Antisense oligonucleotides targeting hepatic angiotensinogen reduce atherosclerosis and liver steatosis in hypercholesterolemic mice

Dien Ye^1,2, Congqing Wu^1,3,4,5, Lei Cai¹, Deborah A. Howatt¹, Ching-Ling Liang¹, Yuriko Katsumata^6,7, Adam E. Mullick⁸, Ryan E. Temel^1,9, A.H. Jan Danser², Alan Daugherty^1,3,9, Hong S. Lu^1,3,9*

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¹ Saha Cardiovascular Research Center, University of Kentucky, Lexington, KY, USA

² Division of Vascular Medicine and Pharmacology, Department of Internal Medicine, Erasmus MC, Rotterdam, Netherlands

³ Saha Aortic Center, University of Kentucky, Lexington, KY, USA

⁴ Department of Surgery, University of Kentucky, Lexington, KY, USA

⁵ Department of Microbiology, Immunology, and Molecular Genetics, University of Kentucky, Lexington, KY, USA

⁶ Department of Biostatistics, College of Public Health, University of Kentucky, Lexington, KY, USA

⁷ Sanders-Brown Center on Aging, University of Kentucky, Lexington, KY, USA

⁸ Ionis Pharmaceuticals, Inc., Carlsbad, CA, USA

⁹ Department of Physiology, University of Kentucky, Lexington, KY, USA

Global Translational Medicine 2023, 2(1), 288 https://doi.org/10.36922/gtm.288

Submitted: 8 December 2022 | Accepted: 7 February 2023 | Published: 24 February 2023

© 2023 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )

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Abstract

Hepatocyte-derived angiotensinogen (AGT) is the precursor of angiotensin II (AngII). We determined the effects of hepatocyte-specific (N-acetylgalactosamine-conjugated) antisense oligonucleotides targeting AGT (GalNAc AGT ASO) on AngII-mediated blood pressure (BP) regulation and atherosclerosis and compared its effects with losartan, an AngII type 1 (AT1) receptor blocker, in hypercholesterolemic mice. Eight-week-old male low-density lipoprotein (LDL) receptor deficient mice were administered vehicle or GalNAc AGT ASO (1, 2.5, or 5 mg/kg) subcutaneously beginning 2 weeks before the initiation of Western diet feeding. All mice were fed Western diet for 12 weeks. Their systolic BP was monitored by the tail-cuff technique, and the atherosclerotic lesion area was measured by an en face method. Although the effects of all 3 doses of GalNAc AGT ASO on plasma AGT concentrations were similar, GalNAc AGT ASO reduced BP and atherosclerotic lesion size in a dose-dependent manner. Subsequently, we compared the effects of GalNAc AGT ASO (5 mg/kg) with losartan (15 mg/kg/day). Compared to losartan, GalNAc AGT ASO led to more profound increases in plasma renin and reduction in BP but had similar effects on atherosclerosis. Remarkably, GalNAc AGT ASO also reduced liver steatosis, which was not observed in losartan-treated mice. In conclusion, the BP increase and atherosclerosis development in hypercholesterolemic mice are dependent on AngII generated from hepatic AGT. Deleting hepatic AGT improves diet-induced liver steatosis, and this occurs in an AT1 receptor-independent manner.

Keywords

Angiotensinogen

Antisense oligonucleotides

Blood pressure

Liver steatosis

Atherosclerosis

Funding

National Heart, Lung, and Blood Institute of the National Institutes of Health

Conflict of interest

Alan Daugherty and Hong S. Lu have submitted a patent application for use of antisense oligonucleotides targeting angiotensinogen in thoracic aortic aneurysms. Adam E. Mullick is an employee of Ionis Pharmaceuticals, who provided the mouse GalNAc AGT ASO. IONIS-AGT-Lrx and ION904 are both AGT ASO Ionis programs in clinical development. Ionis Pharmaceuticals did not provide any funding for this study. The other authors report no conflicts.

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