Abbreviations:ANGPTL (angiopoietin-like protein), ApoA5 (apolipoprotein A-V), ASCVD (atherosclerotic cardiovascular disease), CAD (coronary artery disease), CI (confidence interval), EL (endothelial lipase), GWAS (genome-wide association study), HL (hepatic lipase), HMGCR (HMG-CoA reductase), IV (instrumental variable), LD (linkage disequilibrium), LDLR (LDL receptor), LIPC (lipase C hepatic type), LIPG (lipase G endothelial type), LPL (lipoprotein lipase), NES (normalized effect size), NMR (nuclear magnetic resonance spectroscopy), PCSK9 (proprotein convertase subtilisin/kexin type 9), R2 (coefficient of determination)
- Ginsberg H.N.
- Packard C.J.
- Chapman M.J.
- Boren J.
- Aguilar-Salinas C.A.
- Averna M.
- et al.
- Ginsberg H.N.
- Packard C.J.
- Chapman M.J.
- Boren J.
- Aguilar-Salinas C.A.
- Averna M.
- et al.
- Georgiadi A.
- Lichtenstein L.
- Degenhardt T.
- Boekschoten M.V.
- van Bilsen M.
- Desvergne B.
- et al.
MATERIALS AND METHODS
Study design and data sources
|Derivation cohort||Validation set|
|Data source||UK Biobank (UKBB)||Meta-analysis of 11 European studies|
|Study design||Population-based cohort||Population-based cohorts, case-control studies (only controls used), birth cohort, family-based studies (adjusted for family structure), twin studies (adjusted for relatedness)|
|Population||British||Dutch, Estonian, Finnish, German|
|Sample size per metabolic parameter||110,058 – 115,078||13,171 – 24,925|
|Genotyping platform(s)||UK Biobank axiom array / IDT xGen Exome Research Panel v1.0 w. Illumina NovaSeq 6000 platform||Affymetrix 6.0, Affymetrix 250K, Affymetrix 6.0 907K, Illumina 318K, Illumina 370K, Illumina 610K, Illumina 660K, Illumina 670K, Illumina CoreExome, Illumina Human660W, Illumina HumanCNV370, Illumina HumanOmniExpress, Illumina Omni 1M, Illumina OmniExpress, Perlegen-Affymetrix 500K|
|Number of SNPs||12,321,875 / 2,043,019||11,274,684 – 12,092,490|
|Metabolic parameter analysis platform(s)||500 MHz Bruker AVANCE III HD||500 MHz Bruker AVANCE III HD, 600 MHz Bruker AVANCE II|
|Number of metabolic parameters||248||122|
Genetic instrument justification
Validation of genetic instruments
|Protein target||Genetic variant used for instrumentation (chr:pos)||Allele frequency (derivation/validation set)||Mutation consequence||Effect on clinical phenotypes|
|LPL||rs115849089-A (8:19,912,370)||11.4 – 11.5 % /10.0 – 11.2 %||Increases the expression of LPL: Located ∼153,000 base pairs downstream of the LPL gene. Expression quantitative trait loci (eQTL) for LPL RNA expression in whole blood (Figure S1).||TG: -0.18; ≤ 2×10-308|
HDL: 0.16; ≤ 2×10-308
LDL: -0.01; 6×10-5
CAD: 0.95; 4×10-9
|ANGPTL3||rs11207977-T (1:62,977,307)||35.0 – 35.1 % /26.9 – 29.0 %||Decreases the expression of ANGPTL3: DOCK7 intronic variant located ∼86,000 base pairs upstream of ANGPTL3. eQTL for ANGPTL3 RNA expression in hepatic tissue (Figure S4).||TG: -0.08; 2×10-304|
HDL: -0.02; 2×10-13
LDL: -0.04; 7×10-69
CAD: 0.99; 0.69
|ANGPTL4||rs116843064-A (19:8,429,323)||2.0 % /2.9 – 3.1 %||Alters the function of ANGPTL4: Missense variant that causes the E40K substitution. This destabilizes the protein after secretion and prevents ANGPTL4 inhibition of LPL (|
|TG: -0.22; 1×10-16|
HDL: 0.21; 3×10-195
LDL: 0; 0.9
CAD: 0.87; 4×10-10
|ANGPTL8||rs2278426-T (19:11,350,488)||3.5 % /5.7 – 6.5 %||Alters the function of ANGPTL3/ANGPTL8 complex: Missense variant that causes a R59W replacement. In vitro studies suggest ANGPTL3-ANGPTL8 R59W have a decreased ability to bind and inhibit LPL, when compared to wild-type ANGPTL3-ANGPTL8 (||TG: -0.04; 1×10-14|
HDL: -0.09; 3×10-69
LDL: -0.04; 4×10-15
CAD: 1.01; 0.25
|ANGPTL8||rs145464906-T (19:11,350,874)||0.04 %||Protein-truncating variant: Nonsense Q121X mutation that results in a premature stop codon. Truncated ANGPTL8 does not form complex with ANGPTL3 (|
|TG: -0.54; 2×10-33|
HDL: 0.44; 3×10-22
LDL: -0.15; 2×10-3
CAD: 0.73; 0.21
|EL (LIPG)||rs77960347-G (18:47,109,955)||1.3 % /0.5 – 0.7 %||Alters the function of EL: Missense mutation causing the N396S substitution. Leads to a 40 % decrease of EL activity in vitro (|
|TG: 0.03; 5×10-4|
HDL: 0.29; 4×10-269
LDL: 0.07; 8×10-15
CAD: 0.90; 9×10-5
|HL (LIPC)||rs1800588-T (15:58,723,675)||21.4 – 21.6 % /23.5 – 25.0 %||Decreases the expression of HL: Promoter variant associated with ∼ 30% lower LIPC promoter activity in cell lines and ∼ 15% lower HL activity in human post-heparin plasma (|
|TG: 0.05; 2×10-84|
HDL: 0.12; ≤ 2×10-308
LDL: 0.02; 5×10-17
CAD: 1.03; 2×10-5
LPL mimicry patterns of ANGPTL3, ANGPTL4, and ANGPTL3/ANGPTL8 complexes
Multivariable LPL mimicry analysis using ANGTPL3- and ANGPTL8 variants
Genetic evidence for ANGPTL3/ANGPTL8 complex inhibition of EL
ANGPTL8 protein-truncating variant shows very high concordance with LPL enhancement
The LPL-ANGPTL3 residual variance is explained by ANGPTL3 action through EL
No evidence for ANGPTL4 influence on plasma lipids through HL or EL
DATA AVAILABILITY STATEMENT
- Triglyceride-rich lipoproteins and their remnants: metabolic insights, role in atherosclerotic cardiovascular disease, and emerging therapeutic strategies-a consensus statement from the European Atherosclerosis Society.European heart journal. 2021; 42: 4791-4806
- Physiological regulation of lipoprotein lipase.Biochim Biophys Acta. 2014; 1841: 919-933
- New insights into angiopoietin-like proteins in lipid metabolism and cardiovascular disease risk.Curr Opin Lipidol. 2019; 30: 205-211
- An updated ANGPTL3-4-8 model as a mechanism of triglyceride partitioning between fat and oxidative tissues.Prog Lipid Res. 2022; 85: 101140
- Characterization of the fasting-induced adipose factor FIAF, a novel peroxisome proliferator-activated receptor target gene.J Biol Chem. 2000; 275: 28488-28493
- Peroxisome proliferator-activated receptor gamma target gene encoding a novel angiopoietin-related protein associated with adipose differentiation.Mol Cell Biol. 2000; 20: 5343-5349
- Absence of ANGPTL4 in adipose tissue improves glucose tolerance and attenuates atherogenesis.JCI insight. 2018; : 3
- Angiopoietin-like 4 directs uptake of dietary fat away from adipose during fasting.Molecular metabolism. 2017; 6: 809-818
- Role of Kupffer cells in the pathogenesis of liver disease.World J Gastroenterol. 2006; 12: 7413-7420
- Transgenic angiopoietin-like (angptl)4 overexpression and targeted disruption of angptl4 and angptl3: regulation of triglyceride metabolism.Endocrinology. 2005; 146: 4943-4950
- Linking nutritional regulation of Angptl4, Gpihbp1, and Lmf1 to lipoprotein lipase activity in rodent adipose tissue.BMC physiology. 2012; 12: 13
- Angptl4 upregulates cholesterol synthesis in liver via inhibition of LPL- and HL-dependent hepatic cholesterol uptake.Arterioscler Thromb Vasc Biol. 2007; 27: 2420-2427
- The Fasting-induced Adipose Factor/Angiopoietin-like Protein 4 Is Physically Associated with Lipoproteins and Governs Plasma Lipid Levels and Adiposity.J Biol Chem. 2006; 281: 934-944
- Regulation of plasma triglyceride partitioning by adipose-derived ANGPTL4 in mice.Scientific reports. 2021; 11: 7873
- ANGPTL4 mediates shuttling of lipid fuel to brown adipose tissue during sustained cold exposure.eLife. 2015; : 4
- Fatty acid-inducible ANGPTL4 governs lipid metabolic response to exercise.Proc Natl Acad Sci U S A. 2014; 111: E1043-E1052
- ANGPTL4 deficiency in haematopoietic cells promotes monocyte expansion and atherosclerosis progression.Nature communications. 2016; 7: 12313
- Induction of cardiac Angptl4 by dietary fatty acids is mediated by peroxisome proliferator-activated receptor beta/delta and protects against fatty acid-induced oxidative stress.Circ Res. 2010; 106: 1712-1721
- Angptl4 protects against severe proinflammatory effects of saturated fat by inhibiting fatty acid uptake into mesenteric lymph node macrophages.Cell Metab. 2010; 12: 580-592
- Characterization of ANGPTL4 function in macrophages and adipocytes using Angptl4-knockout and Angptl4-hypomorphic mice.J Lipid Res. 2019; 60: 1741-1754
- Angiopoietin-like 4 promotes intracellular degradation of lipoprotein lipase in adipocytes.J Lipid Res. 2016; 57: 1670-1683
- Angiopoietin-like 4 promotes the intracellular cleavage of lipoprotein lipase by PCSK3/furin in adipocytes.J Biol Chem. 2018; 293: 14134-14145
- The angiopoietin-like protein ANGPTL4 catalyzes unfolding of the hydrolase domain in lipoprotein lipase and the endothelial membrane protein GPIHBP1 counteracts this unfolding.eLife. 2016; : 5
- Hepatocyte-specific suppression of ANGPTL4 improves obesity-associated diabetes and mitigates atherosclerosis in mice.J Clin Invest. 2021;
- Inactivating Variants in ANGPTL4 and Risk of Coronary Artery Disease.N Engl J Med. 2016; 374: 1123-1133
- ANGPTL8 protein-truncating variant associated with lower serum triglycerides and risk of coronary disease.PLoS Genet. 2021; 17: e1009501
- Coding Variation in ANGPTL4, LPL, and SVEP1 and the Risk of Coronary Disease.N Engl J Med. 2016; 374: 1134-1144
- Population-based resequencing of ANGPTL4 uncovers variations that reduce triglycerides and increase HDL..Nat Genet. 2007; 39: 513-516
- Genetics of blood lipids among ∼300,000 multi-ethnic participants of the Million Veteran Program.Nat Genet. 2018; 50: 1514-1523
- Association of Genetically Enhanced Lipoprotein Lipase-Mediated Lipolysis and Low-Density Lipoprotein Cholesterol-Lowering Alleles With Risk of Coronary Disease and Type 2 Diabetes.JAMA cardiology. 2018; 3: 957-966
- Angptl3 regulates lipid metabolism in mice.NatGenet. 2002; 30: 151-157
- Fasting-induced adipose factor/Angiopoietin-like protein 4: a potential target for dyslipidemia.Future Lipidology. 2006; 1: 227-236
- ANGPTL3 decreases very low density lipoprotein triglyceride clearance by inhibition of lipoprotein lipase.J Biol Chem JID - 2985121R. 2002; 277: 33742-33748
- Genetic and Pharmacologic Inactivation of ANGPTL3 and Cardiovascular Disease.N Engl J Med. 2017; 377: 211-221
- Variants with large effects on blood lipids and the role of cholesterol and triglycerides in coronary disease.Nat Genet. 2016; 48: 634-639
- A highly conserved motif within the NH2-terminal coiled-coil domain of angiopoietin-like protein 4 confers its inhibitory effects on lipoprotein lipase by disrupting the enzyme dimerization.J Biol Chem. 2009; 284: 11942-11952
- Atypical angiopoietin-like protein that regulates ANGPTL3.Proc Natl Acad Sci U S A. 2012; 109: 19751-19756
- Identification of RIFL, a novel adipocyte-enriched insulin target gene with a role in lipid metabolism.Am J Physiol Endocrinol Metab. 2012; 303: E334-E351
- Lipasin, a novel nutritionally-regulated liver-enriched factor that regulates serum triglyceride levels.Biochem Biophys Res Commun. 2012; 424: 786-792
- ANGPTL8 promotes the ability of ANGPTL3 to bind and inhibit lipoprotein lipase.Molecular metabolism. 2017; 6: 1137-1149
- ANGPTL8 requires ANGPTL3 to inhibit lipoprotein lipase and plasma triglyceride clearance.J Lipid Res. 2017; 58: 1166-1173
- ANGPTL8 has both endocrine and autocrine effects on substrate utilization.JCI insight. 2020; : 5
- Angiopoietin-like protein 8 differentially regulates ANGPTL3 and ANGPTL4 during postprandial partitioning of fatty acids.J Lipid Res. 2020; 61: 1203-1220
- Angiopoietin-like protein 3 governs LDL-cholesterol levels through endothelial lipase-dependent VLDL clearance.J Lipid Res. 2020; 61: 1271-1286
- LDL-Cholesterol Reduction by ANGPTL3 Inhibition in Mice Is Dependent on Endothelial Lipase.Circ Res. 2020; 127: 1112-1114
- Angiopoietin-like protein 4 (ANGPTL4) is an inhibitor of endothelial lipase (EL) while the ANGPTL4/8 complex has reduced EL-inhibitory activity.Heliyon. 2021; (e07898): 7
- Angiopoietin-like 3 inhibition of endothelial lipase is not modulated by angiopoietin-like 8.J Lipid Res. 2021; 62: 100112
- Metabolomic Profiling of Statin Use and Genetic Inhibition of HMG-CoA Reductase.J Am Coll Cardiol. 2016; 67: 1200-1210
- Metabolomic consequences of genetic inhibition of PCSK9 compared with statin treatment.Circulation. 2018; 138: 2499-2512
- Metabolic profiling of angiopoietin-like protein 3 and 4 inhibition: a drug-target Mendelian randomization analysis.Eur Heart J. 2021; 42: 1160-1169
- Characterising metabolomic signatures of lipid-modifying therapies through drug target mendelian randomisation.PLoS Biol. 2022; 20: e3001547
- Genome-wide study for circulating metabolites identifies 62 loci and reveals novel systemic effects of LPA.Nat Commun. 2016; 7: 11122
- Genetic effects on gene expression across human tissues.Nature. 2017; 550: 204-213
- Discovery and refinement of loci associated with lipid levels.Nat Genet. 2013; 45: 1274-1283
- Assessing the contribution of rare-to-common protein-coding variants to circulating metabolic biomarker levels via 412,394 UK Biobank exome sequences.medRxiv. 2021; 2021 (12.24.21268381)
- Evaluating the relationship between circulating lipoprotein lipids and apolipoproteins with risk of coronary heart disease: A multivariable Mendelian randomisation analysis.PLoS Med. 2020; 17: e1003062
- Identification of 64 Novel Genetic Loci Provides an Expanded View on the Genetic Architecture of Coronary Artery Disease.Circ Res. 2018; 122: 433-443
- Whole-exome imputation within UK Biobank powers rare coding variant association and fine-mapping analyses.Nat Genet. 2021; 53: 1260-1269
- UK biobank: an open access resource for identifying the causes of a wide range of complex diseases of middle and old age.PLoS Med. 2015; 12: e1001779
- Quantitative serum nuclear magnetic resonance metabolomics in cardiovascular epidemiology and genetics.Circ Cardiovasc Genet. 2015; 8: 192-206
- Quality control and removal of technical variation of NMR metabolic biomarker data in ∼120,000 UK Biobank participants.medRxiv. 2021; 2021 (09.24.21264079)
- Davey Smith G, Ala-Korpela M. Quantitative Serum Nuclear Magnetic Resonance Metabolomics in Large-Scale Epidemiology: A Primer on -Omic Technologies.Am J Epidemiol. 2017; 186: 1084-1096
- Bayesian test for colocalisation between pairs of genetic association studies using summary statistics.PLoS Genet. 2014; 10: e1004383
- LDlink: a web-based application for exploring population-specific haplotype structure and linking correlated alleles of possible functional variants.Bioinformatics. 2015; 31: 3555-3557
- Homozygosity for two point mutations in the lipoprotein lipase (LPL) gene in a patient with familial LPL deficiency: LPL(Asp9-->Asn, Tyr262-->His).J Lipid Res. 1996; 37: 651-661
- Genetic variation in ANGPTL4 provides insights into protein processing and function.J Biol Chem. 2009; 284: 13213-13222
- Human genomics. Effect of predicted protein-truncating genetic variants on the human transcriptome.Science. 2015; 348: 666-669
- On the mechanism of angiopoietin-like protein 8 for control of lipoprotein lipase activity.J Lipid Res. 2019; 60: 783-793
- Loss-of-function variants in endothelial lipase are a cause of elevated HDL cholesterol in humans.J Clin Invest. 2009; 119: 1042-1050
- The C-514T polymorphism in the human hepatic lipase gene promoter diminishes its activity.J Lipid Res. 2000; 41: 155-158
- Common C-to-T substitution at position -480 of the hepatic lipase promoter associated with a lowered lipase activity in coronary artery disease patients.Arterioscler Thromb Vasc Biol. 1997; 17: 2837-2842
- Association of variation in hepatic lipase activity with promoter variation in the hepatic lipase gene. The LOCAT Study Invsestigators.J Clin Invest. 1998; 101: 956-960
- Equivalence of the mediation, confounding and suppression effect.Prev Sci. 2000; 1: 173-181
- Angiopoietin-like protein3 regulates plasma HDL cholesterol through suppression of endothelial lipase.Arterioscler Thromb Vasc Biol. 2007; 27: 366-372
- Hepatic proprotein convertases modulate HDL metabolism.Cell Metab. 2007; 6: 129-136
- An anti-ANGPTL3/8 antibody decreases circulating triglycerides by binding to a LPL-inhibitory leucine zipper-like motif.J Lipid Res. 2022; : 100198
- Effect of Vupanorsen on Non-High-Density Lipoprotein Cholesterol Levels in Statin-Treated Patients With Elevated Cholesterol: TRANSLATE-TIMI 70. Circulation. 2022;
- Evinacumab for Homozygous Familial Hypercholesterolemia.N Engl J Med. 2020; 383: 711-720
- Highly accurate protein structure prediction with AlphaFold.Nature. 2021; 596: 583-589
- The Arg59Trp variant in ANGPTL8 (betatrophin) is associated with total and HDL-cholesterol in American Indians and Mexican Americans and differentially affects cleavage of ANGPTL3.Mol Genet Metab. 2016; 118: 128-137
- Genetic and Metabolic Determinants of Plasma Levels of ANGPTL8.J Clin Endocrinol Metab. 2021; 106: 1649-1667
- Association of low-frequency and rare coding-sequence variants with blood lipids and coronary heart disease in 56,000 whites and blacks.Am J Hum Genet. 2014; 94: 223-232
- Within-sibship GWAS improve estimates of direct genetic effects.bioRxiv. 2021; 2021 (03.05.433935)
- The MRC IEU OpenGWAS data infrastructure.bioRxiv. 2020; 2020 (08.10.244293)
Publication stageIn Press Accepted Manuscript
Funding sources: This research was funded by grants to FL from the Västerbotten Regional Council (RV-970117).
Fredrik Landfors: Conceptualization, Methodology, Software, Validation, Formal analysis, Investigation, Resources, Data curation, Writing – Original Draft, Writing – Review & Editing, Visualization. Elin Chorell: Resources, Writing – Review & Editing, Supervision, Project administration, Funding acquisition. Sander Kersten: Conceptualization, Writing – Original Draft, Writing – Review & Editing, Supervision.
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