Rare variant in scavenger receptor BI raises HDL cholesterol and increases risk of coronary heart disease

TLDR: In this paper, the authors identified a homozygote for a loss-of-function variant, in which leucine replaces proline 376 (P376L), in SCARB1, the gene encoding SR-BI.

Abstract: Scavenger receptor BI (SR-BI) is the major receptor for high-density lipoprotein (HDL) cholesterol (HDL-C). In humans, high amounts of HDL-C in plasma are associated with a lower risk of coronary heart disease (CHD). Mice that have depleted Scarb1 (SR-BI knockout mice) have markedly elevated HDL-C levels but, paradoxically, increased atherosclerosis. The impact of SR-BI on HDL metabolism and CHD risk in humans remains unclear. Through targeted sequencing of coding regions of lipid-modifying genes in 328 individuals with extremely high plasma HDL-C levels, we identified a homozygote for a loss-of-function variant, in which leucine replaces proline 376 (P376L), in SCARB1, the gene encoding SR-BI. The P376L variant impairs posttranslational processing of SR-BI and abrogates selective HDL cholesterol uptake in transfected cells, in hepatocyte-like cells derived from induced pluripotent stem cells from the homozygous subject, and in mice. Large population-based studies revealed that subjects who are heterozygous carriers of the P376L variant have significantly increased levels of plasma HDL-C. P376L carriers have a profound HDL-related phenotype and an increased risk of CHD (odds ratio = 1.79, which is statistically significant).

Figures

Fig. 2. SCARB1 P376L is a null variant in vitro and in vivo (A) [3H] Cholesterol ether (CEt) uptake (left) and selective cholesterol uptake from HDL (right) in iPSC-derived HLCs from the P376L homozygote versus a noncarrier control. Cells were incubated with [3H]CEt and 125I-labeled tyramine cellobiose (TC) dual-labeled human

Fig. 1. HDL composition and functionality in a SCARB1 P376L homozygote, heterozygous carriers, and controls (A) FPLC fractionation of plasma lipoproteins from the P376L homozygote subject (red) and from a control with normal HDL-C. (B) Cholesterol, apoA-I, and apoA-II content in total HDL. (C) Free cholesterol (FC) and esterified cholesterol (CE) in total HDL (left) and the FC/CE ratio in total HDL (right). (D) HDL subclass concentrations after separation by

Full text

Rare variant in scavenger receptor BI raises HDL
cholesterol and increases risk of coronary heart disease
Item Type Article
Authors Zanoni, P.; Khetarpal, S. A.; Larach, D. B.; Hancock-Cerutti,
W. F.; Millar, J. S.; Cuchel, M.; DerOhannessian, S.; Kontush,
A.; Surendran, P.; Saleheen, D.; Trompet, S.; Cooper, R. S.;
Chowdhury, R.; Dedoussis, G.; de Faire, U.; Feranil, A. B.; Sheu,
W. H.- H.; Ferrucci, L.; Freimer, N. B.; Gieger, C.; Sengupta, S.;
Grallert, H.; Groop, L. C.; Gudnason, V.; Tanaka, T.; Gyllensten,
U.; Hamsten, A.; Erdmann, J.; Harris, T. B.; Shuldiner, A. R.;
Hingorani, A.; Hirschhorn, J. N.; Gravito, M. L.; Wilson, J. F.;
Hofman, A.; Hovingh, G. K.; Hsiung, C. A.; Humphries, S. E.;
Teslovich, T. M.; Hunt, S. C.; Hveem, K.; Iribarren, C.; Siegbahn,
A.; Groves, C. J.; Nordestgaard, B. G.; Njolstad, I.; Jarvelin, M.-
R.; Jula, A.; Kahonen, M.; Kaprio, J.; Kesaniemi, A.; Kivimaki,
M.; Thorleifsson, G.; Kooner, J. S.; Hallmans, G.; Koudstaal,
P. J.; Kuulasmaa, K.; Gustafsson, S.; Krauss, R. M.; Kuh, D.;
Nielsen, S. F.; Kuusisto, J.; Kyvik, K. O.; Laakso, M.; Lakka, T. A.;
Hartikainen, A.-L.; Lind, L.; Van den Herik, E. G.; Lindgren, C. M.;
Kanoni, S.; Spector, T. D.; Martin, N. G.; Marz, W.; McCarthy, M.
I.; McKenzie, C. A.; Tybjaerg-Hansen, A.; Assimes, T. L.; Meneton,
P.; Metspalu, A.; Moilanen, L.; Morris, A. D.; Ganna, A.; Voight,
B. F.; Stefansson, K.; Munroe, P. B.; van Pelt, L. J.; Waite, L. L.;
Hayward, C.; Strachan, D. P.; Tayo, B. O.; Tremoli, E.; Tuomilehto,
J.; Uusitupa, M.; Chen, J.; van Duijn, C. M.; Vollenweider, P.;
Wallentin, L.; Liu, D. J.; Hernandez, D.; Vedantam, S.; Wareham,
N. J.; Jukema, J. W.; Whitfield, J. B.; Wolffenbuttel, B. H. R.;
Ordovas, J. M.; Buchkovich, M. L.; Boerwinkle, E.; Palmer, C.
N. A.; Thorsteinsdottir, U.; Hicks, A. A.; Chasman, D. I.; Rotter,
J. I.; Wainwright, N.; Franks, P. W.; Perola, M.; Wong, A.; Riatti,
S.; Mora, S.; Cupples, L. A.; Sandhu, M. S.; Rauramaa, R.; Rich,
S. S.; Boehnke, M.; Deloukas, P.; Mohlke, K. L.; Wijmenga, C.;
Ingelsson, E.; Gu, D.; Roberts, R.; Beckmann, J. S.; Wu, Y.; Peloso,
G. M.; Blankenberg, S.; Watkins, H.; Blankenberg, S.; Clarke, R.;
Collins, R.; Kim, B.-J.; Wild, S. H.; McPherson, R.; Nieminen, M.
S.; Barroso, I.; Holm, H.; O'Donnell, C.; Schreiber, S.; Zhang, W.;
Siegbahn, A.; Salomaa, V.; Zalloua, P. A.; Mannisto, S.; Amouyel,

P.; Willemsen, G.; Arveiler, D.; Hung, Y.-J.; Bragg-Gresham,
J. L.; Ferrieres, J.; Muller-Nurasyid, M.; Ferrario, M.; Kee, F.;
Absher, D.; Willer, C. J.; Samani, N.; Schunkert, H.; Butterworth,
A. S.; Sattar, N.; Wilsgaard, T.; Chang, H.-Y.; Howson, J. M. M.;
Pedersen, N. L.; Di Angelantonio, E.; Demirkan, A.; Den Hertog, H.
M.; Do, R.; Donnelly, L. A.; Ehret, G. B.; Illig, T.; Esko, T.; Feitosa,
M. F.; Ferreira, T.; Been, L. F.; Power, C.; Fischer, K.; Song,
C.; Fontanillas, P.; Fraser, R. M.; Freitag, D. F.; Jones, M. R.;
Gurdasani, D.; Heikkila, K.; Hypponen, E.; Isaacs, A.; Jackson,
A. U.; Pramstaller, P. P.; Johansson, A.; Bolton, J. L.; Young, E.
H.; Johnson, T.; Kaleebu, P.; Kaakinen, M.; Kettunen, J.; Kleber,
M. E.; Li, X.; Luan, J.; Lyytikainen, L.-P.; Price, J. F.; Magnusson,
P. K. E.; Mangino, M.; Mihailov, E.; Kastelein, J. J. P.; Zhao, J.
H.; Bonnycastle, L. L.; Montasser, M. E.; Nolte, I. M.; OConnell,
J. R.; Palmer, C. D.; Petersen, A.-K.; Schmidt, R. F.; Sanna, S.;
Saxena, R.; Bandinelli, S.; Service, S. K.; Shah, S.; Adair, L. S.;
Shungin, D.; Brambilla, P.; Sidore, C.; De Craen, A.; Burnett, M.
S.; Psaty, B. M.; Cesana, G.; Khaw, K.-T.; Dimitriou, M.; Doney, A.
S. F.; Doring, A.; Elliott, P.; Arveiler, D.; Epstein, S. E.; Eyjolfsson,
G. I.; Gigante, B.; Goodarzi, M. O.; Quertermous, T.; Kim, E.;
Deloukas, P.; Stitziel, N. O.; Volcik, K. A.; Klopp, N.; Komulainen,
P.; Kumari, M.; Langenberg, C.; Ford, I.; Lehtimaki, T.; Lin, S.-Y.;
Lindstrom, J.; Bennett, F.; Danesh, J.; Loos, R. J. F.; Uitterlinden,
A. G.; Mach, F.; McArdle, W. L.; Meisinger, C.; Mitchell, B. D.;
Muller, G.; Nagaraja, R.; Packard, C.; Narisu, N.; Kathiresan, S.;
Nieminen, T. V. M.; Bochud, M.; Salomaa, V.; Nsubuga, R. N.;
Olafsson, I.; Ong, K. K.; Palotie, A.; Papamarkou, T.; Pomilla, C.;
Pouta, A.; Rader, D. J.; Reilly, M. P.; Majumder, A. a. S.; Ridker,
P. M.; Sanghera, D. K.; Boehm, B. O.; Rivadeneira, F.; Rudan, I.;
Ruokonen, A.; Scharnagl, H.; Seeley, J.; Asiki, G.; Silander, K.;
Stancakova, A.; Stirrups, K.; Swift, A. J.; Saramies, J.; Alam, D.
S.; Boomsma, D. I.; Tiret, L.; Strawbridge, R. J.; Abecasis, G.;
Watson, S.; Borecki, I. B.; Bornstein, S. R.; Bovet, P.; Burnier, M.;
Campbell, H.; Schwarz, P. E. H.; Chakravarti, A.; Chambers, J. C.;
Chen, Y.-D. I.; Collins, F. S.; Schmidt, E. M.; Surakka, I.
Citation Rare variant in scavenger receptor BI raises HDL cholesterol and
increases risk of coronary heart disease 2016, 351 (6278):1166
Science
DOI 10.1126/science.aad3517
Publisher American Association for the Advancement of Science
Journal Science

Rights Copyright © 2016, American Association for the Advancement of
Science.
Download date 10/08/2022 01:36:17
Item License http://rightsstatements.org/vocab/InC/1.0/
Version Final accepted manuscript
Link to Item http://hdl.handle.net/10150/623258

Rare variant in scavenger receptor BI raises HDL cholesterol and
increases risk of coronary heart disease
Paolo Zanoni
1,*
, Sumeet A. Khetarpal
1,*
, Daniel B. Larach
1,*
, William F. Hancock-Cerutti
1,2
,
John S. Millar
1
, Marina Cuchel
1
, Stephanie DerOhannessian
1
, Anatol Kontush
2
, Praveen
Surendran
3
, Danish Saleheen
3,4,5
, Stella Trompet
6,7
, J. Wouter Jukema
7,8
, Anton De
Craen
6
, Panos Deloukas
9
, Naveed Sattar
10
, Ian Ford
11
, Chris Packard
12
, Abdullah al Shafi
Majumder
13
, Dewan S. Alam
14
, Emanuele Di Angelantonio
3
, Goncalo Abecasis
15
, Rajiv
Chowdhury
3
, Jeanette Erdmann
16
, Børge G. Nordestgaard
17
, Sune F. Nielsen
17
, Anne
Tybjærg-Hansen
18
, Ruth Frikke Schmidt
19
, Kari Kuulasmaa
20
, Dajiang J. Liu
21
, Markus
Perola
20,22
, Stefan Blankenberg
23,24
, Veikko Salomaa
20
, Satu Männistö
20
, Philippe
Amouyel
25
, Dominique Arveiler
26
, Jean Ferrieres
27
, Martina Müller-Nurasyid
28,29
, Marco
Ferrario
30
, Frank Kee
31
, Cristen J. Willer
32
, Nilesh Samani
33,34
, Heribert Schunkert
35
, Adam
S. Butterworth
3
, Joanna M. M. Howson
3
, Gina M. Peloso
36
, Nathan O. Stitziel
37
, John
Danesh
3,9
, Sekar Kathiresan
36
, and Daniel J. Rader
1,†
CHD Exome+ Consortium
‡
Consortium CARDIoGRAM Exome, Consortium Global Lipids Genetics
1
Departments of Genetics and Medicine, Division of Translational Medicine and Human Genetics,
Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
2
INSERM
UMR 1166 ICAN, Université Pierre et Marie Curie Paris 6, Hôpital de la Pitié, Paris, France
3
Cardiovascular Epidemiology Unit, Department of Public Health and Primary Care, University of
Cambridge, Cambridge, UK
4
Department of Biostatistics and Epidemiology, Perelman School of
Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
5
Centre for Non-
Communicable Diseases, Karachi, Pakistan
6
Department of Gerontology and Geriatrics, Leiden
University Medical Center, Leiden, Netherlands
7
Department of Cardiology, Leiden University
Medical Center, Leiden, Netherlands
8
The Interuniversity Cardiology Institute of the Netherlands,
Utrecht, Netherlands
9
Wellcome Trust Sanger Institute, Genome Campus, Hinxton, UK
10
Institute
of Cardiovascular and Medical Sciences, British Heart Foundation, Glasgow Cardiovascular
Research Centre, University of Glasgow, Glasgow, UK
11
Robertson Center for Biostatistics,
University of Glasgow, Glasgow, UK
12
Glasgow Clinical Research Facility, Western Infirmary,
Glasgow, UK
13
National Institute of Cardiovascular Diseases, Sher-e-Bangla Nagar, Dhaka,
Bangladesh
14
International Centre for Diarrhoeal Disease Research, Mohakhali, Dhaka,
†
Corresponding author. ; Email: rader@mail.med.upenn.edu
*
These authors contributed equally to this work.
‡
For each consortium and study, authors and affiliations are listed in the supplementary materials.
SUPPLEMENTARY MATERIALS
www.sciencemag.org/content/351/6278/1166/suppl/DC1
Materials and Methods
Supplementary Text
Figs. S1 to S7
Table S1
References (40–76)
Consortia and Study Author Lists
Published in final edited form as:
Science
. 2016 March 11; 351(6278): 1166–1171. doi:10.1126/science.aad3517.

Bangladesh
15
Center for Statistical Genetics, Department of Biostatistics, University of Michigan
School of Public Health, Ann Arbor, MI 48109, USA
16
Institute for Integrative and Experimental
Genomics, University of Lübeck, Lübeck 23562, Germany
17
Department of Clinical Biochemistry,
Herlev Hospital, Copenhagen University Hospital, Herlev, Denmark
18
Copenhagen University
Hospital, University of Copenhagen, Copenhagen, Denmark
19
Department of Clinical
Biochemistry, Rigshospitalet, Copenhagen University Hospitals, Copenhagen, Denmark
20
Department of Health, National Institute for Health and Welfare, Helsinki, Finland
21
Department
of Public Health Sciences, College of Medicine, Pennsylvania State University, Hershey, PA
17033, USA
22
Institute of Molecular Medicine FIMM, University of Helsinki, Helsinki, Finland
23
Department of General and Interventional Cardiology, University Heart Center Hamburg,
Hamburg, Germany
24
University Medical Center Hamburg-Eppendorf, Hamburg, Germany
25
Department of Epidemiology and Public Health, Institut Pasteur de Lille, Lille, France
26
Department of Epidemiology and Public Health, University of Strasbourg, Strasbourg, France
27
Department of Epidemiology, Toulouse University-CHU Toulouse, Toulouse, France
28
Institute of
Genetic Epidemiology, Helmholtz Zentrum München–German Research Center for Environmental
Health, Neuherberg, Germany
29
Department of Medicine I, Ludwig-Maximilians-University
Munich, Munich, Germany
30
Research Centre in Epidemiology and Preventive Medicine,
Department of Clinical and Experimental Medicine, University of Insubria, Varese, Italy
31
UKCRC
Centre of Excellence for Public Health, Queens University, Belfast, Northern Ireland
32
Department
of Computational Medicine and Bioinformatics, Department of Human Genetics, and Department
of Internal Medicine, University of Michigan, Ann Arbor, MI 48109, USA
33
Department of
Cardiovascular Sciences, University of Leicester, Leicester, UK
34
National Institute for Health
Research (NIHR) Leicester Cardiovascular Biomedical Research Unit, Glenfield Hotel, Leicester,
UK
35
Deutsches Herzzentrum München, Technische Universität München, Munich, Germany
36
Broad Institute and Center for Human Genetic Research, Massachusetts General Hospital,
Boston, MA 02114, USA
37
Department of Medicine, Division of Cardiology, Department of
Genetics, and the McDonnell Genome Institute, Washington University School of Medicine, St.
Louis, MO 63110, USA
Abstract
Scavenger receptor BI (SR-BI) is the major receptor for high-density lipoprotein (HDL)
cholesterol (HDL-C). In humans, high amounts of HDL-C in plasma are associated with a lower
risk of coronary heart disease (CHD). Mice that have depleted Scarb1 (SR-BI knockout mice)
have markedly elevated HDL-C levels but, paradoxically, increased atherosclerosis. The impact of
SR-BI on HDL metabolism and CHD risk in humans remains unclear. Through targeted
sequencing of coding regions of lipid-modifying genes in 328 individuals with extremely high
plasma HDL-C levels, we identified a homozygote for a loss-of-function variant, in which leucine
replaces proline 376 (P376L), in
SCARB1
, the gene encoding SR-BI. The P376L variant impairs
posttranslational processing of SR-BI and abrogates selective HDL cholesterol uptake in
transfected cells, in hepatocyte-like cells derived from induced pluripotent stem cells from the
homozygous subject, and in mice. Large population-based studies revealed that subjects who are
heterozygous carriers of the P376L variant have significantly increased levels of plasma HDL-C.
Zanoni et al. Page 2

Frequently asked questions

Q1. What are the contributions in "Rare variant in scavenger receptor bi raises hdl cholesterol and increases risk of coronary heart disease" ?

Zanoni et al. this paper identified a homozygote for a loss-of-function variant, in which leucine replaces proline 376 ( P376L ), in SCARB1, encoding SR-BI. 

Q2. What is the role of SR-BI in the cell surface?

Given that SR-BI undergoes N-glycosylation in the endoplasmic reticulum concomitant with proper folding, the authors hypothesized that altered posttranslational modification may underlie its reduced cell surface localization (27–29). 

Q3. What is the significance of the lower-molecular weight forms?

Higher-molecular-weight forms represent N-glycosylation modified EndoH–resistant and partially sensitive forms at the cell surface after modification by alphamannosidase II in the Golgi apparatus (28). 

Q4. What is the effect of SR-BI in the liver?

In mice, overexpression of SR-BI in the liver reduces levels of HDL-C (7–10), and genetic deletion of SR-BI results in higher HDL-C levels (11–13). 

Q5. What is the role of SR-BI in HDL cholesterol metabolism?

SR-BI promotes the selective uptake of HDL cholesteryl esters (HDL-CEs) into cells, particularly hepatocytes and steroidogenic cells (5, 6). 

Q6. Why did the authors find the markedly reduced HDL-CE uptake in the iPS?

The authors hypothesized that the markedly reduced HDL-CE uptake could be because of aberrant processing of the P376L SR-BI protein, which leads to impaired cell surface localization. 

Q7. How did the authors sequence the exons of SR-BI in mice?

In this cohort, the authors sequenced the exons of ~990 genes located within 300 kb of each of the 95 loci with significant associations (P < 5 × 10−8) with plasma lipid levels identified by the Global Lipids Genetics Consortium as of 2010 (22). 

Q8. What is the significance of SR-BI in human physiology?

Studies of injected labeled HDL-CE in humans suggested that the majority of the HDL-CE was transported to the liver via CETPmediated exchange to apoB-containing lipoproteins rather than by direct uptake from HDL by the liver (30), which brings into question the importance of hepatic SR-BI in human physiology. 

Q9. What is the effect of the P376L variant on HDL-C levels?

To evaluate the physiological impact of the P376L variant on HDL-C levels and catabolism in vivo, the authors used adeno-associated virus (AAV) vectors to direct hepatic overexpression of WT SR-BI or the P376L variant in mice with depleted Scarb1 [Scarb1 knockout (KO) mice]. 

Q10. What is the important finding of this study?

Perhaps the most important finding of their study is that, despite the elevation in HDL-C, P376L carriers exhibit increased risk of CHD, as do Scarb1 KO mice. 

Q11. What is the sensitivity of the lysates of different genotypes of SR?

Cell lysates of each genotype were treated with Endo-H to remove complex N-linked glycans from mature forms of proteins and then immunoblotted for SR-BI. 

Q12. What is the role of SR-BI in the development of atherosclerosis?

SCARB1 P376L is associated with increased risk of CHD in humansDespite a profound increase in HDL-C, SR-BI deficiency in mice causes accelerated atherosclerosis (17–20). 

Q13. What is the effect of genetic manipulations on atherosclerosis in mice?

these genetic manipulations in mice have effects on atherosclerosis opposite to those predicted by human epidemiological data: Overexpression reduces atherosclerosis despite the lower HDL-C levels (14–16), and gene deletion increases atherosclerosis despite the higher HDLC levels (17–20). 

Q14. What is the effect of the P376L variant on SR-BI?

This indicates that the P376L sequence variant results in complete loss of the canonical function of SR-BI—namely, selective uptake of HDL-CE. 

Q15. What is the role of SR-BI in the Golgi?

Page 5the endogenous posttranslational N-glycosylation of SR-BI to prevent either transit from the ER to the Golgi or further posttranslational modifications in the Golgi, which ultimately result in reduced cell surface expression. 

Q16. What is the effect of SR-BI on HDL-C?

the variant retains substantial SR-BI activity, no homozygotes were identified, the apparent effect on HDL-C was modest, and there was insufficient power to address its effects on atherosclerosis. 

Q17. What grants were awarded to support the recruitment of patients?

This work was supported in part by an award from the National Center for Research Resources (grant TL1RR024133) and National Center for Advancing Translational Sciences of the NIH (grant TL1R000138) to support patient recruitment.