MicroRNA-22 promoted osteogenic differentiation of valvular interstitial cells by inhibiting CAB39 expression during aortic valve calcification
MicroRNA-22 promoted osteogenic differentiation of valvular interstitial cells by inhibiting CAB39 expression during aortic valve calcification
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MicroRNA-22通过抑制主动脉瓣钙化过程中CAB39的表达促进瓣膜间质细胞的成骨分化
DOI：
10.1007/s00018-022-04177-6
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发表时间：
2022-03-01
影响因子：
8
通讯作者：
Wang, Guokun
中科院分区：
文献类型：
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作者：
Yang, Fan;Liu, Suxuan;Wang, Guokun
钙化性主动脉瓣疾病（CAVD）是一种常见的瓣膜疾病，其特征是主动脉瓣的纤维钙化重构，这是一个涉及瓣膜间质细胞（VIC）成骨分化的主动调节过程。microRNA（miRNA）是细胞内多种生物学过程的重要调节因子。本研究旨在探讨miR-22在VIC成骨分化中的作用及其机制。在CAVD患者（n= 33）和健康对照（n= 12）的主动脉瓣组织中首次检测到成骨相关miRNA的表达谱。miR-22在钙化瓣膜组织中高表达（P< 0.01），其表达与OPN（rs= 0.820，P < 0.01）和Runx 2（rs= 0.563，P < 0.01）的表达呈正相关。miR-22的持续高表达也在体外VIC成骨模型中得到验证。然后进行腺病毒介导的功能获得和功能丧失实验。miR-22的过表达显著加速了VIC的钙化过程，表现为钙沉积、碱性磷酸酶活性和成骨细胞分化标志物表达的显著增加。相反，miR-22的抑制显著否定了钙化过程。随后，钙结合蛋白39（CAB 39）被鉴定为miR-22的靶点。miR-22的过表达显著降低了VIC中CAB 39的表达，导致CAB 39-LKB 1-STRAD复合物的催化活性降低，这反过来又加剧了AMPK-mTOR信号通路的变化，并最终加速了钙化过程。此外，维克钙化过程中ROS的产生和自噬活性也受到miR-22/CAB 39通路的调控。这些结果表明，miR-22是VIC成骨分化的重要促进剂，并且是CAVD的潜在治疗靶点。
Calcific aortic valve disease (CAVD) is a common valve disease characterized by the fibro-calcific remodeling of the aortic valves, which is an actively regulated process involving osteogenic differentiation of valvular interstitial cells (VICs). MicroRNA (miRNA) is an essential regulator in diverse biological processes in cells. The present study aimed to explore the role and mechanism of miR-22 in the osteogenic differentiation of VICs. The expression profile of osteogenesis-related miRNAs was first detected in aortic valve tissue from CAVD patients (n= 33) and healthy controls (n= 12). miR-22 was highly expressed in calcified valve tissues (P< 0.01), and the expression was positively correlated with the expression of OPN (rs= 0.820,P< 0.01) and Runx2 (rs= 0.563,P< 0.01) in VICs isolated from mild or moderately calcified valves. The sustained high expression of miR-22 was also validated in an in-vitro VICs osteogenic model. Adenovirus-mediated gain-of-function and loss-of-function experiments were then performed. Overexpression of miR-22 significantly accelerated the calcification process of VICs, manifested by significant increases in calcium deposition, alkaline phosphate activity, and expression of osteoblastic differentiation markers. Conversely, inhibition of miR-22 significantly negated the calcification process. Subsequently, calcium-binding protein 39 (CAB39) was identified as a target of miR-22. Overexpression of miR-22 significantly reduced the expression of CAB39 in VICs, leading to decreased catalytic activity of the CAB39–LKB1–STRAD complex, which, in turn, exacerbated changes in the AMPK–mTOR signaling pathway, and ultimately accelerated the calcification process. In addition, ROS generation and autophagic activity during VIC calcification were also regulated by miR-22/CAB39 pathway. These results indicate that miR-22 is an important accelerator of the osteogenic differentiation of VICs, and a potential therapeutic target in CAVD.