Voltar
Artigo Acesso aberto Revisado por pares
BIBTEX RIS

Role of angiopoietin-like protein 3 in sugar-induced dyslipidemia in rhesus macaques: suppression by fish oil or RNAi

2020; Elsevier BV; Volume: 61; Issue: 3 Linguagem: Inglês

10.1194/jlr.ra119000423

ISSN

1539-7262

Autores

Andrew A. Butler, James L. Graham, Kimber L. Stanhope, So C. Wong, Sarah M. King, Andrew A. Bremer, Ronald M. Krauss, James Hamilton, Peter J. Havel,

Tópico(s)

Liver Disease Diagnosis and Treatment

Resumo

Angiopoietin-like protein 3 (ANGPTL3) inhibits lipid clearance and is a promising target for managing cardiovascular disease. Here we investigated the effects of a high-sugar (high-fructose) diet on circulating ANGPTL3 concentrations in rhesus macaques. Plasma ANGPTL3 concentrations increased ∼30% to 40% after 1 and 3 months of a high-fructose diet (both P < 0.001 vs. baseline). During fructose-induced metabolic dysregulation, plasma ANGPTL3 concentrations were positively correlated with circulating indices of insulin resistance [assessed with fasting insulin and the homeostatic model assessment of insulin resistance (HOMA-IR)], hypertriglyceridemia, adiposity (assessed as leptin), and systemic inflammation [C-reactive peptide (CRP)] and negatively correlated with plasma levels of the insulin-sensitizing hormone adropin. Multiple regression analyses identified a strong association between circulating APOC3 and ANGPTL3 concentrations. Higher baseline plasma levels of both ANGPTL3 and APOC3 were associated with an increased risk for fructose-induced insulin resistance. Fish oil previously shown to prevent insulin resistance and hypertriglyceridemia in this model prevented increases of ANGPTL3 without affecting systemic inflammation (increased plasma CRP and interleukin-6 concentrations). ANGPTL3 RNAi lowered plasma concentrations of ANGPTL3, triglycerides (TGs), VLDL-C, APOC3, and APOE. These decreases were consistent with a reduced risk of atherosclerosis. In summary, dietary sugar-induced increases of circulating ANGPTL3 concentrations after metabolic dysregulation correlated positively with leptin levels, HOMA-IR, and dyslipidemia. Targeting ANGPTL3 expression with RNAi inhibited dyslipidemia by lowering plasma TGs, VLDL-C, APOC3, and APOE levels in rhesus macaques. Angiopoietin-like protein 3 (ANGPTL3) inhibits lipid clearance and is a promising target for managing cardiovascular disease. Here we investigated the effects of a high-sugar (high-fructose) diet on circulating ANGPTL3 concentrations in rhesus macaques. Plasma ANGPTL3 concentrations increased ∼30% to 40% after 1 and 3 months of a high-fructose diet (both P < 0.001 vs. baseline). During fructose-induced metabolic dysregulation, plasma ANGPTL3 concentrations were positively correlated with circulating indices of insulin resistance [assessed with fasting insulin and the homeostatic model assessment of insulin resistance (HOMA-IR)], hypertriglyceridemia, adiposity (assessed as leptin), and systemic inflammation [C-reactive peptide (CRP)] and negatively correlated with plasma levels of the insulin-sensitizing hormone adropin. Multiple regression analyses identified a strong association between circulating APOC3 and ANGPTL3 concentrations. Higher baseline plasma levels of both ANGPTL3 and APOC3 were associated with an increased risk for fructose-induced insulin resistance. Fish oil previously shown to prevent insulin resistance and hypertriglyceridemia in this model prevented increases of ANGPTL3 without affecting systemic inflammation (increased plasma CRP and interleukin-6 concentrations). ANGPTL3 RNAi lowered plasma concentrations of ANGPTL3, triglycerides (TGs), VLDL-C, APOC3, and APOE. These decreases were consistent with a reduced risk of atherosclerosis. In summary, dietary sugar-induced increases of circulating ANGPTL3 concentrations after metabolic dysregulation correlated positively with leptin levels, HOMA-IR, and dyslipidemia. Targeting ANGPTL3 expression with RNAi inhibited dyslipidemia by lowering plasma TGs, VLDL-C, APOC3, and APOE levels in rhesus macaques. Elevated circulating lipids and lipoproteins are the major known modifiable risk factors for CVD, the leading cause of death in the United States (1Chait A. Subramanian S. Hypertriglyceridemia: pathophysiology, role of genetics, consequences, and treatment.in: Feingold K.R. Anawalt B. Boyce A. Endotext [Internet]. MDText.com, Inc., South Dartmouth, MA2000Google Scholar, 2Baigent C. Blackwell L. Emberson J. Holland L.E. Reith C. Bhala N. Peto R. Barnes E.H. Keech A. Simes J. et al.Efficacy and safety of more intensive lowering of LDL cholesterol: a meta-analysis of data from 170,000 participants in 26 randomised trials.Lancet. 2010; 376: 1670-1681Abstract Full Text Full Text PDF PubMed Scopus (4534) Google Scholar, 3Navarese E.P. Robinson J.G. Kowalewski M. Kolodziejczak M. Andreotti F. Bliden K. Tantry U. Kubica J. Raggi P. Gurbel P.A. Association between baseline LDL-C level and total and cardiovascular mortality after LDL-C lowering: a systematic review and meta-analysis.JAMA. 2018; 319: 1566-1579Crossref PubMed Scopus (255) Google Scholar, 4Siri-Tarino P.W. Krauss R.M. The early years of lipoprotein research: from discovery to clinical application.J. Lipid Res. 2016; 57: 1771-1777Abstract Full Text Full Text PDF PubMed Scopus (19) Google Scholar). Severe hypertriglyceridemia (e.g., >800 mg/dl) also increases the risk for acute pancreatitis, which can be life-threatening (5Hegele R.A. Ginsberg H.N. Chapman M.J. Nordestgaard B.G. Kuivenhoven J.A. Averna M. Boren J. Bruckert E. Catapano A.L. Descamps O.S. et al.The polygenic nature of hypertriglyceridaemia: implications for definition, diagnosis, and management.Lancet Diabetes Endocrinol. 2014; 2: 655-666Abstract Full Text Full Text PDF PubMed Scopus (404) Google Scholar). The prevalence of hyperlipidemias increases with obesity and aging, although less common genetic disorders can also lead to moderate to severe dyslipidemias. Current treatments focus on reducing fasting plasma concentrations of cholesterol packaged in LDLs and triglycerides (TGs). Multiple treatment options are available that lower plasma lipids/lipoproteins. However, monogenic disorders causing familial hypercholesterolemia or hypertriglyceridemia and polygenic disorders causing severe hypertriglyceridemia often require alternative approaches to achieve treatment goals (1Chait A. Subramanian S. Hypertriglyceridemia: pathophysiology, role of genetics, consequences, and treatment.in: Feingold K.R. Anawalt B. Boyce A. Endotext [Internet]. MDText.com, Inc., South Dartmouth, MA2000Google Scholar, 6Berberich A.J. Hegele R.A. The complex molecular genetics of familial hypercholesterolaemia.Nat. Rev. Cardiol. 2019; 16: 9-20Crossref PubMed Scopus (132) Google Scholar). New approaches for increasing lipid clearance from the circulation and reducing residual risk for CVD are needed. Angiopoietin-like protein 3 (ANGPTL3) is a secretory protein expressed in the liver (a "hepatokine") and is considered a promising lead target in the development of lipid-lowering therapies (7Nordestgaard B.G. Nicholls S.J. Langsted A. Ray K.K. Tybjaerg-Hansen A. Advances in lipid-lowering therapy through gene-silencing technologies.Nat. Rev. Cardiol. 2018; 15: 261-272Crossref PubMed Scopus (73) Google Scholar). ANGPTL3 belongs to a family of secretory proteins (ANGPTL3, ANGPTL4, and ANGPTL8) that affect the clearance of circulating lipids by regulating endothelial lipase and lipoprotein lipase (8Kersten S. Angiopoietin-like 3 in lipoprotein metabolism.Nat. Rev. Endocrinol. 2017; 13: 731-739Crossref PubMed Scopus (107) Google Scholar). Specifically, ANGPTL3 interacts with ANGPTL8 to suppress lipoprotein lipase activity after feeding (9Quagliarini F. Wang Y. Kozlitina J. Grishin N.V. Hyde R. Boerwinkle E. Valenzuela D.M. Murphy A.J. Cohen J.C. Hobbs H.H. Atypical angiopoietin-like protein that regulates ANGPTL3.Proc. Natl. Acad. Sci. USA. 2012; 109: 19751-19756Crossref PubMed Scopus (341) Google Scholar, 10Haller J.F. Mintah I.J. Shihanian L.M. Stevis P. Buckler D. Alexa-Braun C.A. Kleiner S. Banfi S. Cohen J.C. Hobbs H.H. et al.ANGPTL8 requires ANGPTL3 to inhibit lipoprotein lipase and plasma triglyceride clearance.J. Lipid Res. 2017; 58: 1166-1173Abstract Full Text Full Text PDF PubMed Scopus (122) Google Scholar, 11Chi X. Britt E.C. Shows H.W. Hjelmaas A.J. Shetty S.K. Cushing E.M. Li W. Dou A. Zhang R. Davies B.S.J. ANGPTL8 promotes the ability of ANGPTL3 to bind and inhibit lipoprotein lipase.Mol. Metab. 2017; 6: 1137-1149Crossref PubMed Scopus (109) Google Scholar). ANGPTL3 also regulates the clearance of HDLs by binding to and inhibiting the activity of endothelial lipase (12Shimamura M. Matsuda M. Yasumo H. Okazaki M. Fujimoto K. Kono K. Shimizugawa T. Ando Y. Koishi R. Kohama T. et al.Angiopoietin-like protein3 regulates plasma HDL cholesterol through suppression of endothelial lipase.Arterioscler. Thromb. Vasc. Biol. 2007; 27: 366-372Crossref PubMed Scopus (211) Google Scholar). The clinical potential of ANGPTL3 for reducing plasma lipids is supported by human genetics. Specifically, inactivating ANGPTL3 gene variants associate with lower plasma levels of LDL-C and TGs and a lower risk of CVD (13Musunuru K. Pirruccello J.P. Do R. Peloso G.M. Guiducci C. Sougnez C. Garimella K.V. Fisher S. Abreu J. Barry A.J. et al.Exome sequencing, ANGPTL3 mutations, and familial combined hypolipidemia.N. Engl. J. Med. 2010; 363: 2220-2227Crossref PubMed Scopus (503) Google Scholar, 14Minicocci I. Montali A. Robciuc M.R. Quagliarini F. Censi V. Labbadia G. Gabiati C. Pigna G. Sepe M.L. Pannozzo F. et al.Mutations in the ANGPTL3 gene and familial combined hypolipidemia: a clinical and biochemical characterization.J. Clin. Endocrinol. Metab. 2012; 97: E1266-E1275Crossref PubMed Scopus (99) Google Scholar, 15Robciuc M.R. Maranghi M. Lahikainen A. Rader D. Bensadoun A. Oorni K. Metso J. Minicocci I. Ciociola E. Ceci F. et al.Angptl3 deficiency is associated with increased insulin sensitivity, lipoprotein lipase activity, and decreased serum free fatty acids.Arterioscler. Thromb. Vasc. Biol. 2013; 33 (–. [Erratum. . Arterioscler. Thromb. Vasc. Biol. 33 2013 1706–1713 e124): 1706-1713Crossref PubMed Scopus (122) Google Scholar, 16Pisciotta L. Favari E. Magnolo L. Simonelli S. Adorni M.P. Sallo R. Fancello T. Zavaroni I. Ardigo D. Bernini F. et al.Characterization of three kindreds with familial combined hypolipidemia caused by loss-of-function mutations of ANGPTL3.Circ Cardiovasc Genet. 2012; 5: 42-50Crossref PubMed Scopus (100) Google Scholar, 17Martín-Campos J.M. Roig R. Mayoral C. Martinez S. Marti G. Arroyo J.A. Julve J. Blanco-Vaca F. Identification of a novel mutation in the ANGPTL3 gene in two families diagnosed of familial hypobetalipoproteinemia without APOB mutation.Clin. Chim. Acta. 2012; 413: 552-555Crossref PubMed Scopus (50) Google Scholar, 18Noto D. Cefalu A.B. Valenti V. Fayer F. Pinotti E. Ditta M. Spina R. Vigna G. Yue P. Kathiresan S. et al.Prevalence of ANGPTL3 and APOB gene mutations in subjects with combined hypolipidemia.Arterioscler. Thromb. Vasc. Biol. 2012; 32: 805-809Crossref PubMed Scopus (65) Google Scholar, 19Minicocci I. Santini S. Cantisani V. Stitziel N. Kathiresan S. Arroyo J.A. Marti G. Pisciotta L. Noto D. Cefalu A.B. et al.Clinical characteristics and plasma lipids in subjects with familial combined hypolipidemia: a pooled analysis.J. Lipid Res. 2013; 54: 3481-3490Abstract Full Text Full Text PDF PubMed Scopus (59) Google Scholar). Moreover, small trials using antisense or monoclonal antibodies to inhibit ANGPTL3 have shown clinically significant reductions in plasma LDL-C and TG concentrations in patients with familial hypercholesterolemia (20Gaudet D. Gipe D.A. Pordy R. Ahmad Z. Cuchel M. Shah P.K. Chyu K.Y. Sasiela W.J. Chan K.C. Brisson D. et al.ANGPTL3 inhibition in homozygous familial hypercholesterolemia.N. Engl. J. Med. 2017; 377: 296-297Crossref PubMed Scopus (201) Google Scholar, 21Dewey F.E. Gusarova V. Dunbar R.L. O'Dushlaine C. Schurmann C. Gottesman O. McCarthy S. Van Hout C.V. Bruse S. Dansky H.M. et al.Genetic and pharmacologic inactivation of ANGPTL3 and cardiovascular disease.N. Engl. J. Med. 2017; 377: 211-221Crossref PubMed Scopus (462) Google Scholar, 22Graham M.J. Lee R.G. Brandt T.A. Tai L.J. Fu W. Peralta R. Yu R. Hurh E. Paz E. McEvoy B.W. et al.Cardiovascular and metabolic effects of ANGPTL3 antisense oligonucleotides.N. Engl. J. Med. 2017; 377: 222-232Crossref PubMed Scopus (384) Google Scholar). APOC3 has been recently targeted for the treatment of dyslipidemia. APOC3 is a 8.8 kDa glycoprotein released primarily from the liver and is a major structural component of atherogenic VLDL particles; it is also found in HDLs and chylomicron particles (23Ramms B. Gordts P. Apolipoprotein C-III in triglyceride-rich lipoprotein metabolism.Curr. Opin. Lipidol. 2018; 29: 171-179Crossref PubMed Scopus (60) Google Scholar). APOC3 inhibits the processing of large, TG-rich VLDLs (23Ramms B. Gordts P. Apolipoprotein C-III in triglyceride-rich lipoprotein metabolism.Curr. Opin. Lipidol. 2018; 29: 171-179Crossref PubMed Scopus (60) Google Scholar) and has important intracellular functions in hepatocytes that facilitate VLDL production (24Sundaram M. Zhong S. Bou Khalil M. Links P.H. Zhao Y. Iqbal J. Hussain M.M. Parks R.J. Wang Y. Yao Z. Expression of apolipoprotein C-III in McA-RH7777 cells enhances VLDL assembly and secretion under lipid-rich conditions.J. Lipid Res. 2010; 51: 150-161Abstract Full Text Full Text PDF PubMed Scopus (122) Google Scholar). Loss-of-function APOC3 gene variants associate with significant (∼40%) reductions in CVD risk and plasma concentrations of TGs and LDL-C (6Berberich A.J. Hegele R.A. The complex molecular genetics of familial hypercholesterolaemia.Nat. Rev. Cardiol. 2019; 16: 9-20Crossref PubMed Scopus (132) Google Scholar, 7Nordestgaard B.G. Nicholls S.J. Langsted A. Ray K.K. Tybjaerg-Hansen A. Advances in lipid-lowering therapy through gene-silencing technologies.Nat. Rev. Cardiol. 2018; 15: 261-272Crossref PubMed Scopus (73) Google Scholar, 8Kersten S. Angiopoietin-like 3 in lipoprotein metabolism.Nat. Rev. Endocrinol. 2017; 13: 731-739Crossref PubMed Scopus (107) Google Scholar, 9Quagliarini F. Wang Y. Kozlitina J. Grishin N.V. Hyde R. Boerwinkle E. Valenzuela D.M. Murphy A.J. Cohen J.C. Hobbs H.H. Atypical angiopoietin-like protein that regulates ANGPTL3.Proc. Natl. Acad. Sci. USA. 2012; 109: 19751-19756Crossref PubMed Scopus (341) Google Scholar). Preclinical studies in rodent and porcine models indicate that increased APOC3 synthesis is sufficient for producing hypertriglyceridemia (10Haller J.F. Mintah I.J. Shihanian L.M. Stevis P. Buckler D. Alexa-Braun C.A. Kleiner S. Banfi S. Cohen J.C. Hobbs H.H. et al.ANGPTL8 requires ANGPTL3 to inhibit lipoprotein lipase and plasma triglyceride clearance.J. Lipid Res. 2017; 58: 1166-1173Abstract Full Text Full Text PDF PubMed Scopus (122) Google Scholar, 11Chi X. Britt E.C. Shows H.W. Hjelmaas A.J. Shetty S.K. Cushing E.M. Li W. Dou A. Zhang R. Davies B.S.J. ANGPTL8 promotes the ability of ANGPTL3 to bind and inhibit lipoprotein lipase.Mol. Metab. 2017; 6: 1137-1149Crossref PubMed Scopus (109) Google Scholar, 25Wei J. Ouyang H. Wang Y. Pang D. Cong N.X. Wang T. Leng B. Li D. Li X. Wu R. et al.Characterization of a hypertriglyceridemic transgenic miniature pig model expressing human apolipoprotein CIII.FEBS J. 2012; 279: 91-99Crossref PubMed Scopus (48) Google Scholar). Clinical trials using antisense oligonucleotides to suppress APOC3 synthesis (Volanesorsen, Ionis 304801) have demonstrated substantial antihyperlipidemic effects in patients with severe hypertriglyceridemia (12Shimamura M. Matsuda M. Yasumo H. Okazaki M. Fujimoto K. Kono K. Shimizugawa T. Ando Y. Koishi R. Kohama T. et al.Angiopoietin-like protein3 regulates plasma HDL cholesterol through suppression of endothelial lipase.Arterioscler. Thromb. Vasc. Biol. 2007; 27: 366-372Crossref PubMed Scopus (211) Google Scholar, 13Musunuru K. Pirruccello J.P. Do R. Peloso G.M. Guiducci C. Sougnez C. Garimella K.V. Fisher S. Abreu J. Barry A.J. et al.Exome sequencing, ANGPTL3 mutations, and familial combined hypolipidemia.N. Engl. J. Med. 2010; 363: 2220-2227Crossref PubMed Scopus (503) Google Scholar). Nonhuman primates are an important resource for the translational research of novel therapies against obesity-related metabolic diseases (26Havel P.J. Kievit P. Comuzzie A.G. Bremer A.A. Use and importance of nonhuman primates in metabolic disease research: current state of the field.ILAR J. 2017; 58: 251-268Crossref PubMed Scopus (35) Google Scholar, 27Cox L.A. Olivier M. Spradling-Reeves K. Karere G.M. Comuzzie A.G. VandeBerg J.L. Nonhuman primates and translational research-cardiovascular disease.ILAR J. 2017; 58: 235-250Crossref PubMed Scopus (35) Google Scholar). Previous studies have demonstrated that rhesus macaques provided with fructose or high-fructose corn syrup (HFCS) supplements (300–600 kcal/d) rapidly gain weight and develop features of metabolic syndrome (insulin resistance, hypertriglyceridemia, and increased APOC3) (28Bremer A.A. Stanhope K.L. Graham J.L. Cummings B.P. Wang W. Saville B.R. Havel P.J. Fructose-fed rhesus monkeys: a nonhuman primate model of insulin resistance, metabolic syndrome, and type 2 diabetes.Clin. Transl. Sci. 2011; 4: 243-252Crossref PubMed Scopus (126) Google Scholar, 29Bremer A.A. Stanhope K.L. Graham J.L. Cummings B.P. Ampah S.B. Saville B.R. Havel P.J. Fish oil supplementation ameliorates fructose-induced hypertriglyceridemia and insulin resistance in adult male rhesus macaques.J. Nutr. 2014; 144: 5-11Crossref PubMed Scopus (51) Google Scholar, 30Butler A.A. Price C.A. Graham J.L. Stanhope K.L. King S. Hung Y.H. Sethupathy P. Wong S. Hamilton J. Krauss R.M. et al.Fructose-induced hypertriglyceridemia in rhesus macaques is attenuated with fish oil or ApoC3 RNA interference.J. Lipid Res. 2019; 60: 805-818Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar). Using this model, we recently reported that an RNAi construct targeting hepatic APOC3 expression reduces plasma TG concentrations (30Butler A.A. Price C.A. Graham J.L. Stanhope K.L. King S. Hung Y.H. Sethupathy P. Wong S. Hamilton J. Krauss R.M. et al.Fructose-induced hypertriglyceridemia in rhesus macaques is attenuated with fish oil or ApoC3 RNA interference.J. Lipid Res. 2019; 60: 805-818Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar). Moreover, elevations of plasma TG concentrations during fructose consumption are positively correlated with increases of circulating APOC3 concentrations (30Butler A.A. Price C.A. Graham J.L. Stanhope K.L. King S. Hung Y.H. Sethupathy P. Wong S. Hamilton J. Krauss R.M. et al.Fructose-induced hypertriglyceridemia in rhesus macaques is attenuated with fish oil or ApoC3 RNA interference.J. Lipid Res. 2019; 60: 805-818Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar). However, the impact of dietary composition and diet-induced obesity/metabolic dysfunction on plasma ANGPTL3 concentrations has not been investigated. The current experiments determined the effects of a high-sugar diet on circulating ANGPTL3 levels and the effects of fish oil supplementation in male rhesus macaques. The results of a large (n = 59) study into the effects of fructose supplements on weight gain and indices of insulin resistance [fasting insulin, fasting glucose, and homeostatic model assessment of insulin resistance (HOMA-IR)] and dyslipidemia have previously been reported (28Bremer A.A. Stanhope K.L. Graham J.L. Cummings B.P. Wang W. Saville B.R. Havel P.J. Fructose-fed rhesus monkeys: a nonhuman primate model of insulin resistance, metabolic syndrome, and type 2 diabetes.Clin. Transl. Sci. 2011; 4: 243-252Crossref PubMed Scopus (126) Google Scholar, 29Bremer A.A. Stanhope K.L. Graham J.L. Cummings B.P. Ampah S.B. Saville B.R. Havel P.J. Fish oil supplementation ameliorates fructose-induced hypertriglyceridemia and insulin resistance in adult male rhesus macaques.J. Nutr. 2014; 144: 5-11Crossref PubMed Scopus (51) Google Scholar, 30Butler A.A. Price C.A. Graham J.L. Stanhope K.L. King S. Hung Y.H. Sethupathy P. Wong S. Hamilton J. Krauss R.M. et al.Fructose-induced hypertriglyceridemia in rhesus macaques is attenuated with fish oil or ApoC3 RNA interference.J. Lipid Res. 2019; 60: 805-818Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar, 31Butler A.A. Zhang J. Price C.A. Stevens J.R. Graham J.L. Stanhope K.L. King S. Krauss R.M. Bremer A.A. Havel P.J. Low plasma adropin concentrations increase risks of weight gain and metabolic dysregulation in response to a high-sugar diet in male nonhuman primates.J. Biol. Chem. 2019; 294: 9706-9719Abstract Full Text Full Text PDF PubMed Scopus (32) Google Scholar). These studies reported rapid gains in body weight, fasting hyperinsulinemia, fasting hypertriglyceridemia, and elevated plasma APOC3 concentrations with fructose consumption that are largely prevented by fish oil supplementation (29Bremer A.A. Stanhope K.L. Graham J.L. Cummings B.P. Ampah S.B. Saville B.R. Havel P.J. Fish oil supplementation ameliorates fructose-induced hypertriglyceridemia and insulin resistance in adult male rhesus macaques.J. Nutr. 2014; 144: 5-11Crossref PubMed Scopus (51) Google Scholar, 30Butler A.A. Price C.A. Graham J.L. Stanhope K.L. King S. Hung Y.H. Sethupathy P. Wong S. Hamilton J. Krauss R.M. et al.Fructose-induced hypertriglyceridemia in rhesus macaques is attenuated with fish oil or ApoC3 RNA interference.J. Lipid Res. 2019; 60: 805-818Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar). The current investigation examined dietary effects and relationships between plasma concentrations of ANGPTL3 and indices of insulin resistance, systemic inflammation, and lipoprotein metabolism. We also report on the effects of dietary fish oil supplementation on ANGPTL3 responses to fructose. The plasma samples used for this experiment are from a study whose outcomes were reported previously (29Bremer A.A. Stanhope K.L. Graham J.L. Cummings B.P. Ampah S.B. Saville B.R. Havel P.J. Fish oil supplementation ameliorates fructose-induced hypertriglyceridemia and insulin resistance in adult male rhesus macaques.J. Nutr. 2014; 144: 5-11Crossref PubMed Scopus (51) Google Scholar). Finally, we report on the effects of inhibiting hepatic ANGPTL3 expression with RNAi on plasma lipid/lipoprotein profiles. This study includes comparisons with the responses to inhibiting hepatic APOC3 concentrations, which have been reported previously (30Butler A.A. Price C.A. Graham J.L. Stanhope K.L. King S. Hung Y.H. Sethupathy P. Wong S. Hamilton J. Krauss R.M. et al.Fructose-induced hypertriglyceridemia in rhesus macaques is attenuated with fish oil or ApoC3 RNA interference.J. Lipid Res. 2019; 60: 805-818Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar). Protocols for all animal studies were approved by the University of California, Davis Institutional Animal Care and Use Committee and were conducted in accordance with the U.S. Department of Agriculture Animal Welfare Act and the National Institutes of Health's Guide for the Care and Use of Laboratory Animals. The animal studies, feeding protocols, and dietary effects of fructose and fish oil on body weight and blood chemistries have been described previously (29Bremer A.A. Stanhope K.L. Graham J.L. Cummings B.P. Ampah S.B. Saville B.R. Havel P.J. Fish oil supplementation ameliorates fructose-induced hypertriglyceridemia and insulin resistance in adult male rhesus macaques.J. Nutr. 2014; 144: 5-11Crossref PubMed Scopus (51) Google Scholar, 30Butler A.A. Price C.A. Graham J.L. Stanhope K.L. King S. Hung Y.H. Sethupathy P. Wong S. Hamilton J. Krauss R.M. et al.Fructose-induced hypertriglyceridemia in rhesus macaques is attenuated with fish oil or ApoC3 RNA interference.J. Lipid Res. 2019; 60: 805-818Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar, 31Butler A.A. Zhang J. Price C.A. Stevens J.R. Graham J.L. Stanhope K.L. King S. Krauss R.M. Bremer A.A. Havel P.J. Low plasma adropin concentrations increase risks of weight gain and metabolic dysregulation in response to a high-sugar diet in male nonhuman primates.J. Biol. Chem. 2019; 294: 9706-9719Abstract Full Text Full Text PDF PubMed Scopus (32) Google Scholar). In brief, adult male rhesus macaques (n = 59) (age: 12.0 ± 2.8 years; range: 6.4–17.8 years) maintained at the California National Primate Research Center were provided a standard commercial nonhuman primate diet (5047; LabDiet, St. Louis, MO) and water. This grain-based diet provides 30%/kcal as protein, 11%/kcal as fat, and 59%/kcal as complex carbohydrates. After determining baseline body weights and collecting fasting blood samples, animals were provided a solution containing 75 g fructose (300 kcal) daily in a total volume of 500 ml flavored Kool-Aid (Kraft Foods, Chicago, IL) beverages. Male rhesus macaques consume on average 800–900 kcal/day. These animals consumed approximately 30% of their daily caloric intake from fructose. Body weight measurements and fasting blood samples were collected after 1 and 3 months of fructose consumption. Ten additional adult male fructose-fed rhesus monkeys were supplemented with 4 g/day whole fish oil (Jedwards International, Inc., Braintree, MA) for 6 months. These animals were compared with a subset of nine animals from the group of 59 animals that were studied concurrently for 6 months (29Bremer A.A. Stanhope K.L. Graham J.L. Cummings B.P. Ampah S.B. Saville B.R. Havel P.J. Fish oil supplementation ameliorates fructose-induced hypertriglyceridemia and insulin resistance in adult male rhesus macaques.J. Nutr. 2014; 144: 5-11Crossref PubMed Scopus (51) Google Scholar). Body weight measurements and fasting blood samples from these animals were collected at baseline and after 1, 3, and 6 months. The six animals used for this experiment received a modified moderate-fat diet protocol supplemented with HFCS. The rationale for this protocol was to achieve a closer match to a typical human diet and to maximize hypertriglyceridemia. The animals were provided HFCS-sweetened beverages (500 ml; 15% by weight sugar) twice per day, ingesting a total of 150 g (2 servings/day of 300 kcal for a total of 600 kcal/d) of HFCS. HFCS is 55% fructose (330 kcal) and 45% glucose (270 kcal). These animals are likely to have received up to 60% of energy intake in the form of simple sugars. The New World monkey diet (LabDiet) has moderate fat content (23%/kcal vs. 11%/kcal as fat for the 5047 diet). Fasting blood samples were collected at baseline, after which animals were started on the HFCS/moderate fat diet. After 6 weeks, an RNAi construct targeting ANGPTL3 mRNA (ARO-ANGPTL3; 4 mg/kg) was administered in four animals via subcutaneous injection on day 0 and day 29 (n = 4). Two additional animals received vehicle injections. ARO-ANGPTL3 is a synthetic double-stranded (both strands contain 21 nucleotides) hepatocyte-targeted N-acetylgalactosamine-conjugated RNAi molecule. The N-acetylgalactosamine moiety targets the RNAi into hepatocytes by acting as a ligand for the highly expressed hepatocyte-specific asialoglycoprotein receptor. The RNAi construct was designed to silence ANGPTL3 mRNA in hepatocytes in humans and nonhuman primates with high specificity. The RNAi was synthesized with 2′-O-methyl/2′-fluoro-modified nucleotides in order to be resistant to nucleases and to abrogate potential immune activation (32Robbins M. Judge A. MacLachlan I. siRNA and innate immunity.Oligonucleotides. 2009; 19: 89-102Crossref PubMed Scopus (321) Google Scholar). Fasting blood samples were collected at days 8, 15, 21, 29, 36, 43, 50, 57, 71, and 85. Plasma glucose concentrations were measured using a glucose analyzer (YSI Life Sciences, Yellow Springs, OH). Plasma adiponectin, insulin, and leptin concentrations were measured using RIA assays from Millipore (Burlington, MA). Plasma total cholesterol (TC), HDL-C, direct LDL-C, TGs, APOA1, APOB, APOC3, and APOE concentrations were measured using a Polychem chemistry analyzer (Polymedco, Cortlandt Manor, NY); reagents were purchased from MedTest DX (Canton, MI). VLDL-C was calculated by subtracting HDL-C and LDL-C from TC. Plasma ANGPTL3 concentrations were measured using ELISA (DANL30; R&D Systems, Minneapolis, MN) for human ANGPTL3 that cross-reacts with macaque ANGPTL3. Plasma adropin concentrations were measured by ELISA (Peninsula Laboratories, San Carlos, CA). Plasma concentrations of VLDL and IDL, LDL, and HDL particle subfractions were measured using specific particle-size intervals determined by ion mobility. This approach allows for direct particle quantification as a function of particle diameter (33Caulfield M.P. Li S. Lee G. Blanche P.J. Salameh W.A. Benner W.H. Reitz R.E. Krauss R.M. Direct determination of lipoprotein particle sizes and concentrations by ion mobility analysis.Clin. Chem. 2008; 54: 1307-1316Crossref PubMed Scopus (173) Google Scholar), following a procedure to remove other plasma proteins (34Mora S. Caulfield M.P. Wohlgemuth J. Chen Z. Superko H.R. Rowland C.M. Glynn R.J. Ridker P.M. Krauss R.M. Atherogenic lipoprotein subfractions determined by ion mobility and first cardiovascular events after random allocation to high-intensity statin or placebo: the Justification for the Use of Statins in Prevention: an Intervention Trial Evaluating Rosuvastatin (JUPITER) Trial.Circulation. 2015; 132: 2220-2229Crossref PubMed Scopus (82) Google Scholar). The ion mobility instrument uses an electrospray to create an aerosol of particles that then pass through a differential mobility analyzer coupled to a particle counter. Particle concentrations (in nanomoles per liter) are determined for subfractions defined by the following size intervals: VLDL: large (42.40–54.70 nm), medium (33.50–42.39 nm), and small (29.60–33.49 nm); IDL: large (25.00–29.59 nm) and small (23.33–24.99 nm); LDL: large (22.0–23.32 nm), medium (21.41–21.99 nm), small (20.82–21.40 nm), and very small (18.0–20.81 nm); and HDL: large (10.50–14.50 nm) and small (7.65–10.49 nm). Peak LDL diameter (in nanometers) was determined as previously described (33Caulfield M.P. Li S. Lee G. Blanche P.J. Salameh W.A. Benner W.H. Reitz R.E

Referência(s)