Wang Xiaofeng, Qi Huabing, Wang Quan, Zhu Ying, Wang Xianxing, Jin Min, Tan Qiaoyan, Huang Qizhao, Xu Wei, Li Xiaogang, Kuang Liang, Tang Yubing, Du Xiaolan, Chen Di, Chen Lin
a Center of Bone Metabolism and Repair (CBMR); Trauma Center; Institute of Surgery Research; Daping Hospital; Third Military Medical University ; Chongqing , China.
b State Key Laboratory of Trauma; Burns and Combined Injury; Third Military Medical University ; Chongqing , China.
Autophagy. 2015 Nov 2;11(11):1998-2013. doi: 10.1080/15548627.2015.1091551.
FGFR3 (fibroblast growth factor receptor 3) is a negative regulator of endochondral ossification. Gain-of-function mutations in FGFR3 are responsible for achondroplasia, the most common genetic form of dwarfism in humans. Autophagy, an evolutionarily conserved catabolic process, maintains chondrocyte viability in the growth plate under stress conditions, such as hypoxia and nutritional deficiencies. However, the role of autophagy and its underlying molecular mechanisms in achondroplasia remain elusive. In this study, we found activated FGFR3 signaling inhibited autophagic activity in chondrocytes, both in vivo and in vitro. By employing an embryonic bone culture system, we demonstrated that treatment with autophagy inhibitor 3-MA or chloroquine led to cartilage growth retardation, which mimics the effect of activated-FGFR3 signaling on chondrogenesis. Furthermore, we found that FGFR3 interacted with ATG12-ATG5 conjugate by binding to ATG5. More intriguingly, FGFR3 signaling was found to decrease the protein level of ATG12-ATG5 conjugate. Consistently, using in vitro chondrogenic differentiation assay system, we showed that the ATG12-ATG5 conjugate was essential for the viability and differentiation of chondrocytes. Transient transfection of ATG5 partially rescued FGFR3-mediated inhibition on chondrocyte viability and differentiation. Our findings reveal that FGFR3 inhibits the autophagic activity by decreasing the ATG12-ATG5 conjugate level, which may play an essential role in the pathogenesis of achondroplasia.
成纤维细胞生长因子受体3(FGFR3)是软骨内骨化的负调节因子。FGFR3的功能获得性突变是导致软骨发育不全的原因,软骨发育不全是人类最常见的侏儒症遗传形式。自噬是一种进化上保守的分解代谢过程,在缺氧和营养缺乏等应激条件下维持生长板中软骨细胞的活力。然而,自噬在软骨发育不全中的作用及其潜在分子机制仍不清楚。在本研究中,我们发现激活的FGFR3信号在体内和体外均抑制软骨细胞的自噬活性。通过采用胚胎骨培养系统,我们证明用自噬抑制剂3-MA或氯喹处理会导致软骨生长迟缓,这模拟了激活的FGFR3信号对软骨形成的影响。此外,我们发现FGFR3通过与ATG5结合与ATG12-ATG5共轭体相互作用。更有趣的是,发现FGFR3信号降低了ATG12-ATG5共轭体的蛋白质水平。一致地,使用体外软骨形成分化测定系统,我们表明ATG12-ATG5共轭体对于软骨细胞的活力和分化至关重要。ATG5的瞬时转染部分挽救了FGFR3介导的对软骨细胞活力和分化的抑制。我们的研究结果表明,FGFR3通过降低ATG12-ATG5共轭体水平来抑制自噬活性,这可能在软骨发育不全的发病机制中起重要作用。