Targeting CCN2 protects against progressive non-alcoholic steatohepatitis in a preclinical model induced by high-fat feeding and type 2 diabetes

Jing Ren; Xiaoyu Wang; Sarah N Parry; Christine Yee; Mark D Gorrell; Susan V McLennan; Stephen M Twigg

doi:10.1007/s12079-022-00667-1

Targeting CCN2 protects against progressive non-alcoholic steatohepatitis in a preclinical model induced by high-fat feeding and type 2 diabetes

J Cell Commun Signal. 2022 Sep;16(3):447-460. doi: 10.1007/s12079-022-00667-1. Epub 2022 Jan 17.

Authors

Jing Ren¹, Xiaoyu Wang¹, Sarah N Parry^{1

2}, Christine Yee¹, Mark D Gorrell^{3

4}, Susan V McLennan^{1

2

5}, Stephen M Twigg^{6

7}

Affiliations

¹ Greg Brown Diabetes and Endocrinology Research Laboratories, Sydney Medical School (Central), Faculty of Medicine and Health, Charles Perkins Centre, The University of Sydney, Sydney, NSW, 2006, Australia.
² Department of Endocrinology, Royal Prince Alfred Hospital, Camperdown, NSW, 2050, Australia.
³ Centenary Institute, The University of Sydney, Camperdown, NSW, 2050, Australia.
⁴ Faculty of Medicine and Health, The University of Sydney, Sydney, NSW, 2006, Australia.
⁵ New South Wales Health Pathology, Camperdown, NSW, 2050, Australia.
⁶ Greg Brown Diabetes and Endocrinology Research Laboratories, Sydney Medical School (Central), Faculty of Medicine and Health, Charles Perkins Centre, The University of Sydney, Sydney, NSW, 2006, Australia. stephen.twigg@sydney.edu.au.
⁷ Department of Endocrinology, Royal Prince Alfred Hospital, Camperdown, NSW, 2050, Australia. stephen.twigg@sydney.edu.au.

Abstract

Type 2 diabetes is an independent risk factor for non-alcoholic steatohepatitis (NASH) progression and its mediators have not been resolved. In this study, a pathogenic role of cellular communication network factor 2 (CCN2) protein in NASH pathology, was investigated in an established preclinical NASH model. Male wild type C57BL/6 mice received either Chow or high fat diet (HFD) for 26 weeks, with some mice in each group randomly selected to receive low dose streptozotocin (STZ: 3 i.p. injections, 65 mg/kg) at 15 weeks to induce type 2 diabetes. In the final 10 of the 26 weeks mice from each group were administered i.p. either rabbit anti-CCN2 neutralizing antibody (CCN2Ab) or as control normal rabbit IgG, at a dose of 150 µg per mouse twice/week. NASH developed in the HFD plus diabetes (HFD+DM) group. Administration of CCN2Ab significantly downregulated collagen I and collagen III mRNA induction and prevented pro-inflammatory MCP-1 mRNA induction in HFD+DM mice. At the protein level, CCN2Ab significantly attenuated collagen accumulation by PSR stain and collagen I protein induction in HFD+DM. Phosphorylation of the pro-fibrotic ERK signalling pathway in liver in HFD+DM was attenuated by CCN2Ab treatment. Intrahepatic CCN1 mRNA was induced, whereas CCN3 was downregulated at both the mRNA and protein levels in HFD+DM. CCN3 down-regulation was prevented by CCN2Ab treatment. This in vivo study indicates that CCN2 is a molecular target in NASH with high fat diet and diabetes, and that regulation of ERK signalling is implicated in this process.

Keywords: CCN2; Fibrosis; Inflammation; NAFLD; Neutralizing antibody; Type 2 diabetes.