Fibroblasts from phenotypically normal palmar fascia exhibit molecular profiles highly similar to fibroblasts from active disease in Dupuytren's Contracture

Abstract

BackgroundDupuytren's contracture (DC) is a fibroproliferative disorder characterized by the progressive development of a scar-like collagen-rich cord that affects the palmar fascia of the hand and leads to digital flexion contractures. DC is most commonly treated by surgical resection of the diseased tissue, but has a high reported recurrence rate ranging from 27% to 80%. We sought to determine if the transcriptomic profiles of fibroblasts derived from DC-affected palmar fascia, adjacent phenotypically normal palmar fascia, and non-DC palmar fascial tissues might provide mechanistic clues to understanding the puzzle of disease predisposition and recurrence in DC.MethodsTo achieve this, total RNA was obtained from fibroblasts derived from primary DC-affected palmar fascia, patient-matched unaffected palmar fascia, and palmar fascia from non-DC patients undergoing carpal tunnel release (6 patients in each group). These cells were grown on a type-1 collagen substrate (to better mimic their in vivo environments). Microarray analyses were subsequently performed using Illumina BeadChip arrays to compare the transcriptomic profiles of these three cell populations. Data were analyzed using Significance Analysis of Microarrays (SAM v3.02), hierarchical clustering, concordance mapping and Venn diagram.ResultsWe found that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected fascia of DC patients exhibited a much greater overlap than fibroblasts derived from the palmar fascia of patients undergoing carpal tunnel release. Quantitative real time RT-PCR confirmed the differential expression of select genes validating the microarray data analyses. These data are consistent with the hypothesis that predisposition and recurrence in DC may stem, at least in part, from intrinsic similarities in the basal gene expression of diseased and phenotypically unaffected palmar fascia fibroblasts. These data also demonstrate that a collagen-rich environment differentially alters gene expression in these cells. In addition, Ingenuity pathway analysis of the specific biological pathways that differentiate DC-derived cells from carpal tunnel-derived cells has identified the potential involvement of microRNAs in this fibroproliferative disorder.ConclusionsThese data show that the transcriptomic profiles of DC-disease fibroblasts and fibroblasts from unaffected palmar fascia in DC patients are highly similar, and differ significantly from the transcriptomic profiles of fibroblasts from the palmar fascia of patients undergoing carpal tunnel release.

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

“…The cellular and molecular physiology underlying this trend to recurrence remains under investigation. Our work has shown that not only fibroblasts from actively diseased cords, but also fibroblasts from phenotypically normal adjacent palmar fascia in DD have significantly altered patterns of gene expression compared to control cells from CT patients [37]. These nearby cells may therefore represent a residual manifestation of proto-disease after surgery, which may be stimulated by cytokine mediators to progress to active, frank recurrence.…”

Unraveling the signaling pathways promoting fibrosis in Dupuytren's disease reveals TNF as a therapeutic target

“…Conversely, LiCl led to increased TOPFlash activity in nonpalmar fibroblasts from Dupuytren's patients and palmar dermal fibroblasts from individuals unaffected with Dupuytren's disease. It has previously been shown that fibroblasts from different sources may show divergent responses to the same stimuli (43), and more recently (29), it was reported that Dupuytren's myofibroblasts and fibroblasts from uninvolved palmar fascia from the same patients both expressed genes related to Wnt/β-catenin pathway, unlike fascial-derived fibroblasts from individuals without Dupuytren's disease.…”

“…However, this approach has limitations. The palmar fascia over the carpal tunnel is rarely affected by Dupuytren's disease in susceptible individuals, and the transverse carpal ligament is always unaffected; hence, it is possible that the constituent cells are inherently different (29). Furthermore, with the exception of nodules in Dupuytren's disease, fascia is sparsely populated by cells and hence to obtain adequate numbers, most authors use cells to passage 5 (12).…”

Down‐regulation of collagen synthesis and matrix metalloproteinase expression in myofibroblasts from dupuytren nodule using adenovirus‐mediated relaxin gene therapy

“…The expression of collagenases and their inhibitors in the normal ligament differs from that seen in the palmar fascia and nodule ligaments from Dupuytren's disease. The synthesis of MMPs and TIMP 1 is increased, despite the down‐regulation of TIMP3 and 4 in the mRNA profile in the myofibroblasts from Dupuytren's patients, as compared to normal ligament …”