Identification of differentially expressed genes in fibroblasts derived from patients with Dupuytren's Contracture

Dupuytren's contracture (DC) is the most common inherited connective tissue disease of humans and is hypothesized to be associated with aberrant wound healing of the palmar fascia. Fibroblasts and myofibroblasts are believed to play an important role in the genesis of DC and the fibroproliferation and contraction that are hallmarks of this disease. This study compares the gene expression profiles of fibroblasts isolated from DC patients and controls in an attempt to identify key genes whose regulation might be significantly altered in fibroblasts found within the palmar fascia of Dupuytren's patients. Total RNA isolated from diseased palmar fascia (DC) and normal palmar fascia (obtained during carpal tunnel release; 6 samples per group) was subjected to quantitative analyses using two different microarray platforms (GE Code Link™ and Illumina™) to identify and validate differentially expressed genes. The data obtained was analyzed using The Significance Analysis of Microarrays (SAM) software through which we identified 69 and 40 differentially regulated gene transcripts using the CodeLink™ and Illumina™ platforms, respectively. The CodeLink™ platform identified 18 upregulated and 51 downregulated genes. Using the Illumina™ platform, 40 genes were identified as downregulated, eleven of which were identified by both platforms. Quantitative RT-PCR confirmed the downregulation of three high-interest candidate genes which are all components of the extracellular matrix: proteoglycan 4 (PRG4), fibulin-1 (FBLN-1) transcript variant D, and type XV collagen alpha 1 chain. Overall, our study has identified a variety of candidate genes that may be involved in the pathophysiology of Dupuytren's contracture and may ultimately serve as attractive molecular targets for alternative therapies. Dupuytren's contracture and may ultimately serve as attractive molecular targets for alternative therapies. 6 Samples from carpal-tunnel derived fibroblasts and 6 Samples from Dupuytren's contracture-derived fibroblasts were hybridized to Illumina's Sentrix Human-6 Expression Beadchip and GE Code Link arrays.

杜普伊特伦挛缩症（Dupuytren's contracture, DC）是人类最常见的遗传性结缔组织疾病，现有假说认为其与掌腱膜的异常伤口愈合相关。成纤维细胞（fibroblasts）与肌成纤维细胞（myofibroblasts）被认为在DC的发病机制，以及该病标志性的纤维增生与挛缩过程中发挥重要作用。本研究对源自DC患者与健康对照者的成纤维细胞基因表达谱进行比较，旨在鉴定出杜普伊特伦挛缩症患者掌腱膜内成纤维细胞中调控模式发生显著改变的关键基因。研究人员从病变掌腱膜（DC组）与正常掌腱膜（取自腕管松解术，每组6例样本）中提取总RNA，采用两种不同的微阵列平台：GE Code Link™与Illumina™，进行定量分析以鉴定并验证差异表达基因。所得数据通过微阵列显著性分析（Significance Analysis of Microarrays, SAM）软件进行分析，分别借助CodeLink™与Illumina™平台鉴定出69条与40条差异调控的基因转录本。其中CodeLink™平台鉴定出18条上调基因与51条下调基因；Illumina™平台则鉴定出40条下调基因，其中11条可被两个平台共同识别。定量逆转录聚合酶链反应验证了3个高关注度候选基因的下调表达，这3个基因均为细胞外基质成分：蛋白聚糖4（PRG4）、纤连蛋白-1（FBLN-1）转录变体D，以及XV型胶原α1链。总体而言，本研究鉴定出多种可能参与杜普伊特伦挛缩症病理生理过程的候选基因，这些基因有望成为替代疗法的潜在分子靶点。杜普伊特伦挛缩症并有望成为替代疗法的潜在分子靶点。本研究共纳入6例腕管来源成纤维细胞样本与6例杜普伊特伦挛缩症来源成纤维细胞样本，将其分别杂交至Illumina Sentrix Human-6表达微珠芯片与GE Code Link微阵列。