiles, sialylation. Individual HDL lipid spe- with CEC;cies and glycopeptides were highly correlated were the diet-induced compositional changes however, in HDLnot accompanied by changes CEC possibly due total:HDL in baseline to differences ratios, cholesterol composition. Four study duration, or gut microbiome how- per day, weeks of 2 whole eggs versus egg whites HDLincreased ever, HDLto unrelated CEC,was which or lipidomic ApoA-I content, particle size or distribution, compositional changes. can alter HDL days. High composition in as little as 4 interindividual variability in response even in a tightly con- lipid by baseline explained partly be may study, trolled Our results also indicate profiles and gut microbiome. that diet can alter HDL CEC, that HDL and lipids and glycoproteins are highly correlated with HDL function, providing evidence that dietary strategies are a promising for improving the functional capacity of HDL.approach Institutet, Stockholm, Karolinska Clinical Chemistry, Sweden. lesterol transport. lend strong data Available function of HDL-p. sclerotic at least in rodents, the bili- support to a concept, in which, of RCT while ary secretion pathway accounts for ≈75% the remaining 25% occurs from the nonbiliary pathway excretion. of transintestinal cholesterol Increased 50,51 We examined examined We Further, subjects Further, 49 46–48 Exercise is known Exercise 32 - and other polyun Eicosapentaenoate =0.78) among nonresponders. This pilot pilot nonresponders. This among =0.78) Family Study. Measurement of the efflux of Study. Family HDLstrong a as identified been recently has function 2. Gregory C. Shearer, PhD, Associate Professor, PhD, Associate Professor, 2. Gregory C. Shearer, Recent clinical trials demonstrate 4 g/d of eicosa- Recent clinical trials demonstrate 4 =0.03), while there was no change in HDL=0.03), while there was no change APOE P predictor of cardiovascular risk. of cardiovascular predictor 3-PUFA intake is associated with large increases in ω3-PUFA action in PUFA seek to explain We HDL-EPA-oxylipins. by testing whether action specifically, general, and EPA HDLoxylipins. Oxylipins cellular efflux of mediates the and by acting as an oxylipin are intracellular oxylipins HDL acceptor, signaling. For could regulate intracellular detection purposes, we used 12-HETE as an oxylipin - from arachido It is produced by 12-lipoxygenase tracer. hypothesized efflux of 12-HETE from activated nate. We macrophages would be apoA-I and ABCA1 dependent. - macrophages were provided d8-arachi RAW264.7 under 4 donate and stimulated with 100 ng/mL LPS changes in CEC and concomitant changes in the HDLin the CECin changes concomitant and changes nonrelated individuals from the proteome in 19 white, HERITAGE radiolabeled and BODIPY-labeled from J774 cholesterol B depleted plasma, along macrophages to apolipoprotein HDL untargeted with performed was profiling, proteome - 20 weeks of endurance exer at baseline and following CEC; on of regular exercise was no effect cise. There in improvements in training resulted exercise however, CEC or subjects (responders) and either no changes in 9 (nonresponders). A total subjects 10 CEC in decreased HDLknown 33 of sub- 19 all in were identified in abundance of overall changes were no jects. There any HDL following training. Changes in abun- proteins dance of 6 HDL proteins (Albumin, IGHA1, IGHG2, C6, SAA4, and APOE) were nominally negatively correlated among responders, in CEC.with changes Additionally, HDL APOE abundance was significantly decreased ( abundance (P study provided limited evidence for a potential role of of role potential for a limited evidence provided study select HDL in CEC. proteins as mediators of changes to needed are sample sizes larger with studies Future further alterations in the HDL examine how proteome are related to HDL function. State University, Pennsylvania Nutritional Sciences, The PA. Park, University saturated fatty acids in HDL function. is associated with a 25% risk pentaenoic acid (EPA) reduction for myocardial infarction. to effect HDLto effect the metabolism risk, however, and CVD of HDL on measures of exercise effects function and known. are less well composition Biol. 2019;39:00–00. DOI: 10.1161/ATVBAHA.119.313340 Arterioscler Thromb Vasc with low HDLthose than had a greater benefit from EPA with high HDL. is an ω3-polyunsaturated fatty acid EPA are strong reasons to associate its effects There (PUFA). with HDL: HDL HDL transport PUFAs; responds most to treatment; and HDLω3-PUFA than host more oxylipins are signaling metaboany other plasma pool (oxylipins - including EPA). lites derived from PUFAs, et al Rodriguez

Downloaded from http://ahajournals.org by on October 22, 2019 Downloaded from http://ahajournals.org by on October 22, 2019 22, October on by http://ahajournals.org from Downloaded Brief Review - VB absorption. availability ofbileacidsisimportantalsoforcholesterol fecal neutralsterolfractioncouldincrease,sincethe be enhanced.Asasecondaryeffect withinthe alsoRCT withinthefecalsequestrants RCT bileacidfractioncould side effects. Conceivably, withtheapplicationofbileacid erosclerotic intervention strategy with relatively limited sequestrants represent a classical successful antiath- if such resultscanbetranslated.Nevertheless,bileacid research inthehumansystemisrequiredtoinvestigate, the rodent-specificmuricholic acidswereaffected, more lesterol withinthefecal bileacidfraction. is increasedduetomoresecretionofRCT-relevant cho- acids. Interestingly, intheabsenceofamicrobiotaRCT teria andthusthecapacityforproducingsecondarybile intestinal microbiota.Germ-freemicelack intestinalbac- liver, productsofthe secondarybileacidsaremetabolic dependent cholesterol efflux. ter isthefindingthatplasminogenpromotesABCA1 Anexample of the lat- positional heterogeneityofHDL. formationandthecom- as aplatformproteinforHDL may bereconciledbythepossibilitythatapoA-Iserves in humans and the Hybrid Diversity Mouse Panel. This as acausalgenefromgenome-wideassociationstudies animal experiments, theAPOA1genedoesnotappear evidence foratheroprotectionbyapoA-Iderivedfrom tribution. Second,itwasnotedthatdespitethestrong 8 HDL based on the role of apoA-I in distinguishing between andapoA-Iin humansandmice, ences betweenHDL humans andmice. ApolipoproteinA-IandHDLheterogeneityin Chicago, IL. Biochemistry, MolecularBiology, UniversityofChicago, Pritzker Distinguished Professor, Departments Pathology, PhD,ChairmanEmeritus,DonaldN. Godfrey S.Getz,MD, AWARD JACK ORAMRESEARCH bile acids. actuallyoccursvia strated thatalargeproportionofRCT neutral sterolversusthefecal bileacidfractiondemon- macrophage-derived cholesterol tracerwithinthefecal distinguishing betweentherespectiveexcretion ofthe Rodriguez etal of FVB mice is enriched in apoA-II. The latterproteinof of FVBmiceis enriched in apoA-II. (3aminoaciddifferences). HDL ferences) andapoA-II apoA-I (2aminoaciddif- shows apolymorphismofboth fromthe2strains erosclerosis. The comparisonofHDLs shows limited or no effect of apoA-I deficiency on ath- and atheroresistantstrains,respectively. The latterstrain andFVBmice,atherosensitive comparison ofC57BL6 in HybridDiversityMousePanel isexemplified bythe The factthat apoA-Idoesnotappearasamajorgene small cholesterol-free phospholipidcontainingparticles. and 8inahumanapoA-Ibackground favoredHDL plasma. ChimericapoA-Icontainingmousehelices7 Dr Getzfocusedon3topics.The firstbeingdiffer - December 2019 2 (mouseonly)andHDL 55 While primarybileacidsareformedinthe 57 Plasminogenisfoundina 3 andHDL 56 Sincemostly 2 inhuman Arterioscler ThrombVasc 10.1161/ATVBAHA.119.313340 DOI: Biol. 2019;39:00–00. 2 dis- treatment guidelines thatspecificallyaddress theroleof American HeartAssociation andothersshouldissue immunology. cargo, andHDL paradox, nontraditional HDL perspectives ofhighHDL-C asadrugplatform,clinical andreproduction,HDL HDL biology. TheHDL tentativeprogramwill includetopicson of what is known about that will push the boundaries established investigators and emerging ones, with topics ers. The roster ofspeakerswillcontinuetobeinnovative Rodriguez, andtheco-chair will beDrKasey C.Vick- place onMay2020inChicago.The chair willbeDr molecules. of heterogenousHDL guish the clinical utility of HDL-p versus the lipid content this speakstoanunderlyingfrictionintheneeddistin- might bemoreinformativeinCVD riskreduction.Allof gest thatmeasurementofHDL-p versuscholesterol trial sug- clinical perspective, results from the JUPITER results thatneedtestinginlargerpopulations.From a interventions with exercise and diet, with intriguing early proteomewereexamined by tal effects ontheHDL is stilldefinedby cholesterol effluxassays.Environmen- proteome, andassociationwithfunctionalitythatlargely different methodologiestoexamine theirnumber, size, continued focuswasonthecompositionofHDL-p, from ers oflipidsignalingmolecules(ie,oxylipins). Anareaof lymphatic transport,andtheirroleastransporters/efflux- theirroleinenhancing toxin-induced damageviaSR-B1, p, including,forexample, theirroleinprotectionfrom We arecontinuingtodiscovernovelfunctionsofHDL- biology.scientists exploring newparadigmsofHDL entific backgrounds, there is a vibrant community of investigators andemergingonesfromavarietyofsci- for whatbiologicalpurpose(s). scientists and clinicians to now convincingly determine exists. Asimplestatementandachallenge forbasic HDL PERSPECTIVES . ferent celltypesfortheirdirectinteractionatthelevelof could alsobeusedtoencapsulateothercellsor2dif- subject tovariousglobalenvironments.The method homeostasis andgeneexpression inthemacrophage recovery ofthemacrophages for assessmentofsterol of macrophagesinanalginatecapsule,allowingforthe lesterol effluxwasdescribed,involvingtheencapsulation along withapoA-I.Last,anovelmethodofinvivocho- proteins byexamining thereleaseofotherHDL HDL with proteomicscouldbeusedtoprobethestructureof Advantage could be taken of this remodeling, combined ismuch morestabletothisremodeling. tides; FVBHDL can beremodeledbyexposure totandemmimeticpep- the 2strainsshowdifferences invivodynamics.HDL We thinkthathealthcareorganizationssuch asthe Workshop willtake Looking forward, thetenthHDL As evidencedbythetalentedrosterofestablished HDL Workshop HDL Brief Review - VB

- 9 HDL Workshop December 2019 mice. J.T. Wilkins declares having served declares having Wilkins mice. J.T. −/− https://hdlworkshop.org/. Accessed August 8, 2019. 2018 AHA/ACC/AACBPR/AAPA/ABC/ACPM/ADA/AGS/APhA/ASPC/ NLA/PCNA Circulation. Guideline on the management of blood cholesterol. 2019;139:e1082–e1143. Risk CV Guidelines Tool American Heart Association. 2018 Prevention http://static.heart.org/riskcalc/app/index.html#!/baseline-risk. Calculator. Accessed July 8, 2019. Hard Coronary Heart Disease (10-year risk). Heart Study. Framingham https://www.framinghamheartstudy.org/fhs-risk-functions/hard-coronary- heart-disease-10-year-risk/. Accessed July 8, 2019. GM, Peloso Orho-Melander M, Frikke-Schmidt R, Barbalic M, BF, Voight Jensen MK, et al. Plasma HDL Hindy G, Hólm H, Ding EL,- cho Johnson T, study. randomisation a mendelian myocardial infarction: and risk of lesterol doi: 10.1016/S0140-6736(12)60312-2 Lancet. 2012;380:572–580. GG, Abt M, Ballantyne CM,Schwartz Olsson AG, Barter PJ, Brumm J, Chaitman BR, Holme IM, LA, et al; dal-OUT D, Leiter Kallend COMES of dalcetrapib in patients with a recent Investigators. Effects acute coronary syndrome. N Engl J Med. 2012;367:2089–2099. doi: 10.1056/NEJMoa1206797 T, Chaitman Desvignes- BR, Boden WE, Probstfield JL, Anderson W; AIM-HIGH K, Weintraub K, McBride R, Teo Koprowicz P, Nickens with low HDLNiacin in patients Investigators. levels receiv- cholesterol doi: N Engl J Med. 2011;365:2255–2267. ing intensive statin therapy. 10.1056/NEJMoa1107579 Komajda M, Caulfield M, Eriksson M, Grundy Kastelein JJ, SM, Barter PJ, DD, et al; ILLUMINATE JC, Waters Lopez-Sendon J, Mosca L, Tardif 7. 2. 1. 3. 4. 5. 6. 8. Disclosures rights inventorship declares and Genomics Lipid of the founder is Rodriguez Dr declares inventorship rights to issued patents related to HDL Trigatti B.L. biology. to issued patents related to Scarb1 REFERENCES nadian Institutes of Health Research (MOP74753, PJT-156437, and PJT-162272) and PJT-162272) PJT-156437, (MOP74753, of Health Research nadian Institutes (G-16-00014064Canada of G-19- and Foundation and Heart the and supported by NIH C. Mineo was was supported 0026275). D. Knaack HL126795. Endowment. Healthier Wisconsin Center and the Advancing by the Cardiovascular grant support from the NIH/NHLBI received Wilkins J.T. K23HL133601-04. supported by NIHD. Sahoo was HL58012. supported by B. Asztalos was NIHof the National Institutes has received support from S. Mora HL117933. Health (HL134811, HL117861, HL136852, and DK112940). has M. Cuchel was supported by NIH H.J. Pownall NIHreceived support from the HL059407. sup- was Bernatchez P. NIH supported by was C. Rosales HL129767. HL129767. and the Jain Foundation. of Health Research ported by the Canadian Institutes HL131028.G.S. Getz was supported by by multiple J.L. Barber was supported grants from the NIHNIH/NIGMS to Dr Sarzynski: P20 GM103499 and NIH/ NHLBI R01HL146462. Rohatgi: NIH/NHLBI NIH/NHLBI R01HL136724, K08HL118131, The American Heart Association 15CVGPSD27030013. and HERITAGE Study: NIH Family Dr Gregory C. Shearer was supported HL45670. by HL130099 supported by USDA and institutional funds. A.M. Zivkovic was Na- project CA-D-NUT-2242-H and Hatch and Agriculture, tional Institute of Food NIH Davidson was supported by NIH grant R01 AG062240. W.S. P01HL128203 was supported by NIHgrant. M.G. Sorci-Thomas HL127649 HL38907 and P01HL092969, supported by NIH was Vaisar grants P30DK017047, grants. T. P01HL128203, was supported by NIH and R01HL144558. K.C. Vickers Awards HL128996, and HL116263. C. Martel as supported by grants from HL127173, (363262),the Canadian Institutes of Health Research the Natural Sciences and of Canada (RGPIN-2016-05331), Council Engineering Research the Canadian du Quebec - Santé - New de recherche for Innovation (35289, a Fond Foundation Chair 2 - Junior 1) and a Canada Research Scholars investigator award (Research and Cardiovascular Medicine. in Lymphatics as a consultant for NGM support Bio. S. Mora declares having received research Diagnostics and serving as a consultant to Quest Diagnostics from Atherotech declares receiving support for clinical trials unrelated to M. Cuchel and Pfizer. HDL Regeneron Pharmaceuticals, and Regenxbio. from Akcea Therapeutics, G.C. Shearer declares receiving C.Rosales was supported by NIH HL129767. Sacks Pharmaceuticals. F. speakership and advisory panel honoraria from Amarin M.A. Connelly is an em- declares being a consultant for Pfizer and AstraZeneca. lipoprotein profiling services via nuclear magnetic offers ployee of LabCorp, which resonance spectroscopy (NMR). M.N. Oda is the founder of DRx Inc. BioLogics, is a consultant for MedImmune/AstraZeneca. Dr Giacomo Ruotolo is an Vaisar T. no conflicts. other authors report The employee of Eli Lilly and Company. Department of Biochemistry and Bio- and Biochemistry of Department We think it is important to expand into other funda- into other to expand think it is important We urgent and this collaborative clinical need is The Sources of Funding of Sources Dr Rodriguez has been supported by a National Institutes of Health (NIH) HL131862Linda and David Roth Chair of grant and an endowment from the has received funding support from the Ca- B.L. Trigatti Cardiovascular Research. From the Center for Vascular Biology, Department of Cell Biology, University of Department of Cell Biology, Biology, the Center for Vascular From (A.R.); Farmington Health, Connecticut Re- and Thrombosis and Atherosclerosis medical Sciences, McMaster University, University and Hamilton Health Sciences, Hamilton, Institute, McMaster search Department of Biology, and Vascular Center for Pulmonary ON, Canada (B.L.T.); Southwestern Medical Center, University of Texas Cell Biology, and Pediatrics of Endocri- D.S.) and Division (D.K., of Biochemistry Dallas (C.M.); Department Milwaukee; Division of Medical College of Wisconsin, nology (D.S., M.G.S.-T.), Northwestern Medicine, Preventive of and Medicine of Departments Cardiology, University, Tufts Center, Research Nutrition Human IL Chicago, (J.T.W.); University, for Lipid Metabolomics, and Center of Preventive Divisions (B.F.A.); Boston, MA Hospi- Women’s Brigham and Medicine, Department of Medicine, Cardiovascular Medicine Boston, MA (S.M.); Division of Translational tal, Harvard Medical School, of Medicine, University of Pennsylvania, School and Human Genetics, Perelman Houston Methodist, Weill Philadelphia (M.C.); Institute for Academic Medicine, C.R.); Department of Anesthesiol- (H.J.P., Cornell Medical College, Houston, TX University of British Columbia, Heart and Pharmacology and Therapeutics, ogy, Institute BC, Canada (P.B.); Hospital, Vancouver, Innovation Centre, St Paul’s Lung - Brazil (A.R.M.d.S.); Department of Pathol University of Sao Paulo, of Chemistry, ILof Chicago, University Science, Univer- of Exercise Department (G.S.G.); ogy, of Southsity The Sciences, Nutritional Department (J.L.B.); Columbia Carolina, Nutrition, of Department (G.C.S.); Park University University, State Pennsylvania of Department Chemistry, of Clinical Division (A.M.Z.); Davis California University Clini- Sweden (U.J.F.T.); Laboratory Institutet, Stockholm, Karolinska Medicine, University Hospital, Karolinska University Laboratory, Karolinska cal Chemistry, Health, Boston, of Public Chan School Harvard T.H. Sweden (U.J.F.T.); Stockholm, Laboratory Corporation of America Holdings (LabCorp), Morrisville, MA (F.M.S.); Pathol- CA (M.N.O.); Department of NC (M.A.C.); DRx Inc, Fairfield, BioLogics, UW Medicine ogy and Laboratory Medicine, University of Cincinnati, OH (W.S.D.); Eli Lilly and Company, (T.V.); Seattle Diabetes Institute, University of Washington, Indianapolis, IN of Cardiovascular Medicine, Department of Medi- (G.R.); Division and Montreal of Medicine, Nashville, TN (K.C.V.); School University cine, Vanderbilt of Medicine, Uni- Heart Institute, Montreal and Department of Medicine, Faculty versité de Montréal, Montreal, QC, Canada (C.M.). ARTICLE INFORMATIONARTICLE 2019. Received August 14, 2019; accepted September 20, Affiliations HDL-Cand risk at patients of management clinical the in evidence shows that overwhelming The CVD. those with low HDL-C risk CVD value for increased adds predictive HDL-Cwhile high for cardio- not show evidence does about- and func of HDL-p the question What protection. to the clinical trials do point Results from large tionality? assays as pro- efflux and cholesterol possibility of HDL-p more much However, reduction. risk CVD in utility viding to determine if these assays can work needs to be done to relevant manner throughput high a in performed be insurance reimbursement. clinical medicine and our program of HDLaspects as shown by mental biology, - tenth HDLdevelopment for the explo These Workshop. us into causal pathways influ- rations will likely guide by HDLenced directly or secondarily that will be targets applications. for future clinical diagnostic and therapeutic and indus- effort of academic basic scientists, clinicians, find the answers to why will certainly try researches HDL exists. Biol. 2019;39:00–00. DOI: 10.1161/ATVBAHA.119.313340 Arterioscler Thromb Vasc et al Rodriguez

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