1. Identification of Novel Fibrosis Modifiers by In Vivo siRNA Silencing
- Author
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Vollmann, Elisabeth H, Cao, Lizhi, Amatucci, Aldo, Reynolds, Taylor, Hamann, Stefan, Dalkilic-Liddle, Isin, Cameron, Thomas O, Hossbach, Markus, Kauffman, Kevin J, Mir, Faryal F, Anderson, Daniel G, Novobrantseva, Tatiana, Koteliansky, Victor, Kisseleva, Tatiana, Brenner, David, Duffield, Jeremy, and Burkly, Linda C
- Subjects
LNP ,Liver Disease ,fibrosis ,Chronic Liver Disease and Cirrhosis ,Clinical Sciences ,myofibroblast ,siRNA ,in vivo gene silencing ,Genetics ,2.1 Biological and endogenous factors ,Egr2 ,Biochemistry and Cell Biology ,Aetiology ,Digestive Diseases ,Biotechnology - Abstract
Fibrotic diseases contribute to 45% of deaths in the industrialized world, and therefore a better understanding of the pathophysiological mechanisms underlying tissue fibrosis is sorely needed. We aimed to identify novel modifiers of tissue fibrosis expressed by myofibroblasts and their progenitors in their disease microenvironment through RNA silencing invivo. We leveraged novel biology, targeting genes upregulated during liver and kidney fibrosis in this cell lineage, and employed small interfering RNA (siRNA)-formulated lipid nanoparticles technology to silence these genes in carbon-tetrachloride-induced liver fibrosis in mice. We identified five genes, Egr2, Atp1a2, Fkbp10, Fstl1, and Has2, which modified fibrogenesis based on their silencing, resulting in reduced Col1a1 mRNA levels and collagen accumulation in the liver. These genes fell intodifferent groups based on the effects of their silencing on a transcriptional mini-array and histological outcomes. Silencing of Egr2 had the broadest effects invivo and also reduced fibrogenic gene expression in a human fibroblast cell line. Prior to our study, Egr2, Atp1a2, and Fkbp10 had not been functionally validated in fibrosis invivo. Thus, our results provide a major advance over the existing knowledge of fibrogenic pathways. Our study is the first example of a targeted siRNA assay to identify novel fibrosis modifiers invivo.
- Published
- 2017