Increased CCT-eta expression is a marker of latent and active disease and a modulator of fibroblast contractility in Dupuytren’s contracture

Abstract

Dupuytren's contracture (DC) is a fibroproliferative disorder of unknown etiology characterized by a scar-like contracture that develops in the palm and/or digits. We have previously reported that the eta subunit of the chaperonin containing T-complex polypeptide (CCT-eta) is increased in fibrotic wound healing, and is essential for the accumulation of α-smooth muscle actin (α-SMA) in fibroblasts. The purpose of this study was to determine if CCT-eta is similarly implicated in the aberrant fibrosis seen in DC and to investigate the role of CCT-eta in the behavior of myo/fibroblasts in DC. Fibroblasts were obtained from DC-affected palmar fascia, from adjacent phenotypically normal palmar fascia in the same DC patients (PF), and from non-DC palmar fascial tissues in patients undergoing carpal tunnel (CT) release. Inherent contractility in these three populations was examined using fibroblast-populated collagen lattices (FPCLs) and by cell traction force microscopy. Expression of CCT-eta and α-SMA protein was determined by Western blot. The effect of CCT-eta inhibition on the contractility of DC cells was determined by deploying an siRNA versus CCT-eta. DC cells were significantly more contractile than both matching palmar fascial (PF) cells and CT cells in both assays, with PF cells demonstrating an intermediate contractility in the FPCL assay. Whereas α-SMA protein was significantly increased only in DC cells compared to PF and CT cells, CCT-eta protein was significantly increased in both PF and DC cells compared to CT cells. siRNA-mediated depletion of CCT-eta inhibited the accumulation of both CCT-eta and α-SMA protein in DC cells, and also significantly decreased the contractility of treated DC cells. These observations suggest that increased expression of CCT-eta appears to be a marker for latent and active disease in these patients and to be essential for the increased contractility exhibited by these fibroblasts.

Anti-fibrotic action of pirfenidone in Dupuytren’s disease-derived fibroblasts

“…Cell contractility of the fibroblasts was tested using FPCL assay as previously described with slight modifications [32]. In brief, collagen lattices were prepared by mixing cell suspensions with a neutralized solution of type I collagen matrix (four parts rat tail collagen, one part neutralization solution; two parts 0.34 M NaOH: 3 parts 10X Waymouth media).…”

Comprehensive analysis of the expression of chaperonin containing TCP1 subunits (CCTs) and their influence on prognosis in hepatocellular carcinoma

“…siRNA-mediated depletion of CCT7 inhibited the accumulation of α-SMA protein in Dupuytren’s contracture (DC) cells and significantly decreased the contractility of treated DC cells ( 29 ). In this study, the expression of CCT7 was upregulated in HCC tissues compared with the expression in normal liver tissues and correlated with AFP, pathological grade, and macro- and microvascular invasion but not with the Child-Pugh classification.…”

Integration of Airway Inflammation and Remodeling Mechanisms Specific to Eosinophilic Asthma Through Differential Co-Expression of Genes in Bronchial Brush Biopsy Samples

“…Its expression increases in brotic wound healing and is essential for the accumulation of α-smooth muscle actin (α-SMA) in broblasts and cell differentiation to myo broblasts 80 . CCT7 was recently found to coimmunoprecipitate with thromboxane A2 receptor as well as β2-adrenergic receptor from human HEK 293 cells and its depletion resulted in reduced cell surface expression of both receptors 81 ; a fact signi cant given the momentous role of both G-protein coupled receptors in asthma pathogenesis.…”