Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, William Henry Duncan Building, 6 West Derby Street, Liverpool L7 8TX, United Kingdom.
Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, William Henry Duncan Building, 6 West Derby Street, Liverpool L7 8TX, United Kingdom.
J Biol Chem. 2019 Aug 30;294(35):13027-13039. doi: 10.1074/jbc.RA118.006865. Epub 2019 Jul 12.
Osteoarthritis is a chronic disease characterized by the loss of articular cartilage in synovial joints through a process of extracellular matrix destruction that is strongly associated with inflammatory stimuli. Chondrocytes undergo changes to their protein translational capacity during osteoarthritis, but a study of how disease-relevant signals affect chondrocyte protein translation at the transcriptomic level has not previously been performed. In this study, we describe how the inflammatory cytokine interleukin 1-β (IL-1β) rapidly affects protein translation in the chondrocytic cell line SW1353. Using ribosome profiling we demonstrate that IL-1β induced altered translation of inflammatory-associated transcripts such as NFKB1, TNFAIP2, MMP13, CCL2, and CCL7, as well as a number of ribosome-associated transcripts, through differential translation and the use of multiple open reading frames. Proteomic analysis of the cellular layer and the conditioned media of these cells identified changes in a number of the proteins that were differentially translated. Translationally regulated secreted proteins included a number of chemokines and cytokines, underlining the rapid, translationally mediated inflammatory cascade that is initiated by IL-1β. Although fewer cellular proteins were found to be regulated in both ribosome profiling and proteomic data sets, we did find increased levels of SOD2, indicative of redox changes within SW1353 cells being modulated at the translational level. In conclusion, we have produced combined ribosome profiling and proteomic data sets that provide a valuable resource in understanding the processes that occur during cytokine stimulation of chondrocytic cells.
骨关节炎是一种慢性疾病，其特征是滑膜关节中的关节软骨通过细胞外基质破坏过程而丧失，这一过程与炎症刺激密切相关。在骨关节炎期间，软骨细胞的蛋白质翻译能力会发生变化，但此前尚未在转录组水平上研究与疾病相关的信号如何影响软骨细胞的蛋白质翻译。在本研究中，我们描述了炎性细胞因子白细胞介素1-β（IL-1β）如何快速影响软骨细胞系SW1353中的蛋白质翻译。使用核糖体分析，我们证明IL-1β通过差异翻译和使用多个开放阅读框，诱导了炎性相关转录本如NFKB1、TNFAIP2、MMP13、CCL2和CCL7以及一些核糖体相关转录本的翻译改变。对这些细胞的细胞层和条件培养基进行蛋白质组分析，确定了许多差异翻译的蛋白质的变化。翻译调控的分泌蛋白包括多种趋化因子和细胞因子，突显了由IL-1β引发的快速的、翻译介导的炎症级联反应。尽管在核糖体分析和蛋白质组数据集中发现受调控的细胞蛋白较少，但我们确实发现超氧化物歧化酶2（SOD2）水平升高，这表明SW1353细胞内的氧化还原变化在翻译水平上受到调节。总之，我们生成了核糖体分析和蛋白质组数据集，为理解细胞因子刺激软骨细胞过程中发生的机制提供了宝贵资源。
