Quantitative proteomics analysis of cartilage response to mechanical injury and cytokine treatment.
Quantitative proteomics analysis of cartilage response to mechanical injury and cytokine treatment.
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DOI：
10.1016/j.matbio.2016.12.004
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发表时间：
2017-11
期刊：
影响因子：
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通讯作者：
Önnerfjord P
中科院分区：
文献类型：
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作者：
Wang Y;Li Y;Khabut A;Chubinskaya S;Grodzinsky AJ;Önnerfjord P
据报道，关节损伤时的机械损伤和随后与滑液中促炎细胞因子水平升高相关的炎症反应有助于损伤后骨关节炎的进展。在这项探索性研究中，我们使用靶向蛋白质组学方法来跟踪基质降解对人膝关节软骨外植体的机械损伤和细胞因子治疗的反应，从而研究潜在的分子生物标志物。这种蛋白质组学方法使我们能够明确识别和量化软骨介质和外植体中的多种肽和蛋白质，这些肽和蛋白质在+/−损伤性压迫+/−细胞因子治疗后，模拟创伤后OA的最早期事件。我们跟踪了不同蛋白质结构域的降解，例如，G1/G2/G3的聚集蛋白聚糖，通过测量在整个21天培养期的7个时间点释放到培养基中的基质蛋白的代表性肽。先前在膝损伤/OA患者的滑液中鉴定的COMP neo表位也通过用cyt和cyt+inj处理的这些人软骨外植体释放。人类膝关节创伤后胶原蛋白前肽的缺乏和特异性COMP和COL 3A 1新表位水平的升高可能与创伤后OA的潜在生物标志物相关。因此，该模型系统能够研究软骨降解的动力学，并鉴定软骨外植体内的生物标志物和释放到培养基中的生物标志物。发现蛋白质组学揭示了在特定处理条件后鉴定的候选蛋白酶，包括MMP 1、MMP-3、MMP-10和MMP-13。
Mechanical damage at the time of joint injury and the ensuing inflammatory response associated with elevated levels of pro-inflammatory cytokines in the synovial fluid, are reported to contribute to the progression to osteoarthritis after injury. In this exploratory study, we used a targeted proteomics approach to follow the progression of matrix degradation in response to mechanical damage and cytokine treatment of human knee cartilage explants, and thereby to study potential molecular biomarkers. This proteomics approach allowed us to unambiguously identify and quantify multiple peptides and proteins in the cartilage medium and explants upon treatment with +/− injurious compression +/−cytokines, treatments that mimic the earliest events in post-traumatic OA. We followed degradation of different protein domains, e.g., G1/G2/G3 of aggrecan, by measuring representative peptides of matrix proteins released into the medium at 7 time points throughout the 21-day culture period. COMP neo epitopes, which were previously identified in the synovial fluid of knee injury/OA patients, were also released by these human cartilage explants treated with cyt and cyt+inj. The absence of collagen pro-peptides and elevated levels of specific COMP and COL3A1 neo-epitopes after human knee trauma may be relevant as potential biomarkers for post-traumatic OA. This model system thereby enables study of the kinetics of cartilage degradation and the identification of biomarkers within cartilage explants and those released to culture medium. Discovery proteomics revealed that candidate proteases were identified after specific treatment conditions, including MMP1, MMP-3, MMP-10 and MMP-13.