. 2018 Jun 13;9(1):2252.

doi: 10.1038/s41467-018-04611-z.

Genetic inactivation of ANGPTL4 improves glucose homeostasis and is associated with reduced risk of diabetes

Viktoria Gusarova¹, Colm O'Dushlaine², Tanya M Teslovich², Peter N Benotti³, Tooraj Mirshahi³, Omri Gottesman², Cristopher V Van Hout², Michael F Murray³, Anubha Mahajan⁴, Jonas B Nielsen^{5

6}, Lars Fritsche⁷, Anders Berg Wulff⁸, Daniel F Gudbjartsson⁹, Marketa Sjögren¹⁰, Connor A Emdin¹¹, Robert A Scott¹², Wen-Jane Lee^{13

14}, Aeron Small^{15

16}, Lydia C Kwee¹⁷, Om Prakash Dwivedi¹⁸, Rashmi B Prasad¹⁹, Shannon Bruse², Alexander E Lopez², John Penn², Anthony Marcketta², Joseph B Leader³, Christopher D Still³, H Lester Kirchner³, Uyenlinh L Mirshahi³, Amr H Wardeh³, Cassandra M Hartle³, Lukas Habegger², Samantha N Fetterolf³, Teresa Tusie-Luna^{20

21}, Andrew P Morris^{4

22

23}, Hilma Holm⁹, Valgerdur Steinthorsdottir⁹, Patrick Sulem⁹, Unnur Thorsteinsdottir⁹, Jerome I Rotter²⁴, Lee-Ming Chuang^{25

26}, Scott Damrauer^{27

28}, David Birtwell^{15

16}, Chad M Brummett²⁹, Amit V Khera^{11

30}, Pradeep Natarajan^{11

30}, Marju Orho-Melander¹⁰, Jason Flannick^{11

31}, Luca A Lotta¹², Cristen J Willer^{5

6

32}, Oddgeir L Holmen³³, Marylyn D Ritchie³, David H Ledbetter³, Andrew J Murphy¹, Ingrid B Borecki², Jeffrey G Reid², John D Overton², Ola Hansson^{18

19}, Leif Groop^{18

19}, Svati H Shah¹⁷, William E Kraus¹⁷, Daniel J Rader^{15

16}, Yii-Der I Chen²⁴, Kristian Hveem^{33

34

35}, Nicholas J Wareham¹², Sekar Kathiresan³⁰, Olle Melander¹⁰, Kari Stefansson⁹, Børge G Nordestgaard^{36

37

38

39}, Anne Tybjærg-Hansen^{8

36

38

39}, Goncalo R Abecasis⁷, David Altshuler^{11

40

41

42}, Jose C Florez^{43

44

45}, Michael Boehnke⁷, Mark I McCarthy^{4

46

47}, George D Yancopoulos¹, David J Carey³, Alan R Shuldiner², Aris Baras⁴⁸, Frederick E Dewey⁴⁹, Jesper Gromada⁵⁰

Affiliations

¹ Regeneron Pharmaceuticals, Tarrytown, 10591, NY, USA.
² Regeneron Genetics Center, Tarrytown, 10591, NY, USA.
³ Geisinger, Danville, 17822, PA, USA.
⁴ Wellcome Centre for Human Genetics, University of Oxford, Oxford, OX3 7BN, UK.
⁵ Department of Internal Medicine, Division of Cardiovascular Medicine, University of Michigan, University of Michigan, Ann Arbor, 48109, MI, USA.
⁶ Department of Human Genetics, University of Michigan, University of Michigan, Ann Arbor, 48109, MI, USA.
⁷ Department of Biostatistics and Center for Statistical Genetics, University of Michigan, Ann Arbor, 48109, MI, USA.
⁸ Department of Clinical Biochemistry, Rigshospitalet, Copenhagen University Hospital, Copenhagen, 2100, Denmark.
⁹ deCODE Genetics/Amgen, Inc., Reykjavik, 101, Iceland.
¹⁰ Department of Clinical Sciences, Malmö, Lund University, Malmö, 221, Sweden.
¹¹ Program in Medical and Population Genetics, Broad Institute, Cambridge, 02142, MA, USA.
¹² MRC Epidemiology Unit, Institute of Metabolic Science, University of Cambridge School of Clinical Medicine, Cambridge Biomedical Campus, Addenbrooke's Hospital, Cambridge, CB2 0QQ, UK.
¹³ Department of Medical Research, Taichung Veterans General Hospital, Taichung, 40705, Taiwan.
¹⁴ Department of Social Work, Tunghai University, Taichung, 40704, Taiwan.
¹⁵ Department of Genetics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, 19104, PA, USA.
¹⁶ Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, 19104, USA.
¹⁷ Division of Cardiology, Department of Medicine; Molecular Physiology Institute, School of Medicine, Duke University, Durham, 27710, NC, USA.
¹⁸ Finnish Institute of Molecular Medicine (FIMM), Helsinki University, Helsinki, 00170, Finland.
¹⁹ Department of Clinical Sciences, Clinical Research Centre, Lund University, Malmö, 221, Sweden.
²⁰ Instituto de Investigaciones Biomédicas, UNAM, Coyoacán, 04510, Mexico City, Mexico.
²¹ Unidad de Biología Molecular y Medicina Genómica, UNAM/INCMNSZ Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, 14080, Mexico.
²² Department of Biostatistics, University of Liverpool, Liverpool, L69 7ZX, UK.
²³ Estonian Genome Center, University of Tartu, Tartu, 50090, Estonia.
²⁴ Institute for Translational Genomics and Population Sciences, Departments of Pediatrics and Medicine, LABioMed at Harbor-UCLA Medical Center, Torrance, 90502, CA, USA.
²⁵ Division of Endocrinology & Metabolism, Department of Internal Medicine, National Taiwan University Hospital, Taipei, 10617, Taiwan.
²⁶ Institute of Preventive Medicine, School of Public Health, National Taiwan University, Taipei, 10617, Taiwan.
²⁷ Department of Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, 19104, PA, USA.
²⁸ Department of Surgery, Corporal Michael Crescenz VA Medical Center, Philadelphia, 19104, PA, USA.
²⁹ Department of Anesthesiology, University of Michigan, Ann Arbor, 48109, MI, USA.
³⁰ Center for Human Genetic Research, Cardiovascular Research Center and Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston, 02114, MA, USA.
³¹ Center for Human Genetic Research, Massachusetts General Hospital, Harvard Medical School, Boston, 02114, MA, USA.
³² Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, 48109, MI, USA.
³³ HUNT Research Centre, Department of Public Health and General Practice, Norwegian University of Science and Technology, Levanger, 7601, Norway.
³⁴ K.G. Jebsen Center for Genetic Epidemiology, Department of Public Health, Norwegian University of Science and Technology, Trondheim, 7491, Norway.
³⁵ Department of Medicine, Levanger Hospital, Nord-Trøndelag Health Trust, Levanger, 7601, Norway.
³⁶ The Copenhagen General Population Study, Herlev and Gentofte Hospital, Copenhagen University Hospital, Copenhagen, 2730, Denmark.
³⁷ Department of Clinical Biochemistry, Herlev and Gentofte Hospital, Copenhagen University Hospital, Copenhagen, 2730, Denmark.
³⁸ The Copenhagen City Heart Study, Frederiksberg Hospital, Copenhagen University Hospital, Copenhagen, 2400, Denmark.
³⁹ Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, 2200, Denmark.
⁴⁰ Department of Molecular Biology, Diabetes Unit, and Center for Human Genetic Research, Massachusetts General Hospital, Boston, 02114, MA, USA.
⁴¹ Departments of Genetics and Medicine, Harvard Medical School, Boston, 02115, MA, USA.
⁴² Department of Biology, Massachusetts Institute of Technology, Cambridge, 02139, MA, USA.
⁴³ Diabetes Unit and Center for Human Genetic Research, Massachusetts General Hospital, Boston, 02115, MA, USA.
⁴⁴ Programs in Metabolism and Medical & Population Genetics, Broad Institute, Cambridge, 02142, MA, USA.
⁴⁵ Department of Medicine, Harvard Medical School, Boston, 02115, MA, USA.
⁴⁶ Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Oxford, OX3 7LE, UK.
⁴⁷ Oxford NIHR Biomedical Research Centre, Churchill Hospital, Oxford, OX4 2PG, UK.
⁴⁸ Regeneron Genetics Center, Tarrytown, 10591, NY, USA. aris.baras@regeneron.com.
⁴⁹ Regeneron Genetics Center, Tarrytown, 10591, NY, USA. rickdewey@gmail.com.
⁵⁰ Regeneron Pharmaceuticals, Tarrytown, 10591, NY, USA. jesper.gromada@regeneron.com.

PMID: 29899519
PMCID: PMC5997992
DOI: 10.1038/s41467-018-04611-z

Genetic inactivation of ANGPTL4 improves glucose homeostasis and is associated with reduced risk of diabetes

Viktoria Gusarova et al. Nat Commun. 2018.

. 2018 Jun 13;9(1):2252.

doi: 10.1038/s41467-018-04611-z.

Authors

Affiliations

¹ Regeneron Pharmaceuticals, Tarrytown, 10591, NY, USA.
² Regeneron Genetics Center, Tarrytown, 10591, NY, USA.
³ Geisinger, Danville, 17822, PA, USA.
⁴ Wellcome Centre for Human Genetics, University of Oxford, Oxford, OX3 7BN, UK.
⁵ Department of Internal Medicine, Division of Cardiovascular Medicine, University of Michigan, University of Michigan, Ann Arbor, 48109, MI, USA.
⁶ Department of Human Genetics, University of Michigan, University of Michigan, Ann Arbor, 48109, MI, USA.
⁷ Department of Biostatistics and Center for Statistical Genetics, University of Michigan, Ann Arbor, 48109, MI, USA.
⁸ Department of Clinical Biochemistry, Rigshospitalet, Copenhagen University Hospital, Copenhagen, 2100, Denmark.
⁹ deCODE Genetics/Amgen, Inc., Reykjavik, 101, Iceland.
¹⁰ Department of Clinical Sciences, Malmö, Lund University, Malmö, 221, Sweden.
¹¹ Program in Medical and Population Genetics, Broad Institute, Cambridge, 02142, MA, USA.
¹² MRC Epidemiology Unit, Institute of Metabolic Science, University of Cambridge School of Clinical Medicine, Cambridge Biomedical Campus, Addenbrooke's Hospital, Cambridge, CB2 0QQ, UK.
¹³ Department of Medical Research, Taichung Veterans General Hospital, Taichung, 40705, Taiwan.
¹⁴ Department of Social Work, Tunghai University, Taichung, 40704, Taiwan.
¹⁵ Department of Genetics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, 19104, PA, USA.
¹⁶ Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, 19104, USA.
¹⁷ Division of Cardiology, Department of Medicine; Molecular Physiology Institute, School of Medicine, Duke University, Durham, 27710, NC, USA.
¹⁸ Finnish Institute of Molecular Medicine (FIMM), Helsinki University, Helsinki, 00170, Finland.
¹⁹ Department of Clinical Sciences, Clinical Research Centre, Lund University, Malmö, 221, Sweden.
²⁰ Instituto de Investigaciones Biomédicas, UNAM, Coyoacán, 04510, Mexico City, Mexico.
²¹ Unidad de Biología Molecular y Medicina Genómica, UNAM/INCMNSZ Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, 14080, Mexico.
²² Department of Biostatistics, University of Liverpool, Liverpool, L69 7ZX, UK.
²³ Estonian Genome Center, University of Tartu, Tartu, 50090, Estonia.
²⁴ Institute for Translational Genomics and Population Sciences, Departments of Pediatrics and Medicine, LABioMed at Harbor-UCLA Medical Center, Torrance, 90502, CA, USA.
²⁵ Division of Endocrinology & Metabolism, Department of Internal Medicine, National Taiwan University Hospital, Taipei, 10617, Taiwan.
²⁶ Institute of Preventive Medicine, School of Public Health, National Taiwan University, Taipei, 10617, Taiwan.
²⁷ Department of Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, 19104, PA, USA.
²⁸ Department of Surgery, Corporal Michael Crescenz VA Medical Center, Philadelphia, 19104, PA, USA.
²⁹ Department of Anesthesiology, University of Michigan, Ann Arbor, 48109, MI, USA.
³⁰ Center for Human Genetic Research, Cardiovascular Research Center and Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston, 02114, MA, USA.
³¹ Center for Human Genetic Research, Massachusetts General Hospital, Harvard Medical School, Boston, 02114, MA, USA.
³² Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, 48109, MI, USA.
³³ HUNT Research Centre, Department of Public Health and General Practice, Norwegian University of Science and Technology, Levanger, 7601, Norway.
³⁴ K.G. Jebsen Center for Genetic Epidemiology, Department of Public Health, Norwegian University of Science and Technology, Trondheim, 7491, Norway.
³⁵ Department of Medicine, Levanger Hospital, Nord-Trøndelag Health Trust, Levanger, 7601, Norway.
³⁶ The Copenhagen General Population Study, Herlev and Gentofte Hospital, Copenhagen University Hospital, Copenhagen, 2730, Denmark.
³⁷ Department of Clinical Biochemistry, Herlev and Gentofte Hospital, Copenhagen University Hospital, Copenhagen, 2730, Denmark.
³⁸ The Copenhagen City Heart Study, Frederiksberg Hospital, Copenhagen University Hospital, Copenhagen, 2400, Denmark.
³⁹ Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, 2200, Denmark.
⁴⁰ Department of Molecular Biology, Diabetes Unit, and Center for Human Genetic Research, Massachusetts General Hospital, Boston, 02114, MA, USA.
⁴¹ Departments of Genetics and Medicine, Harvard Medical School, Boston, 02115, MA, USA.
⁴² Department of Biology, Massachusetts Institute of Technology, Cambridge, 02139, MA, USA.
⁴³ Diabetes Unit and Center for Human Genetic Research, Massachusetts General Hospital, Boston, 02115, MA, USA.
⁴⁴ Programs in Metabolism and Medical & Population Genetics, Broad Institute, Cambridge, 02142, MA, USA.
⁴⁵ Department of Medicine, Harvard Medical School, Boston, 02115, MA, USA.
⁴⁶ Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, Oxford, OX3 7LE, UK.
⁴⁷ Oxford NIHR Biomedical Research Centre, Churchill Hospital, Oxford, OX4 2PG, UK.
⁴⁸ Regeneron Genetics Center, Tarrytown, 10591, NY, USA. aris.baras@regeneron.com.
⁴⁹ Regeneron Genetics Center, Tarrytown, 10591, NY, USA. rickdewey@gmail.com.
⁵⁰ Regeneron Pharmaceuticals, Tarrytown, 10591, NY, USA. jesper.gromada@regeneron.com.

PMID: 29899519
PMCID: PMC5997992
DOI: 10.1038/s41467-018-04611-z

Abstract

Angiopoietin-like 4 (ANGPTL4) is an endogenous inhibitor of lipoprotein lipase that modulates lipid levels, coronary atherosclerosis risk, and nutrient partitioning. We hypothesize that loss of ANGPTL4 function might improve glucose homeostasis and decrease risk of type 2 diabetes (T2D). We investigate protein-altering variants in ANGPTL4 among 58,124 participants in the DiscovEHR human genetics study, with follow-up studies in 82,766 T2D cases and 498,761 controls. Carriers of p.E40K, a variant that abolishes ANGPTL4 ability to inhibit lipoprotein lipase, have lower odds of T2D (odds ratio 0.89, 95% confidence interval 0.85-0.92, p = 6.3 × 10^-10), lower fasting glucose, and greater insulin sensitivity. Predicted loss-of-function variants are associated with lower odds of T2D among 32,015 cases and 84,006 controls (odds ratio 0.71, 95% confidence interval 0.49-0.99, p = 0.041). Functional studies in Angptl4-deficient mice confirm improved insulin sensitivity and glucose homeostasis. In conclusion, genetic inactivation of ANGPTL4 is associated with improved glucose homeostasis and reduced risk of T2D.

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Conflict of interest statement

V.G., C.O.’D., T.M.T., O.G., C.V.V.H., S.B., A.E.L., J.P., A. Marcketta, L.H., A.J.M., I.B.B., J.G.R., J.D.O., G.D.Y., A.R.S., A.B., F.E.D. and J.G. were employees and/or stockholders at Regeneron Pharmaceuticals while engaged in the study. A.B.W. has received consultancy honoraria from Roche. D.F.G., H.H., V.S., P.S., U.T. and K.S. are employed by deCODE Genetics/Amgen Inc.

Figures

**Fig. 1**
*ANGPTL4* p.E40K associates with reduced risk of type 2 diabetes. The association between the p.E40K variant and type 2 diabetes was tested in each study using logistic or Firth logistic regression, coding genotypes according to an additive model. “Combined” effects were calculated using inverse variance weighted fixed-effects meta-analysis. For each study, the squares indicate the odds ratio and lines indicate 95% confidence intervals. The square size is proportional to the standard error of the estimate. CI confidence interval, CGPS Copenhagen General Population Studies, DECODE deCODE, DiscovEHR DiscovEHR Discovery Study, DiscovEHR DiscovEHR-30K, DiscovEHR 30K Replication Study, EINT-C EPIC interact–CoreExome, EINT-Q EPIC Interact–Quad660, ENOR EPIC Norfolk, HUNT the Nord-Trøndelag Health study, MDC Malmo Diet and Cancer Study, MGI the Michigan Genomics Initiative, TD2G/GT2D/DG combined analysis of T2D-GENES, GoT2D, and DIAGRAM studies, UKBB United Kingdom Biobank. The study populations are described in full in Supplementary Table 4 and in the Supplementary Note

**Fig. 2**
Plasma ANGPTL4 levels are reduced in p.G313fs carriers. ANGPTL4 plasma levels were measured in fasted serum from 86 heterozygous p.E40K, 42 heterozygous p.G313fs variant carriers, and 55 controls matched for age, sex, and body mass index. Statistics performed by unpaired t-test with Welch’s correction, comparing each variant carriers group to controls, ****p < 0.0001

**Fig. 3**
K40 and G313fs abolish ANGPTL4 effect on triglycerides. a Plasma triglycerides levels (4 h fasted) in C57Bl/6 mice before (Baseline) and 7 days after (Day 7) hydrodynamic delivery via tail vein injection of cDNA encoding human E40, K40, and Gly313fs ANGPTL4 variants. Control animals were injected with empty vector. b ANGPTL4 plasma levels were measured in the animals described in (a). All groups had five animals. Values are mean ± SEM. Statistics performed by two-way ANOVA with Bonferroni correction, ****p < 0.0001 vs control

**Fig. 4**
*Angptl4*^−/− mice have improved glucose homeostasis. a Serum triglycerides, (b) total cholesterol, and (c) blood glucose levels in *Angptl4*^−/− and littermate control mice on a high-fat diet for 9 weeks. d Oral glucose tolerance test and (e) insulin tolerance test in the animals described in (a–c). All groups had 9–11 animals. Values are mean ± SEM. Statistical analysis by Welch’s t-test (a) and 2-way ANOVA with Sidak’s post-test (d, e), **p < 0.001, ****p < 0.0001. The study was conducted in three different cohorts of mice, with qualitatively similar results in each replicate

**Fig. 5**
*Angptl4*^−/− mice have reduced liver fat. a Body weight, (b) lean and (c) fat mass in *Angptl4*^−/− and littermate control mice on a high-fat diet. d Liver and (e) epididymal white fat weights were evaluated in mice described in (**a–c**) at the time of killing. f Hepatic triglyceride levels and (g) neutral lipid staining in the livers described in (d), scale bar = 200 µm. h Respiratory exchange ratio (RER), (i) locomotor activity, (j) food intake, and (k) energy expenditure were measured during dark and light cycles in *Angptl4*^−/− and littermate control mice on a high-fat diet. All values are mean ± SEM. Statistical analysis was conducted by Welch’s t-test; ^*p < 0.05; ^**p < 0.01. The study was conducted in two different cohorts of mice, with qualitatively similar results in each replicate

See this image and copyright information in PMC

References

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Genetic inactivation of ANGPTL4 improves glucose homeostasis and is associated with reduced risk of diabetes

Affiliations

Genetic inactivation of ANGPTL4 improves glucose homeostasis and is associated with reduced risk of diabetes

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Other Literature Sources

Medical

Molecular Biology Databases