Artificial MicroRNA-Mediated Tgfbr2 and Pdgfrb Co-Silencing Ameliorates Carbon Tetrachloride-Induced Hepatic Fibrosis in Mice.

Abstract

Hepatic stellate cells (HSCs) are the primary cell type responsible for liver fibrogenesis. Transforming growth factor beta 1 (TGF-β1) and platelet-derived growth factor (PDGF) are key profibrotic cytokines that regulate HSC activation and proliferation with functional convergence. Dual RNA interference against their receptors may achieve therapeutic effects. A novel RNAi strategy based on HSC-specific GFAP promoter-driven and lentiviral-expressed artificial microRNAs (amiRNAs) was devised that consists of an microRNA-30a backbone and effective shRNAs against mouse Pdgfrβ and Tgfbr2. Then, its antifibrotic efficacy was tested in primary and cultured HSCs and in mice affected with carbon tetrachloride-induced hepatic fibrosis. The study shows that amiRNA-mediated Pdgfrβ and Tgfbr2 co-silencing inhibits HSC activation and proliferation. After recombinant lentiviral particles were delivered into the liver via tail-vein injection, therapeutic amiRNAs were preferentially expressed in HSCs and efficiently co-knocked down in situ Tgfbr2 and Pdgfrβ expression, which correlates with downregulated expression of target or effector genes of their signaling, which include Pai-1, P70S6K, and D-cyclins. amiRNA-based HSC-specific co-silencing of Tgfbr2 and Pdgfrβ significantly suppressed hepatic expression of fibrotic markers α-Sma and Col1a1, extracellular matrix regulators Mmps and Timp1, and phenotypically ameliorated liver fibrosis, as indicated by reductions in serum alanine aminotransferase activity, collagen deposition, and α-Sma-positive staining. The findings provide proof of concept for the use of amiRNA-mediated co-silencing of two profibrogenic pathways in liver fibrosis treatment and highlight the therapeutic potential of concatenated amiRNAs for gene therapy.

• Authors: Yan Jiang, Yuanyuan Zhao, Fuchu He, Haijian Wang
• Journal: Human gene therapy (Hum Gene Ther) Vol. 30 Issue 2 Pg. 179-196 (02 2019) ISSN: 1557-7422 [Electronic] United States
• PMID: 30024280 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• MicroRNAs
• Receptor, Platelet-Derived Growth Factor beta
• Receptor, Transforming Growth Factor-beta Type II
• Tgfbr2 protein, mouse
• Tgfbr2 protein, rat

Topics

• Animals
• Carbon Tetrachloride Poisoning (genetics, metabolism, pathology, therapy)
• Cell Line
• Gene Silencing
• Hepatic Stellate Cells (metabolism, pathology)
• Liver Cirrhosis (genetics, metabolism, pathology, therapy)
• Mice
• MicroRNAs (biosynthesis, genetics)
• Rats
• Rats, Sprague-Dawley
• Receptor, Platelet-Derived Growth Factor beta (biosynthesis, genetics)
• Receptor, Transforming Growth Factor-beta Type II (biosynthesis, genetics)