Bromodomain-containing-protein-4 and cyclin-dependent-kinase-9 inhibitors interact synergistically in vitro and combined treatment reduces post-traumatic osteoarthritis severity in mice.
Bromodomain-containing-protein-4 and cyclin-dependent-kinase-9 inhibitors interact synergistically in vitro and combined treatment reduces post-traumatic osteoarthritis severity in mice.
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DOI：
10.1016/j.joca.2020.07.012
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发表时间：
2021-01
影响因子：
7
通讯作者：
Haudenschild DR
中科院分区：
文献类型：
--
作者：
Fukui T;Yik JHN;Doyran B;Davis J;Haudenschild AK;Adamopoulos IE;Han L;Haudenschild DR
关节损伤迅速诱导初级反应基因(PRGS)的表达，这些初级反应基因激活一系列次级基因，破坏关节组织，引发创伤后骨关节炎(PTOA)。溴域蛋白-4(Brd4)和细胞周期蛋白依赖蛋白-9(CDK9)共同控制PRG反式激活的限速步骤，包括促炎基因。这项研究调查了Brd4和CDK9抑制剂在体外和小鼠PTOA模型中是否抑制炎症和防止PTOA的发展。在不同的环境下，严格测试Brd4和CDK9抑制剂(JQ1和Framopiridol)对PRG及其相关的二次损伤的影响。用RT-β和基因芯片检测炎症刺激对人软骨细胞PRG表达的短期影响。我们通过RT-β(2-24小时)、体内基质金属蛋白酶活性成像(24小时)、原子力显微镜纳米压痕(3-7天)和组织学(3天-4周)对IL-1处理的牛软骨移植块3-6天后释放的糖胺多糖和小鼠前交叉韧带断裂后骨关节的变化进行了量化。通过基因芯片分析，黄吡哆醇和JQ1抑制剂具有协同作用，两者的组合几乎完全阻止了体外大多数IL-1β诱导的PRG的激活，并阻止了IL-1β诱导的糖胺多糖从软骨外植体中释放。。与未治疗的前交叉韧带断裂小鼠相比，联合用药组小鼠的IL-1β和IL-6表达减少，体内基质金属蛋白酶活性降低，滑膜炎(1.5vs.4.9)和OARSI评分(2.8vs.6.0)降低。JQ1和黄吡哆醇在减少关节创伤后的损伤反应方面具有协同作用，提示靶向Brd4和/或CDK9可能是PTOA预防和治疗早期OA的可行策略。
Joint injury rapidly induces expression of primary response genes (PRGs), which activate a cascade of secondary genes that destroy joint tissues and initiate post-traumatic osteoarthritis (PTOA). Bromodomain-containing-protein-4 (Brd4) and cyclin-dependent-kinase-9 (CDK9) cooperatively control the rate-limiting step of PRG transactivation, including pro-inflammatory genes. This study investigated whether Brd4 and CDK9 inhibitors suppress inflammation and prevent PTOA development in-vitro and in a mouse PTOA model. The effects of Brd4 and CDK9 inhibitors (JQ1 and Flavopiridol) on PRG and associated secondary damage were rigorously tested in different settings. Short-term effects of inflammatory stimuli (IL-1β, IL-6, TNF) on human chondrocyte PRG expression were assessed by RT-PCR and microarray after 5-hours. We quantified glycosaminoglycan release from IL-1β-treated bovine cartilage explants after 3-6 days, and osteoarthritic changes in mice after ACL-rupture using RT-PCR (2-24hrs), in-vivo imaging of MMP activity (24hrs), AFM-nanoindentation (3-7days), and histology (3days-4wks). Flavopiridol and JQ1 inhibitors act synergistically, and a combination of both almost completely prevented the activation of most IL-1β-induced PRGs in-vitro by microarray analysis, and prevented IL-1β-induced glycosaminoglycan release from cartilage explants. . Mice given the drug combination showed reduced IL-1β and IL-6 expression, less in-vivo MMP activity, and lower synovitis (1.5 vs. 4.9) and OARSI scores (2.8 vs. 6.0) than untreated mice with ACL-rupture. JQ1 and Flavopiridol work synergistically to reduce injury response after joint trauma, suggesting that targeting Brd4 and/or CDK9 could be a viable strategy for PTOA prevention and treatment of early OA.
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影响因子：
7
作者：
Doyran, B.;Tong, W.;Li, Q.;Jia, H.;Zhang, X.;Chen, C.;Enomoto-Iwamoto, M.;Lu, X. L.;Qin, L.;Han, L.
通讯作者：
Han, L.
影响因子：
--
作者：
Lawrence, Reva C.;Felson, David T.;Wolfe, Frederick
通讯作者：
Wolfe, Frederick
影响因子：
2.8
作者：
Krenn, V;Morawietz, L;König, A
通讯作者：
König, A
影响因子：
7
作者：
Christiansen, B. A.;Anderson, M. J.;Haudenschild, D. R.
通讯作者：
Haudenschild, D. R.
影响因子：
7
作者：
Glasson, S. S.;Blanchet, T. J.;Morris, E. A.
通讯作者：
Morris, E. A.