Transcription factor EB-mediated autophagy affects cell migration and inhibits apoptosis to promote endometriosis

Autophagy has emerged as an important process of cell metabolism. With continuous in-depth research on autophagy, TFEB has been a key transcription factor regulating autophagy levels in recent years. Studies have established that TFEB regulates autophagy and apoptosis in various diseases. However, the relationship between TFEB and the pathogenesis of endometriosis remains unclear. This study aimed to investigate the effect of TFEB on the mechanism of endometriosis progression. The results showed that TFEB and autophagy-related protein LC3 are highly expressed in ectopic endometrium of patients with endometriosis, overexpression of TFEB in cultured human endometrial stromal cells (HESCs) by lentivirus not only promoted autophagy but also inhibited apoptosis. In addition, the migration and invasion ability of HESCs were enhanced by TFEB overexpression. Furthermore, inhibiting autophagy with specific inhibitors can attenuate migration and invasion of HESCs induced by TFEB. The rat models of endometriosis show that TFEB knockdown can suppress lesion growth in vivo. Our results suggest that autophagy may be involved in the progression mechanism of endometriosis, and the mechanism of autophagy disorder in endometriosis is probably related to TFEB. TFEB may be a key molecule in promoting endometriosis. Similar content being viewed by others Data availability The data that support the findings of this study are available on resquest from the corresponding author. Abbreviations - CQ: - Chloroquine - HESCs: - Human endometrial stromal cells - mTORC1: - Mammalian target of rapamycin complex 1 - TFEB: - Transcription factor EB

Liu Y, Wang J, Zhang X (2022) An update on the multifaceted role of NF-kappaB in endometriosis. Int J Biol Sci 18(11):4400–4413 Epub 2022/07/23. https://doi.org/10.7150/ijbs.72707 Kvaskoff M, Mu F, Terry KL, Harris HR, Poole EM, Farland L et al (2015) Endometriosis: a high-risk population for major chronic diseases? Hum Reprod Update 21(4):500–516 Epub 2015/03/15. https://doi.org/10.1093/humupd/dmv013 Lagana AS, Garzon S, Gotte M, Vigano P, Franchi M, Ghezzi F et al (2019) The pathogenesis of endometriosis: molecular and cell biology insights. Int J Mol Sci 20(22). Epub 2019/11/14. https://doi.org/10.3390/ijms20225615. PubMed PMID: 31717614; PubMed Central PMCID: PMCPMC6888544 Li X, He S, Ma B (2020) Autophagy and autophagy-related proteins in cancer. Mol Cancer 19(1):12 Epub 2020/01/24. https://doi.org/10.1186/s12943-020-1138-4 Shibutani ST, Saitoh T, Nowag H, Munz C, Yoshimori T (2015) Autophagy and autophagy-related proteins in the immune system. Nat Immunol 16(10):1014–1024 Epub 2015/09/19. https://doi.org/10.1038/ni.3273 Liu H, Zhang Z, Xiong W, Zhang L, Xiong Y, Li N et al (2017) Hypoxia-inducible factor-1alpha promotes endometrial stromal cells migration and invasion by upregulating autophagy in endometriosis. Reproduction 153(6):809–820 Epub 2017/03/30. https://doi.org/10.1530/REP-16-0643 Allavena G, Carrarelli P, Del Bello B, Luisi S, Petraglia F, Maellaro E (2015) Autophagy is upregulated in ovarian endometriosis: a possible interplay with p53 and heme oxygenase-1. Fertil Steril 103(5):1244–1251 .007. PubMed PMID: 25772769 Settembre C, Di Malta C, Polito VA, Garcia Arencibia M, Vetrini F, Erdin S et al (2011) TFEB links autophagy to lysosomal biogenesis. Science 332(6036):1429–1433 Epub 2011/05/28. https://doi.org/10.1126/science.1204592 Napolitano G, Ballabio A (2016) TFEB at a glance. J Cell Sci 129(13):2475–2481 Epub 2016/06/03. 10.1242 Chen M, Dai Y, Liu S, Fan Y, Ding Z, Li D (2021) TFEB biology and agonists at a glance. Cells 10(2). https://doi.org/10.3390/cells10020333PubMed PMID: 33562649; PubMed Central PMCID: PMCPMC7914707 Epub 2021/02/11 Cortes CJ, La Spada AR (2019) TFEB dysregulation as a driver of autophagy dysfunction in neurodegenerative disease: molecular mechanisms, cellular processes, and emerging therapeutic opportunities. Neurobiol Dis 122:83–93 Epub 2018/06/01. https://doi.org/10.1016/j.nbd.2018.05.012 Calio A, Brunelli M, Segala D, Pedron S, Remo A, Ammendola S et al (2020) Comprehensive analysis of 34 MiT family translocation renal cell carcinomas and review of the literature: investigating prognostic markers and therapy targets. Pathology 52(3):297–309 PubMed PMID: 32107074 Kuiper RP, Schepens M, Thijssen J, Schoenmakers EF, van Kessel AG (2004) Regulation of the MiTF/TFE bHLH-LZ transcription factors through restricted spatial expression and alternative splicing of functional domains. Nucleic Acids Res 32(8):2315–2322 Epub 2004/05/01. https://doi.org/10.1093/nar/gkh571 Marino G, Niso-Santano M, Baehrecke EH, Kroemer G (2014) Self-consumption: the interplay of autophagy and apoptosis. Nat Rev Mol Cell Biol 15(2):81–94 Epub 2014/01/10. https://doi.org/10.1038/nrm3735 Zheng G, Pan Z, Zhan Y, Tang Q, Zheng F, Zhou Y et al (2019) TFEB protects nucleus pulposus cells against apoptosis and senescence via restoring autophagic flux. Osteoarthritis Cartilage 27(2):347–357 Epub 2018/11/12. https://doi.org/10.1016/j.joca.2018.10.011 Mizushima N, Yoshimori T (2007) How to interpret LC3 immunoblotting. Autophagy 3(6):542–5. Epub 2007/07/06. https://doi.org/10.4161/auto.4600. PubMed PMID: 17611390 Sun X, Chang R, Tang Y, Luo S, Jiang C, Jia H et al (2021) Transcription factor EB (TFEB)-mediated autophagy protects bovine mammary epithelial cells against H(2)O(2)-induced oxidative damage in vitro. J Anim Sci Biotechnol 12(1):35. https://doi.org/10.1186/s40104-021-00561-7PubMed PMID: 33685494; PubMed Central PMCID: PMCPMC7941962 Epub 2021/03/10 Vercellini P, Vigano P, Somigliana E, Fedele L (2014) Endometriosis: pathogenesis and treatment. Nat Rev Endocrinol 10(5):261–275 Epub 2013/12/25. https://doi.org/10.1038/nrendo.2013.255 Iwabuchi T, Yoshimoto C, Shigetomi H, Kobayashi H (2015) Oxidative stress and antioxidant defense in endometriosis and its malignant transformation. Oxid Med Cell Longev 2015:848595. Epub 2015/07/18 https://doi.org/10.1155/2015/848595 White E (2015) The role for autophagy in cancer. J Clin Invest 125(1):42–46 Epub 2015/02/06. doi: 10.1172/JCI73941. PubMed PMID: 25654549; PubMed Central PMCID: PMCPMC4382247 Stamatakou E, Wrobel L, Hill SM, Puri C, Son SM, Fujimaki M et al (2020) Erratum: author correction: mendelian neurodegenerative disease genes involved in autophagy. Cell Discov 6:40 Epub 2020/06/23. https://doi.org/10.1038/s41421-020-0185-8 Lin XX, Sen I, Janssens GE, Zhou X, Fonslow BR, Edgar D et al (2018) DAF-16/FOXO and HLH-30/TFEB function as combinatorial transcription factors to promote stress resistance and longevity. Nat Commun 9(1):4400. https://doi.org/10.1038/s41467-018-06624-0PubMed PMID: 30353013; PubMed Central PMCID: PMCPMC6199276 Epub 2018/10/26 Ma S, Fang Z, Luo W, Yang Y, Wang C, Zhang Q et al (2016) The C-ETS2-TFEB axis promotes neuron survival under oxidative stress by regulating lysosome activity. Oxid Med Cell Longev 2016:4693703. Epub 2016/05/20 https://doi.org/10.1155/2016/4693703 Zhitomirsky B, Yunaev A, Kreiserman R, Kaplan A, Stark M, Assaraf YG (2018) Lysosomotropic drugs activate TFEB via lysosomal membrane fluidization and consequent inhibition of mTORC1 activity. Cell Death Dis 9(12):1191 Epub 2018/12/14. https://doi.org/10.1038/s41419-018-1227-0 Fang S, Wan X, Zou X, Sun S, Hao X, Liang C et al (2021) Arsenic trioxide induces macrophage autophagy and atheroprotection by regulating ROS-dependent TFEB nuclear translocation and AKT/mTOR pathway. Cell Death Dis 12(1):88 Epub 2021/01/20. https://doi.org/10.1038/s41419-020-03357-1 Ding Y, Zhu Q, He Y, Lu Y, Wang Y, Qi J et al (2021) Induction of autophagy by beclin-1 in granulosa cells contributes to follicular progesterone elevation in ovarian endometriosis. Transl Res 227:15–29 Epub 2020/07/09. https://doi.org/10.1016/j.trsl.2020.06.013 Zhu L, Yuan Y, Yuan L, Li L, Liu F, Liu J et al (2020) Activation of TFEB-mediated autophagy by trehalose attenuates mitochondrial dysfunction in cisplatin-induced acute kidney injury. Theranostics 10(13):5829–5844 Epub 2020/06/03. https://doi.org/10.7150/thno.44051 Choi J, Jo M, Lee E, Kim HJ, Choi D (2014) Differential induction of autophagy by mTOR is associated with abnormal apoptosis in ovarian endometriotic cysts. Mol Hum Reprod 20(4):309–317 Epub 2013/12/10. https://doi.org/10.1093/molehr/gat091 Zhang X, Cheng X, Yu L, Yang J, Calvo R, Patnaik S et al (2016) MCOLN1 is a ROS sensor in lysosomes that regulates autophagy. Nat Commun 7:12109 Epub 2016/07/01. https://doi.org/10.1038/ncomms12109 Wang H, Wang N, Xu D, Ma Q, Chen Y, Xu S et al (2020) Oxidation of multiple MiT/TFE transcription factors links oxidative stress to transcriptional control of autophagy and lysosome biogenesis. Autophagy 16(9):1683–1696 PubMed PMID: 31826695; PubMed Central PMCID: PMCPMC8386635 Chen Y, Henson ES, Xiao W, Huang D, McMillan-Ward EM, Israels SJ et al (2016) Tyrosine kinase receptor EGFR regulates the switch in cancer cells between cell survival and cell death induced by autophagy in hypoxia. Autophagy 12(6):1029–1046 PubMed PMID: 27166522; PubMed Central PMCID: PMCPMC4922445 Zhan HQ, Qin R, Li YL, Liu MM, Gan L (2021) TFEB promotes BCL-2 expression by upregulating its promoter activity in the t(6;11) translocation renal cell carcinomas. Am J Transl Res 13(8):8804–8818 Epub 2021/09/21. PubMed PMID: 34539996; PubMed Central PMCID: PMCPMC8430107 Xie C, Shi Y, Chen Z, Zhou X, Luo P, Hong C et al (2021) Apigenin alleviates intervertebral disc degeneration via restoring autophagy flux in nucleus pulposus cells. Front Cell Dev Biol 9:787278 Epub 2022/02/01. https://doi.org/10.3389/fcell.2021.787278

The authors thank the staff of the Second Affiliated Hospital of Wenzhou Medical. Funding This study was financially supported by the Natural Science Foundation of Zhejiang Province [grant No. LY20H040005]. The Public Welfare Science and Technology Project of Wenzhou City [grant No. Y20210021]. The Medical Science and Technology Project of Zhejiang Province [grant No.2023KY145]. The Natural Science Foundation of Zhejiang Province [grant No. LY23H040003]. The Medical Science and Technology Project of Zhejiang Province [grant No. 2022KY212]. The Zhejiang Province Traditional Chinese Medicine Science and Technology Project [grantNo.2023ZL510]. The Public Welfare Science and Technology Project of Wenzhou City [grant No. Y2020087]. Author information Authors and Affiliations Contributions Zhao Yu and Qiong Zhang designed the experiments and revised manuscript; Chen Qiuyu and Sennan Zhu conducted experiments and wrote the main manuscript; Yu Meng Qi and Zhou Yi analysed data; Sun Jindan and Du Wenzhuo prepared Figs. 1 and 2; Chen Ziqi, Tao Jiayu and Feng Xiao prepared 3, 4, 5, 6 and 7. Corresponding authors Ethics declarations Statements and declarations The research results reported in this article are all original research results of myself or all members of our research group. Competing interests The authors declare no competing interests. Ethical approval This study was approved by the Ethics Committee of Wenzhou Medical University Affiliated Second Hospital and Yuying Children’s Hospital. (Ethical code LCKY209-132) Additional information Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Rights and permissions Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. About this article Cite this article Chen, Q., Zhou, Y., Yu, M. et al. Transcription factor EB-mediated autophagy affects cell migration and inhibits apoptosis to promote endometriosis. Apoptosis 29, 757–767 (2024). https://doi.org/10.1007/s10495-024-01939-4 Accepted: Published: Version of record: Issue date: DOI: https://doi.org/10.1007/s10495-024-01939-4