Wilkinson Janine M, Davidson Rose K, Swingler Tracey E, Jones Eleanor R, Corps Anthony N, Johnston Phillip, Riley Graham P, Chojnowski Adrian J, Clark Ian M
School of Biological Sciences, University of East Anglia, Norwich Research Park, NR4 7TJ, UK.
Biochim Biophys Acta. 2012 Jun;1822(6):897-905. doi: 10.1016/j.bbadis.2012.02.001. Epub 2012 Feb 9.
Dupuytren's disease (DD) is a common fibrotic condition of the palmar fascia, leading to deposition of collagen-rich cords and progressive flexion of the fingers. The molecular mechanisms underlying the disease are poorly understood. We have previously shown altered expression of extracellular matrix-degrading proteases (matrix metalloproteases, MMPs, and 'a disintegrin and metalloprotease domain with thrombospondin motifs', ADAMTS, proteases) in palmar fascia from DD patients compared to control and shown that the expression of a sub-set of these genes correlates with post-operative outcome. In the current study we used an in vitro model of collagen contraction to identify the specific proteases which mediate this effect. We measured the expression of all MMPs, ADAMTSs and their inhibitors in fibroblasts derived from the palmar fascia of DD patients, both in monolayer culture and in the fibroblast-populated collagen lattice (FPCL) model of cell-mediated contraction. Key proteases, previously identified in our tissue studies, were expressed in vitro and regulated by tension in the FPCL, including MMP1, 2, 3, 13 and 14. Knockdown of MMP2 and MMP14 (but not MMP1, 3 and 13) inhibited cell-mediated contraction, and knockdown of MMP14 inhibited proMMP-2 activation. Interestingly, whilst collagen is degraded during the FPCL assay, this is not altered upon knockdown of any of the proteases examined. We conclude that MMP-14 (via its ability to activate proMMP-2) and MMP-2 are key proteases in collagen contraction mediated by fibroblasts in DD patients. These proteases may be drug targets or act as biomarkers for disease progression.
掌腱膜挛缩症（DD）是一种常见的掌腱膜纤维化疾病，会导致富含胶原蛋白的条索状沉积以及手指的渐进性屈曲。该疾病的分子机制尚不清楚。我们之前已经表明，与对照组相比，DD患者掌腱膜中细胞外基质降解蛋白酶（基质金属蛋白酶、MMPs，以及具有血小板反应蛋白基序的解整合素和金属蛋白酶结构域，ADAMTSs，蛋白酶）的表达发生了改变，并且这些基因的一个子集的表达与术后结果相关。在当前研究中，我们使用了胶原收缩的体外模型来鉴定介导这种效应的特定蛋白酶。我们测量了DD患者掌腱膜来源的成纤维细胞中所有MMPs、ADAMTSs及其抑制剂的表达，这些细胞处于单层培养以及细胞介导收缩的成纤维细胞填充胶原晶格（FPCL）模型中。我们在组织研究中之前鉴定出的关键蛋白酶在体外表达，并在FPCL中受张力调节，包括MMP1、2、3、13和14。敲低MMP2和MMP14（但不是MMP1、3和13）会抑制细胞介导的收缩，并且敲低MMP14会抑制proMMP - 2的激活。有趣的是，虽然在FPCL测定过程中胶原蛋白会被降解，但在敲低任何所检测的蛋白酶后，这种降解并未改变。我们得出结论，MMP - 14（通过其激活proMMP - 2的能力）和MMP - 2是DD患者成纤维细胞介导的胶原收缩中的关键蛋白酶。这些蛋白酶可能是药物靶点或作为疾病进展的生物标志物。
