CaMKII inhibition in human primary and pluripotent stem cell-derived chondrocytes modulates effects of TGFβ and BMP through SMAD signaling.
CaMKII inhibition in human primary and pluripotent stem cell-derived chondrocytes modulates effects of TGFβ and BMP through SMAD signaling.
复制标题
DOI：
10.1016/j.joca.2018.08.017
复制
发表时间：
2019-01
影响因子：
7
通讯作者：
Evseenko D
中科院分区：
文献类型：
--
作者：
Saitta B;Elphingstone J;Limfat S;Shkhyan R;Evseenko D
CaMKII 的上调与 OA 的发病机制和关节软骨肥大的重新激活有关。然而，直接抑制 CaMKII 意外地增强了动物模型中的 OA 症状。 CaMKII 在 OA 中的作用尚不清楚，需要进一步研究。在正常人和 OA 关节软骨细胞中进行 CaMKII 表达分析，并使用 CaMKII 的药理学 (KN93)、肽 (AC3-I) 和 siRNA 抑制剂评估关节、胎儿和 PSC 衍生的人软骨细胞中的信号传导机制。磷酸化 CaMKII (pCaMKII) 的表达水平在人类 OA 样本中显着且持续增加。 BMP2/4 激活人成人关节软骨细胞中 pCaMKII 以及 COLII 和 COLX 的表达，并且还增加 SMADs1/5/8 的水平和核定位，而 TGFβ1 显示对成人软骨细胞中软骨形成程序的激活很少或没有。在 BMP4 和 TGFβ1 存在的情况下，用特异性 siRNA 靶向阻断 CaMKII 可降低正常和 OA 成人关节软骨细胞中 pSMAD、COLII、COLX 和蛋白聚糖的水平。在 CaMKII 小分子和肽抑制剂存在下，人类胎儿和 PSC 来源的软骨细胞也显示软骨形成分化减少。此外，SMADs1/5/8 或 2/3 的免疫沉淀随后 pCaMKII 的蛋白质印迹显示原代软骨细胞中 SMADs 和 pCaMKII 之间的直接相互作用。目前的研究表明，CaMKII 在原代软骨细胞和 PSC 衍生软骨细胞中 TGF-β 和 BMP 介导的反应中发挥直接作用。这些发现对干细胞软骨组织的组织工程和骨关节炎的治疗管理具有直接影响。
Upregulation of CaMKII is implicated in the pathogenesis of OA and reactivation of articular cartilage hypertrophy. However, direct inhibition of CaMKII unexpectedly augmented symptoms of OA in animal models. The role of CaMKII in OA remains unclear and requires further investigation. Analysis of CaMKII expression was performed in normal human and OA articular chondrocytes, and signaling mechanisms were assessed in articular, fetal and PSC-derived human chondrocytes using pharmacological (KN93), peptide (AC3-I) and siRNA inhibitors of CaMKII. Expression levels of phospho-CaMKII (pCaMKII) were significantly and consistently increased in human OA specimens. BMP2/4 activated expression of pCaMKII as well as COLII and COLX in human adult articular chondrocytes, and also increased the levels and nuclear localization of SMADs1/5/8, while TGFβ1 showed minimal or no activation of the chondrogenic program in adult chondrocytes. Targeted blockade of CaMKII with specific siRNAs decreased levels of pSMADs, COLII, COLX and proteoglycans in normal and OA adult articular chondrocytes in the presence of both BMP4 and TGFβ1. Both human fetal and PSC-derived chondrocytes also demonstrated a decrease of chondrogenic differentiation in the presence of small molecule and peptide inhibitors of CaMKII. Furthermore, immunoprecipitation for SMADs1/5/8 or 2/3 followed by western blotting for pCaMKII showed direct interaction between SMADs and pCaMKII in primary chondrocytes. Current study demonstrates a direct role for CaMKII in TGF-β and BMP-mediated responses in primary and PSC-derived chondrocytes. These findings have direct implications for tissue engineering of cartilage tissue from stem cells and therapeutic management of OA.
登录
查看更多内容
影响因子：
3.1
作者：
Steward, A. J.;Kelly, D. J.;Wagner, D. R.
通讯作者：
Wagner, D. R.
影响因子：
2.7
作者：
Taschner, Michael J.;Rafigh, Mehran;Hartmann, Christine
通讯作者：
Hartmann, Christine
影响因子：
5.3
作者：
Wicks, SJ;Lui, S;Chantry, A
通讯作者：
Chantry, A
影响因子：
27.4
作者：
Shkhyan R;Van Handel B;Bogdanov J;Lee S;Yu Y;Scheinberg M;Banks NW;Limfat S;Chernostrik A;Franciozi CE;Alam MP;John V;Wu L;Ferguson GB;Nsair A;Petrigliano FA;Vangsness CT;Vadivel K;Bajaj P;Wang L;Liu NQ;Evseenko D
通讯作者：
Evseenko D
影响因子：
4.6
作者：
Li, Yuwei;Ahrens, Molly J.;Dudley, Andrew T.
通讯作者：
Dudley, Andrew T.