O-GlcNAc modification regulates autophagy and apoptosis in endometriosis

Endometriosis is a common, chronic gynecological disorder characterized by the presence of endometrial-like tissue outside the uterine cavity, frequently associated with significant morbidities such as pelvic pain and infertility. Elucidating its underlying pathogenic mechanisms is therefore of critical importance. O-GlcNAcylation, a ubiquitous post-translational modification, plays pivotal roles in diverse biological processes, yet its involvement in endometriosis progression remains largely unexplored. Here, we demonstrate that aberrantly elevated O-GlcNAcylation contributes to endometriosis pathogenesis by modulating autophagy and apoptosis. Specifically, O-GlcNAcylation levels were elevated in both ectopic and eutopic endometrial tissues, coinciding with compromised autophagic function. In 12Z endometriotic cells, the targeted downregulation of O-GlcNAcylation inhibited cell proliferation while promoting autophagy and apoptosis, whereas its upregulation yielded opposite effects. Furthermore, concurrent inhibition of autophagy reversed the pro-apoptotic effects induced by reduced O-GlcNAcylation. Mechanistically, attenuating O-GlcNAcylation promoted autophagy and apoptosis via the inhibition of the mTOR signaling pathway, an effect potentially mediated by decreased O-GlcNAcylation of Raptor, a core mTORC1 component. In vivo studies further corroborated that suppressing O-GlcNAcylation inhibits ectopic lesion growth and enhances both autophagy and apoptosis. Collectively, our findings reveal that O-GlcNAcylation drives endometriosis progression by regulating autophagy and apoptosis via the mTOR pathway, providing novel mechanistic insights and highlighting potential therapeutic targets for precision interventions. Similar content being viewed by others Data availability The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.

Allaire C, Bedaiwy MA, Yong PJ (2023) Diagnosis and management of endometriosis. CMAJ 195(10):E363–E371. https://doi.org/10.1503/cmaj.220637 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):1244–51e1. https://doi.org/10.1016/j.fertnstert.2015.02.007 Caldwell SA, Jackson SR, Shahriari KS, Lynch TP, Sethi G, Walker S, Vosseller K, Reginato MJ (2010) Nutrient sensor O-GlcNAc transferase regulates breast cancer tumorigenesis through targeting of the oncogenic transcription factor FoxM1. Oncogene 29(19):2831–2842. https://doi.org/10.1038/onc.2010.41 Chen J, Du H, Tong R, Yang H, Ma H (2015) Effect of Bushenwenyanghuayu decoction on nerve growth factor and bradykinin/bradykinin B1 receptor in a endometriosis dysmenorrhea mouse model. Journal Traditional Chin Med 35:184–191. https://doi.org/10.1016/s0254-6272(15)30026-1 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. https://doi.org/10.1093/molehr/gat091 Choi J, Jo M, Lee E, Lee DY, Choi D (2015) Dienogest enhances autophagy induction in endometriotic cells by impairing activation of AKT, ERK1/2, and mTOR. Fertil Steril 104(3):655–64e1. https://doi.org/10.1016/j.fertnstert.2015.05.020 Guo Z, Yu Q (2019) Role of mTOR Signaling in Female Reproduction. Front Endocrinol (Lausanne) 10:692. https://doi.org/10.3389/fendo.2019.00692 Han X, Li X, Liu H, Zhang H, Li A, Dong M, Xu Y, Yan B, Sui L, Kong Y (2019) O-GlcNAc modification influences endometrial receptivity by promoting endometrial cell proliferation, migration and invasion. Oncol Rep 42(5):2065–2074. https://doi.org/10.3892/or.2019.7317 Hanover JA, Chen W, Bond MR (2018) O-GlcNAc in cancer: An Oncometabolism-fueled vicious cycle. J Bioenerg Biomembr 50(3):155–173. https://doi.org/10.1007/s10863-018-9751-2 Hu Y, You C, Song C, Shi Y, Ye L (2022) The beneficial effect of global O-GlcNAcylation on odontogenic differentiation of human dental pulp cells via mTORC1 pathway. Arch Oral Biol 138:105427. https://doi.org/10.1016/j.archoralbio.2022.105427 Kim JJ, Kurita T, Bulun SE (2013) Progesterone action in endometrial cancer, endometriosis, uterine fibroids, and breast cancer. Endocr Rev 34(1):130–162. https://doi.org/10.1210/er.2012-1043 Kim J, Kundu M, Viollet B, Guan KL (2019) AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol 13(2):132–141. https://doi.org/10.1038/ncb2152 Kim BS, Kim B, Yoon S, Park W, Bae SJ, Joo J, Kim W, Ha KT (2025) Warburg-like Metabolic Reprogramming in Endometriosis: From Molecular Mechanisms to Therapeutic Approaches. Pharmaceuticals (Basel) 18(6):813. https://doi.org/10.3390/ph18060813 Leconte M, Nicco C, Ngô C, Chéreau C, Chouzenoux S, Marut W, Guibourdenche J, Arkwright S, Weill B, Chapron C, Dousset B, Batteux F (2011) The mTOR/AKT inhibitor temsirolimus prevents deep infiltrating endometriosis in mice. Am J Pathol 179(2):880–889. https://doi.org/10.1016/j.ajpath.2011.04.020 McKinnon BD, Kocbek V, Nirgianakis K, Bersinger NA, Mueller MD (2016) Kinase signalling pathways in endometriosis: potential targets for non-hormonal therapeutics. Hum Reprod Update 22(3):382–403. https://doi.org/10.1093/humupd/dmv060 Mei J, Zhu XY, Jin LP, Duan ZL, Li DJ, Li MQ (2015) Estrogen promotes the survival of human secretory phase endometrial stromal cells via CXCL12/CXCR4 up-regulation-mediated autophagy inhibition. Hum Reprod 30(7):1677–1689. https://doi.org/10.1093/humrep/dev100 Mizushima N, Levine B (2020) Autophagy in Human Diseases. N Engl J Med 383(16):1564–1576. https://doi.org/10.1056/NEJMra2022774 Murakami K, Kurotaki D, Kawase W, Soma S, Fukuchi Y, Kunimoto H, Yoshimi R, Koide S, Oshima M, Hishiki T, Hayakawa N, Matsuura T, Oda M, Yanagisawa K, Kobayashi H, Haraguchi M, Atobe Y, Funakoshi K, Iwama A, Takubo K, Okamoto S, Tamura T, Nakajima H (2020) OGT Regulates Hematopoietic Stem Cell Maintenance via PINK1-Dependent Mitophagy. Cell Rep 34(1):108579. https://doi.org/10.1016/j.celrep.2020.108579 Ruiz A, Rockfield S, Taran N, Haller E, Engelman RW, Flores I, Panina-Bordignon P, Nanjundan M (2016) Effect of hydroxychloroquine and characterization of autophagy in a mouse model of endometriosis. Cell Death Dis 7(1):e2059. https://doi.org/10.1038/cddis.2015.361 Sampson JA (1927) Metastatic or Embolic Endometriosis, due to the Menstrual Dissemination of Endometrial Tissue into the Venous Circulation. Am J Pathol 3(2):93–11043 Saunders PTK, Horne AW (2021) Endometriosis: Etiology, pathobiology, and therapeutic prospects. Cell 184(11):2807–2824. https://doi.org/10.1016/j.cell.2021.04.041 Shi Y, Tomic J, Wen F, Shaha S, Bahlo A, Harrison R, Dennis JW, Williams R, Gross BJ, Walker S, Zuccolo J, Deans JP, Hart GW, Spaner DE (2010) Aberrant O-GlcNAcylation characterizes chronic lymphocytic leukemia. Leukemia 24(9):1588–1598. https://doi.org/10.1038/leu.2010.152 Taniguchi F, Kaponis A, Izawa M, Kiyama T, Deura I, Ito M, Iwabe T, Adonakis G, Terakawa N, Harada T (2011) Apoptosis and endometriosis. Front Biosci (Elite Ed) 3(2):648–662. https://doi.org/10.2741/e277 Taylor HS, Kotlyar AM, Flores VA (2021) Endometriosis is a chronic systemic disease: clinical challenges and novel innovations. Lancet 397(10276):839–852. https://doi.org/10.1016/S0140-6736(21)00389-5 Yang HL, Zhou WJ, Chang KK, Mei J, Huang LQ, Wang MY, Meng Y, Ha SY, Li DJ, Li MQ (2017) The crosstalk between endometrial stromal cells and macrophages impairs cytotoxicity of NK cells in endometriosis by secreting IL-10 and TGF-β. Reproduction 154(6):815–825. https://doi.org/10.1530/REP-17-0342 Yao R, Yang Y, Lian S, Shi H, Liu P, Liu Y, Yang H, Li S (2018) Effects of Acute Cold Stress on Liver O-GlcNAcylation and Glycometabolism in Mice. Int J Mol Sci 19(9):2815. https://doi.org/10.3390/ijms19092815 Zachara NE (2012) The roles of O-linked β-N-acetylglucosamine in cardiovascular physiology and disease. Am J Physiol Heart Circ Physiol 15(10):H1905–H1918. https://doi.org/10.1152/ajpheart.00445.2011 Zhai L, Yang X, Dong J, Qian L, Gao Y, Lv Y, Chen L, Chen B, Zhou F (2023) O–GlcNAcylation mediates endometrial cancer progression by regulating the Hippo–YAP pathway. Int J Oncol 63(2):90. https://doi.org/10.3892/ijo.2023.5538 Zhang H, Qi J, Pei J, Zhang M, Shang Y, Li Z, Wang Y, Guo J, Sun K, Fan J, Sui L, Xu Y, Kong L, Kong Y (2021) O-GlcNAc modification mediates aquaporin 3 to coordinate endometrial cell glycolysis and affects embryo implantation. J Adv Res 37:119–131. https://doi.org/10.1016/j.jare.2021.06.022 Zhang M, Xu T, Tong D, Li S, Yu X, Liu B, Jiang L, Liu K (2023) Research advances in endometriosis-related signaling pathways: A review. Biomed Pharmacother 164:114909. https://doi.org/10.1016/j.biopha.2023.114909 Zhou F, Yang X, Zhao H, Liu Y, Feng Y, An R, Lv X, Li J, Chen B (2018) Down-regulation of OGT promotes cisplatin resistance by inducing autophagy in ovarian cancer. Theranostics 8(19):5200–5212. https://doi.org/10.7150/thno.27806 Zhou Y, Peng Y, Xia Q, Yan D, Zhang H, Zhang L, Chen Y, Zhao X, Li J (2021) Decreased Indian hedgehog signaling activates autophagy in endometriosis and adenomyosis. Reproduction 161(2):99–109. https://doi.org/10.1530/REP-20-0172

We express our gratitude to H.S. Zhang for his help in Experimental Approach Guidance.We also gratefully acknowledge figdraw for their support in creating the flowchart in Fig. 2A. Funding This work was supported by the Key Research and Development Program of Liaoning Province, China (Grant No. 2022JH1/10800070). No other funding was received. Author information Authors and Affiliations Contributions X.H. Jin, Y.L. Feng and L. Xu conceived and designed the study. X.H. Jin and L.P. Fan performed the experiments and wrote the manuscript. J. Li, J.G. Li and R.Y. Liu contributed to data analysis. Y.H. Shang, Y. Kong, and Q. Yang revised the manuscript. All the authors contributed to the interpretation of the data and the revision of the manuscript and helped at various stages of the study. All authors read and approved the final manuscript. Corresponding authors Ethics declarations Competing interests The authors declare no competing interests. Additional information Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Supplementary Information Below is the link to the electronic supplementary material. 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 Jin, X., Feng, Y., Xu, L. et al. O-GlcNAc modification regulates autophagy and apoptosis in endometriosis. Funct Integr Genomics 26, 109 (2026). https://doi.org/10.1007/s10142-026-01888-y Received: Revised: Accepted: Published: Version of record: DOI: https://doi.org/10.1007/s10142-026-01888-y