Inhibition of miR-33a/b in non-human primates raises plasma HDL and lowers VLDL triglycerides (original) (raw)

Nature volume 478, pages 404–407 (2011)Cite this article

Subjects

Abstract

Cardiovascular disease remains the leading cause of mortality in westernized countries, despite optimum medical therapy to reduce the levels of low-density lipoprotein (LDL)-associated cholesterol. The pursuit of novel therapies to target the residual risk has focused on raising the levels of high-density lipoprotein (HDL)-associated cholesterol in order to exploit its atheroprotective effects1. MicroRNAs (miRNAs) have emerged as important post-transcriptional regulators of lipid metabolism and are thus a new class of target for therapeutic intervention2. MicroRNA-33a and microRNA-33b (miR-33a/b) are intronic miRNAs whose encoding regions are embedded in the sterol-response-element-binding protein genes SREBF2 and SREBF1 (refs 3–5), respectively. These miRNAs repress expression of the cholesterol transporter ABCA1, which is a key regulator of HDL biogenesis. Recent studies in mice suggest that antagonizing miR-33a may be an effective strategy for raising plasma HDL levels3,4,5 and providing protection against atherosclerosis6; however, extrapolating these findings to humans is complicated by the fact that mice lack miR-33b, which is present only in the SREBF1 gene of medium and large mammals. Here we show in African green monkeys that systemic delivery of an anti-miRNA oligonucleotide that targets both miR-33a and miR-33b increased hepatic expression of ABCA1 and induced a sustained increase in plasma HDL levels over 12 weeks. Notably, miR-33 antagonism in this non-human primate model also increased the expression of miR-33 target genes involved in fatty acid oxidation (CROT, CPT1A, HADHB and PRKAA1) and reduced the expression of genes involved in fatty acid synthesis (SREBF1, FASN, ACLY and ACACA), resulting in a marked suppression of the plasma levels of very-low-density lipoprotein (VLDL)-associated triglycerides, a finding that has not previously been observed in mice. These data establish, in a model that is highly relevant to humans, that pharmacological inhibition of miR-33a and miR-33b is a promising therapeutic strategy to raise plasma HDL and lower VLDL triglyceride levels for the treatment of dyslipidaemias that increase cardiovascular disease risk.

This is a preview of subscription content, access via your institution

Access options

Subscribe to this journal

Receive 51 print issues and online access

$199.00 per year

only $3.90 per issue

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

Similar content being viewed by others

Accession codes

Data deposits

The microarray data have been deposited in the Gene Expression Omnibus database under accession number GSE31177.

References

  1. deGoma, E. M. & Rader, D. J. Novel HDL-directed pharmacotherapeutic strategies. Nature Rev. Cardiol. 8, 266–277 (2011)
    Article CAS Google Scholar
  2. Moore, K. J., Rayner, K. J., Suarez, Y. & Fernandez-Hernando, C. MicroRNAs and cholesterol metabolism. Trends Endocrinol. Metab. 21, 699–706 (2010)
    Article CAS Google Scholar
  3. Marquart, T. J., Allen, R. M., Ory, D. S. & Baldan, A. miR-33 links SREBP-2 induction to repression of sterol transporters. Proc. Natl Acad. Sci. USA 107, 12228–12232 (2010)
    Article ADS CAS Google Scholar
  4. Najafi-Shoushtari, S. H. et al. MicroRNA-33 and the SREBP host genes cooperate to control cholesterol homeostasis. Science 328, 1566–1569 (2010)
    Article ADS CAS Google Scholar
  5. Rayner, K. J. et al. miR-33 contributes to the regulation of cholesterol homeostasis. Science 328, 1570–1573 (2010)
    Article ADS CAS Google Scholar
  6. Rayner, K. J. et al. Antagonism of miR-33 in mice promotes reverse cholesterol transport and regression of atherosclerosis. J. Clin. Invest. 121, 2921–2931 (2011)
    Article CAS Google Scholar
  7. Davalos, A. et al. miR-33a/b contribute to the regulation of fatty acid metabolism and insulin signaling. Proc. Natl Acad. Sci. USA 108, 9232–9237 (2011)
    Article ADS CAS Google Scholar
  8. Gerin, I. et al. Expression of miR-33 from an SREBP2 intron inhibits cholesterol export and fatty acid oxidation. J. Biol. Chem. 285, 33652–33661 (2010)
    Article CAS Google Scholar
  9. Horton, J. D., Goldstein, J. L. & Brown, M. S. SREBPs: activators of the complete program of cholesterol and fatty acid synthesis in the liver. J. Clin. Invest. 109, 1125–1131 (2002)
    Article CAS Google Scholar
  10. Horie, T. et al. microRNA-33 encoded by an intron of sterol regulatory element-binding protein 2 (Srebp2) regulates HDL in vivo . Proc. Natl Acad. Sci. USA 107, 17321–17326 (2010)
    Article ADS CAS Google Scholar
  11. Geary, R. S. Antisense oligonucleotide pharmacokinetics and metabolism. Expert Opin. Drug Metab. Toxicol. 5, 381–391 (2009)
    Article CAS Google Scholar
  12. Chyu, K. Y., Peter, A. & Shah, P. K. Progress in HDL-based therapies for atherosclerosis. Curr. Atheroscler. Rep. 3, 405–412 (2011)
    Article Google Scholar
  13. Alberti, K. G. et al. Harmonizing the metabolic syndrome: a joint interim statement of the International Diabetes Federation Task Force on Epidemiology and Prevention; National Heart, Lung, and Blood Institute; American Heart Association; World Heart Federation; International Atherosclerosis Society; and International Association for the Study of Obesity. Circulation 120, 1640–1645 (2009)
    Article CAS Google Scholar
  14. Wagner, J. E. et al. Old world nonhuman primate models of type 2 diabetes mellitus. ILAR J. 47, 259–271 (2006)
    Article CAS Google Scholar
  15. Fitzgerald, M. L. et al. ATP-binding cassette transporter A1 contains an NH2-terminal signal anchor sequence that translocates the protein’s first hydrophilic domain to the exoplasmic space. J. Biol. Chem. 276, 15137–15145 (2001)
    Article CAS Google Scholar
  16. Yvan-Charvet, L. et al. ATP-binding cassette transporters and HDL suppress hematopoietic stem cell proliferation. Science 328, 1689–1693 (2010)
    Article ADS CAS Google Scholar
  17. Kieft, K. A., Bocan, T. M. A. & Krause, B. R. Rapid on-line determination of cholesterol distribution among plasma lipoproteins after high-performance gel filtration chromatography. J. Lipid Res. 32, 859–866 (1991)
    CAS PubMed Google Scholar
  18. Garber, D. W., Kulkarni, K. R. & Anantharamaiah, G. M. A sensitive and convenient method for lipoprotein profile analysis of individual mouse plasma samples. J. Lipid Res. 41, 1020–1026 (2000)
    CAS PubMed Google Scholar
  19. Jeyarajah, E. J., Cromwell, W. C. & Otvos, J. D. Lipoprotein particle analysis by nuclear magnetic resonance spectroscopy. Clin. Lab. Med. 26, 847–870 (2006)
    Article Google Scholar
  20. Koritnik, D. L. & Rudel, L. L. Measurement of apolipoprotein A-I concentration in nonhuman primate serum by enzyme-linked immunosorbent assay (ELISA). J. Lipid Res. 24, 1639–1645 (1983)
    CAS PubMed Google Scholar

Download references

Acknowledgements

This work was supported by grants from the National Institutes of Health to K.J.M. (R01AG02055 and R01HL108182), E.A.F. (P01HL098055, R01HL084312 and R01HL58541), C.F.-H. (1P30HL101270 and R01HL107953), R.E.T. (R00HL088528), as well as by the Canadian Institutes of Health Research (K.J.R.)

Author information

Author notes

  1. Ryan E. Temel and Kathryn J. Moore: These authors contributed equally to this work.

Authors and Affiliations

  1. Leon H. Charney Division of Cardiology, Department of Medicine, Marc and Ruti Bell Vascular Biology and Disease Program, New York University School of Medicine, New York, 10016, New York, USA
    Katey J. Rayner, Farah N. Hussain, Janine M. van Gils, Tathagat D. Ray, Frederick J. Sheedy, Leigh Goedeke, Carlos Fernandez-Hernando, Edward A. Fisher & Kathryn J. Moore
  2. Regulus Therapeutics, San Diego, 92121, California, USA
    Christine C. Esau, Xueqing Liu, Oleg G. Khatsenko & Vivek Kaimal
  3. Department of Pathology-Section on Lipid Sciences, Wake Forest University School of Medicine, Winston-Salem, 27157, North Carolina, USA
    Allison L. McDaniel, Stephanie M. Marshall & Ryan E. Temel
  4. Department of Pathology-Section on Comparative Medicine, Wake Forest University School of Medicine, Winston-Salem, 27157, North Carolina, USA
    Cynthia J. Lees

Authors

  1. Katey J. Rayner
    You can also search for this author inPubMed Google Scholar
  2. Christine C. Esau
    You can also search for this author inPubMed Google Scholar
  3. Farah N. Hussain
    You can also search for this author inPubMed Google Scholar
  4. Allison L. McDaniel
    You can also search for this author inPubMed Google Scholar
  5. Stephanie M. Marshall
    You can also search for this author inPubMed Google Scholar
  6. Janine M. van Gils
    You can also search for this author inPubMed Google Scholar
  7. Tathagat D. Ray
    You can also search for this author inPubMed Google Scholar
  8. Frederick J. Sheedy
    You can also search for this author inPubMed Google Scholar
  9. Leigh Goedeke
    You can also search for this author inPubMed Google Scholar
  10. Xueqing Liu
    You can also search for this author inPubMed Google Scholar
  11. Oleg G. Khatsenko
    You can also search for this author inPubMed Google Scholar
  12. Vivek Kaimal
    You can also search for this author inPubMed Google Scholar
  13. Cynthia J. Lees
    You can also search for this author inPubMed Google Scholar
  14. Carlos Fernandez-Hernando
    You can also search for this author inPubMed Google Scholar
  15. Edward A. Fisher
    You can also search for this author inPubMed Google Scholar
  16. Ryan E. Temel
    You can also search for this author inPubMed Google Scholar
  17. Kathryn J. Moore
    You can also search for this author inPubMed Google Scholar

Contributions

K.J.M. and R.E.T. contributed equally to this study. K.J.M., R.E.T., C.C.E. and K.J.R. designed the study. C.J.L., R.E.T., A.L.M., S.M.M. and K.J.R. assisted in the necropsy. K.J.R., R.E.T., F.N.H., J.M.V.G., F.J.S., L.G. and T.D.R. performed the biological assays. C.C.E., X.L., O.G.K. and V.K. conducted the miRNA and microarray analyses. E.A.F. and C.F.-H. assisted with the experimental design and data interpretation. K.J.M. and K.J.R. wrote the first draft of the manuscript, which was commented on by all authors.

Corresponding authors

Correspondence toRyan E. Temel or Kathryn J. Moore.

Ethics declarations

Competing interests

E.A.F. is a Merck Advisory board member and receives honoraria for speaking engagements. C.C.E., X.L., O.G.K., V.K. are employees of Regulus Therapeutics. K.J.R., C.F-H. and K.J.M. have a pending patent on the use of miR-33 inhibitors.

Supplementary information

Supplementary Information

This file contains Supplementary Tables 1-3 and Supplementary Figures 1-4 with legends. (PDF 1306 kb)

PowerPoint slides

Rights and permissions

About this article

Cite this article

Rayner, K., Esau, C., Hussain, F. et al. Inhibition of miR-33a/b in non-human primates raises plasma HDL and lowers VLDL triglycerides.Nature 478, 404–407 (2011). https://doi.org/10.1038/nature10486

Download citation

This article is cited by

Editorial Summary

Manipulating plasma lipids

Recent work in mice has shown that microRNA-33a is an important regulator of lipid metabolism and that its inhibition can increase plasma high-density lipoprotein (HDL) and decrease atherosclerosis. Rayner et al. take an important step in translating these findings to non-human primates (African green monkeys), which, like humans and unlike mice, express both miR-33a and miR-33b. They find that anti-miR-33 is effective at inhibiting both miR-33a and miR-33b. As seen in the mouse studies, anti-miR33 raised plasma HDL but had the additional beneficial effect of reducing very low-density lipoprotein triglycerides, making this type of 'antagomir' therapy a candidate method of treating dyslipidaemias that increase cardiovascular disease risk.