Guzzo Rosa M, Alaee Farhang, Paglia David, Gibson Jason D, Spicer Douglas, Drissi Hicham
Department of Orthopaedic Surgery, UConn Health, Farmington, CT, USA.
Stem Cell Institute, UConn Health, Farmington, CT, USA.
Genes Dis. 2016 Feb 24;3(1):88-99. doi: 10.1016/j.gendis.2015.12.005. eCollection 2016 Mar.
The bHLH transcription factor Twist1 has emerged as a negative regulator of chondrogenesis in skeletal progenitor cells and as an inhibitor of maturation in growth plate chondrocytes. However, its role in articular cartilage remains obscure. Here we examine Twist1 expression during re-differentiation of expanded human articular chondrocytes, the distribution of Twist1 proteins in normal versus OA human articular cartilage, and its role in modulating OA development in mice. High levels of Twist1 transcripts were detected by qPCR analyses of expanded de-differentiated human articular chondrocytes that had acquired mesenchymal-like features. The induction of hallmark cartilage genes by Bmp-2 mediated chondrogenic differentiation was paralleled by the dramatic suppression of Twist1 in vitro. In normal human articular cartilage, Twist1-expressing chondrocytes were most abundant in the superficial zone with little to no expression in the middle and deep zones. However, our analyses revealed a higher proportion of deep zone articular chondrocytes expressing Twist1 in human OA cartilage as compared to normal articular cartilage. Moreover, Twist1 expression was prominent within proliferative cell clusters near fissure sites in more severely affected OA samples. To assess the role of Twist1 in OA pathophysiology, we subjected wild type mice and transgenic mice with gain of Twist1 function in cartilage to surgical destabilization of the medial meniscus. At 12 weeks post-surgery, micro-CT and histological analyses revealed attenuation of the OA phenotype in Twist1 transgenic mice compared to wild type mice. Collectively, the data reveal a role for Twist in articular cartilage maintenance and the attenuation of cartilage degeneration.
bHLH转录因子Twist1已成为骨骼祖细胞软骨生成的负调节因子以及生长板软骨细胞成熟的抑制剂。然而,其在关节软骨中的作用仍不清楚。在此,我们研究了扩增的人关节软骨细胞重新分化过程中Twist1的表达、Twist1蛋白在正常与骨关节炎(OA)人关节软骨中的分布及其在调节小鼠OA发展中的作用。通过对已获得间充质样特征的扩增去分化人关节软骨细胞进行qPCR分析,检测到高水平的Twist1转录本。Bmp-2介导的软骨生成分化诱导标志性软骨基因的表达,与此同时,Twist1在体外被显著抑制。在正常人关节软骨中,表达Twist1的软骨细胞在表层区域最为丰富,而在中层和深层区域几乎不表达。然而,我们的分析显示,与正常关节软骨相比,在人OA软骨中,深层区域表达Twist1的关节软骨细胞比例更高。此外,在受影响更严重的OA样本中,裂隙部位附近的增殖细胞簇中Twist1表达显著。为了评估Twist1在OA病理生理学中的作用,我们对野生型小鼠和软骨中Twist1功能增强的转基因小鼠进行内侧半月板手术失稳处理。术后12周,显微CT和组织学分析显示,与野生型小鼠相比,Twist1转基因小鼠的OA表型有所减轻。总体而言,这些数据揭示了Twist在关节软骨维持和软骨退变减轻中的作用。