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Chaperone-mediated Autophagy Regulates Cell Growth by Targeting SMAD3 in Glioma

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Abstract

Previous studies suggest that the reduction of SMAD3 (mothers against decapentaplegic homolog 3) has a great impact on tumor development, but its exact pathological function remains unclear. In this study, we found that the protein level of SMAD3 was greatly reduced in human-grade IV glioblastoma tissues, in which LAMP2A (lysosome-associated membrane protein type 2A) was significantly up-regulated. LAMP2A is a key rate-limiting protein of chaperone-mediated autophagy (CMA), a lysosome pathway of protein degradation that is activated in glioma. We carefully analyzed the amino-acid sequence of SMAD3 and found that it contained a pentapeptide motif biochemically related to KFERQ, which has been proposed to be a targeting sequence for CMA. In vitro, we confirmed that SMAD3 was degraded in either serum-free or KFERQ motif deleted condition, which was regulated by LAMP2A and interacted with HSC70 (heat shock cognate 71 kDa protein). Using isolated lysosomes, amino-acid residues 75 and 128 of SMAD3 were found to be of importance for this process, which affected the CMA pathway in which SMAD3 was involved. Similarly, down-regulating SMAD3 or up-regulating LAMP2A in cultured glioma cells enhanced their proliferation and invasion. Taken together, these results suggest that excessive activation of CMA regulates glioma cell growth by promoting the degradation of SMAD3. Therefore, targeting the SMAD3–LAMP2A-mediated CMA-lysosome pathway may be a promising approach in anti-cancer therapy.

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Acknowledgments

The authors would like to thank Professors Qian Yang and Pingyi Xu for two years of patient guidance. This work was supported by the National Key R&D Program of China (2016YFC1306601 and 2017YFC1306002), the National Natural Science Foundation of China (82071416, 81870992, 81870856, U1603281, 81903958, 81901282, and 82004459), the Natural Science Foundation of Guangdong Province (2020A1515010986, 2019A1515011189, and 2018A030310521), a Science and Technology Planning Project of Guangdong Province (A2018315), a Technology Project of Guangzhou (2018-1202-SF-0019 and 2019ZD09), and a China Postdoctoral Science Foundation Grant (2019M662873).

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  1. Hanqun Liu, Yuxuan Yong, and Xingjian Li have contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510120, China

    Hanqun Liu, Xingjian Li, Panghai Ye, Guoyou Peng, Mingshu Mo, Wenyuan Guo, Xiang Chen, Yangfu Luo, Yuwan Lin, Zhiling Zhang, Liuyan Ding, Miaomiao Zhou, Lin Lu & Pingyi Xu

  2. Department of Neurosurgery, Tangdu Hospital, The Fourth Military Medical University, Xi’an, 710038, China

    Kai Tao & Jiewen Qiu

  3. Department of Neurology, The Second Affiliated Hospital of Xinjiang Medical University, Ürümqi, 830054, China

    Yuxuan Yong & Xinling Yang

  4. Department of Experiment Surgery, Tangdu Hospital, The Fourth Military Medical University, Xi’an, 710038, China

    Qian Yang

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  1. Hanqun Liu
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Liu, H., Yong, Y., Li, X. et al. Chaperone-mediated Autophagy Regulates Cell Growth by Targeting SMAD3 in Glioma. Neurosci. Bull. 38, 637–651 (2022). https://doi.org/10.1007/s12264-022-00818-9

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