Chondroitin synthase-3 regulates nucleus pulposus degeneration through actin-induced YAP signaling.
Chondroitin synthase-3 regulates nucleus pulposus degeneration through actin-induced YAP signaling.
复制标题
软骨素合酶 3 通过肌动蛋白 诱导的 YAP 信号调节髓核变性
DOI：
10.1096/fj.202001021r
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Yuan Wen
中科院分区：
文献类型：
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作者：
Wei Leixin;Cao Peng;Xu Chen;Zhong Huajian;Wang Xiukun;Bai Meizhu;Hu Bo;Wang Ruizhe;Liu Ning;Tian Ye;Chen Huajiang;Li Jinsong;Yuan Wen
硫酸软骨素（CS）的丢失已被报道在椎间盘退变（IDD）过程中起关键作用。然而，CS及其糖苷酶的详细机制尚未阐明。由于CS主要由软骨素脱氢酶3（Chsy 3）合成，因此，在此，通过使用CRISPR-Cas9和半克隆技术产生Chsy 3敲除小鼠，以研究其在IDD中的机制。我们发现CS和Chsy 3的表达在IDD过程中在人类和小鼠髓核（NP）组织中均降低，并且敲除Chsy 3表明自发IDD表型与Chsy 3 −/−小鼠中的人类样本相似。利用RNA-Seq数据，我们证实了Chsy 3 −/−NP细胞中分解代谢增加和合成代谢减少的变化。通过生物信息学分析和验证，我们发现Hippo信号通路在Chsy 3 −/−NP细胞中显著下调，并且Yap 1的激活主要受到影响。此外，功能分析表明，Chsy 3可以通过肌动蛋白张力介导的Yap 1激活来调节NP细胞变性，这不依赖于Hippo/Lats信号传导。总之，我们的研究结果揭示了一种新的机制，即CS相关的Chsy 3缺失可通过NP细胞中CS相关的肌动蛋白张力介导雅普激活而引起自发性椎间盘退变。
Loss of chondroitin sulfate (CS) has been reported to play a key role during intervertebral disc degeneration (IDD). However, the detailed mechanism of CS and its synthases have not been elucidated. Since CS is mainly synthesized by chondroitin synthases 3 (Chsy3), here, the Chsy3 knockout mice are generated by using CRISPR‐Cas9 and semi‐cloning technology to study its mechanism during IDD. We find that CS and Chsy3 expression are decreased during IDD both in human and mice nucleus pulposus (NP) tissue, and knockout of Chsy3 shows that spontaneous IDD phenotype resembles that of human samples in theChsy3−/−mice. Taking advantage of RNA‐Seq data, we confirm increased catabolic and decreased anabolic changes inChsy3−/−NP cells. By using bioinformatic analysis and validation, we find that Hippo signaling pathway is significantly downregulated, and the activation of Yap1 is mainly affected inChsy3−/−NP cells. Furthermore, functional analyses have shown that Chsy3 could regulate NP cell degeneration by Actin tension mediated activation of Yap1, which is independent of Hippo/Lats signaling. In summary, our findings reveal a novel mechanism that depletion of CS‐related Chsy3 can cause spontaneous intervertebral disc degeneration by mediating Yap activation through CS‐related actin‐tension in NP cells.