Knockdown of vimentin reduces mesenchymal phenotype of cholangiocytes in the Mdr2−/− mouse model of primary sclerosing cholangitis (PSC)

Abstract

BackgroundCholangiocytes are the target cells of cholangiopathies including primary sclerosing cholangitis (PSC). Vimentin is an intermediate filament protein that has been found in various types of mesenchymal cells. The aim of this study is to evaluate the role of vimentin in the progression of biliary damage/liver fibrosis and whether there is a mesenchymal phenotype of cholangiocytes in the Mdr2−/− model of PSC.MethodsIn vivo studies were performed in 12 wk. Mdr2−/− male mice with or without vimentin Vivo-Morpholino treatment and their corresponding control groups. Liver specimens from human PSC patients, human intrahepatic biliary epithelial cells (HIBEpiC) and human hepatic stellate cell lines (HHSteCs) were used to measure changes in epithelial-to-mesenchymal transition (EMT).FindingsThere was increased mesenchymal phenotype of cholangiocytes in Mdr2−/− mice, which was reduced by treatment of vimentin Vivo-Morpholino. Concomitant with reduced vimentin expression, there was decreased liver damage, ductular reaction, biliary senescence, liver fibrosis and TGF-β1 secretion in Mdr2−/− mice treated with vimentin Vivo-Morpholino. Human PSC patients and derived cell lines had increased expression of vimentin and other mesenchymal markers compared to healthy controls and HIBEpiC, respectively. In vitro silencing of vimentin in HIBEpiC suppressed TGF-β1-induced EMT and fibrotic reaction. HHSteCs had decreased fibrotic reaction and increased cellular senescence after stimulation with cholangiocyte supernatant with reduced vimentin levels.InterpretationOur study demonstrated that knockdown of vimentin reduces mesenchymal phenotype of cholangiocytes, which leads to decreased biliary senescence and liver fibrosis. Inhibition of vimentin may be a key therapeutic target in the treatment of cholangiopathies including PSC.FundNational Institutes of Health (NIH) awards, VA Merit awards.

Combination of single cell sequencing data and GWAS summary statistics reveals genetically-influenced liver cell types for primary biliary cholangitis

“…We found that cholangiocytes showed a notable enrichment by PBC-relevant genetic association signals (Permuted P < 0.05, Figure 3b and Supplemental Table S10). These results are consistent with previous evidence that an immune-mediated injury of cholangiocytes contributes risk to PBC 44,45 .…”

Impact of Aging on Liver Cells and Liver Disease: Focus on the Biliary and Vascular Compartments

“…( 42 ) Other correlative assessments regarding α‐CGRP, SCF, or vimentin levels were carried out in PSC human tissue to confirm the preclinical data obtained in rodents, as previously described. ( 49 , 50 , 51 ) Finally, increased levels of twinfilin‐1 were detected in cholangiocytes of patients with PBC or PSC. This protein, in experimental cholestasis, appeared to respond to several microRNA signals and to play a major role in supporting the proliferative/senescent evolution of bile duct cells in this setting.…”

“…Its inhibition improves biliary damage and cellular senescence, possibly interfering with the epithelial to mesenchymal transition (EMT) process of bile duct cells, a mechanism linked to the onset of fibrosis. ( 51 ) Finally, the direct targeting of senescent cellular machinery has been attempted, in experimental systems, to improve pathological features. Inhibition of the cyclin‐dependent kinase inhibitor p16 has been obtained in a model of cholestasis by the administration of p16 morpholino.…”

Pan-Cancer Analysis of VIM Expression in Human Cancer Tissues

“…A growing body of literature indicates that vimentin controls cell proliferation (Cheng 2016). Extensive research in vimentin-de cient animal models (for example, vim −/− mice) showed that loss of vimentin caused a reduction in brosis and the mesenchymal phenotype of cells (for example, in cholangiocytes), which could be reversed upon vim re-expression (Eckes 1998;Zhou 2019). In contrast, some oncogenes could increase cell proliferation, as a result of a higher vimentin messenger RNA (mRNA) and protein levels (Rathje 2014).…”