Lentiviral shRNA knock-down of ADAMTS-5 and -9 restores matrix deposition in 3D chondrocyte culture.
Lentiviral shRNA knock-down of ADAMTS-5 and -9 restores matrix deposition in 3D chondrocyte culture.
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慢病毒 shRNA 敲低 ADAMTS-5 和 -9 可恢复 3D 软骨细胞培养物中的基质沉积。
DOI：
10.1002/term.275
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发表时间：
2010
影响因子：
3.3
通讯作者：
Coughlan TC
中科院分区：
文献类型：
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作者：
Coughlan TC
聚集蛋白聚糖是关节软骨的两种主要成分之一，并且在诸如骨关节炎（OA）的疾病期间，其受到蛋白水解酶的降解。负责聚集蛋白聚糖切割的主要蛋白酶是聚集蛋白聚糖酶，其被鉴定为蛋白酶ADAMTS家族的成员，其响应于炎症刺激而上调。目前还不确定六种聚集蛋白聚糖酶（ADAMTS-1、-4、-5、-8、-9和-15）中的哪一种主要负责人软骨中聚集蛋白聚糖的降解。在这里，我们发现六种聚集蛋白聚糖酶中的四种在永生化软骨细胞系中表达，并且可以响应于炎症细胞因子而上调。使用RNA干扰，我们证明了这些细胞中ADAMTS-5和-9表达的稳健敲低，并通过在三维（3D）支架上培养它们，表明ADAMTS-5酶表达的减少导致基质沉积的增加。这些数据表明，组织工程化软骨基质的质量可能会通过聚集蛋白聚糖酶表达的靶向耗竭来改善。此外，这项工作还提供了进一步的证据，表明ADAMTS-5可能是治疗关节炎疾病的治疗靶点。版权所有© 2010约翰威利父子有限公司.
Aggrecan is one of the two major constituents of articular cartilage, and during diseases such as osteoarthritis (OA) it is subject to degradation by proteolytic enzymes. The primary proteases responsible for aggrecan cleavage are the aggrecanases, identified as members of the ADAMTS family of proteases, which are upregulated in response to inflammatory stimuli. It is uncertain which of the six aggrecanases (ADAMTS‐1, ‐4, ‐5, ‐8, ‐9 and ‐15) are primarily responsible for the degradation of aggrecan in human cartilage. Here we show that four of the six aggrecanases are expressed in immortalized chondrocyte cell‐lines and can be upregulated in response to inflammatory cytokines. Using RNA interference, we demonstrate robust knock‐down of ADAMTS‐5 and ‐9 expression in these cells and, by culturing them on three‐dimensional (3D) scaffolds, show that reduction in expression of ADAMTS‐5 enzyme results in an increase in matrix deposition. These data suggest that the quality of tissue‐engineered cartilage matrix might be improved by targeted depletion of aggrecanase expression. Moreover, this work also provides further evidence that ADAMTS‐5 may be a therapeutic target in the treatment of arthritic disease. Copyright © 2010 John Wiley & Sons, Ltd.