{"paper_id":"be667e30-32f8-4a0f-b5db-816772209e12","body_text":"Abstract\nEndometriosis is a prevalent gynecological disorder worldwide that significantly impairs the quality of life of patients and imposes a substantial economic burden. The limited efficacy and adverse effects of current pharmacological and surgical treatments highlight the urgent need for alternative diagnostic biomarkers and therapeutic targets. Recent evidence indicates that metabolic reprogramming, particularly lactate metabolism, plays a crucial role in the development and progression of endometriosis. In this study, we employed differential expression analysis to identify differentially expressed genes in endometriosis patients, and used Least Absolute Shrinkage and Selection Operator (LASSO) regression and Random Forest (RF) for feature selection and model construction. Five key genes (MRPS2, YARS2, SLC3A1, CALD1, and S100A4) were validated as highly accurate diagnostic markers. Furthermore, we conducted in-depth analysis of the regulatory activity of these key lactate metabolism-related genes and their association with immune infiltration. Additionally, we constructed a lncRNA-miRNA gene regulatory network, providing insights into the molecular mechanisms and potential therapeutic strategies. Among these key lactate metabolism-related genes, S100A4 was found to be significantly upregulated in the lesions of endometriosis and plays a pivotal role in regulating cellular invasion and metabolic reprogramming in endometriosis. Silencing of S100A4 significantly inhibited the migration and invasion ability of endometrial stromal cells, whereas overexpression of S100A4 reversed this inhibitory effect. This finding further underscores the importance of S100A4 as a potential therapeutic target. In conclusion, this study highlights the potential value of lactate-related genes as diagnostic biomarkers and therapeutic targets, especially targeting the S100A4 gene in endometrial stromal cells, which may serve as an effective therapeutic strategy to inhibit the progression of endometriosis, offering new directions for future clinical diagnosis and treatment.\nSimilar content being viewed by others\nData Availability\nAll data are available in the article or its supporting information, further inquiries can be directed to the corresponding authors.\nReferences\nZondervan KT, Becker CM, Koga K, Missmer SA, Taylor RN, Viganò P. Endometriosis. Nat Rev Dis Primers. 2018;4:9.\nTaylor HS, Kotlyar AM, Flores VA. Endometriosis is a chronic systemic disease: clinical challenges and novel innovations. Lancet. 2021;397:839–52.\nVercellini P, Viganò P, Somigliana E, Fedele L. Endometriosis: pathogenesis and treatment. Nat Rev Endocrinol. 2014;10:261-75.\nCho MK, Jin L, Han JH, Jin JS, Cheon SY, Shin S, Bae SJ, Park JK, Ha KT. Water-extracted prunella vulgaris alleviates endometriosis by reducing aerobic glycolysis. Front Pharmacol. 2022;13: 872810.\nBulun SE. Endometriosis caused by retrograde menstruation: now demonstrated by DNA evidence. Fertil Steril. 2022;118:535-536.\nD’Hooghe TM, Debrock S. Endometriosis, retrograde menstruation and peritoneal inflammation in women and in baboons. Hum Reprod Update. 2002;8:84–8.\nGuo SW, Habiba M, Benagiano G. From retrograde menstruation to endometrial determinism and a brave new world of \"root treatment\" of endometriosis: destiny or a fanciful utopia? Biomolecules. 2023;13(2):336.\nQi X, Zhang Y, Ji H, Wu X, Wang F, Xie M, Shu L, Jiang S, Mao Y, Cui Y, Liu J. Knockdown of prohibitin expression promotes glucose metabolism in eutopic endometrial stromal cells from women with endometriosis. Reprod Biomed Online. 2014;29:761–70.\nLiang Y, Wu J, Wang W, Xie H, Yao S. Pro-endometriotic niche in endometriosis. Reprod Biomed Online. 2019;38:549–59.\nWang Y, Nicholes K, Shih IM. The origin and pathogenesis of endometriosis. Annu Rev Pathol. 2020;15:71–95.\nLu Q, Huang Y, Wu J, Guan Y, Du M, Wang F, Liu Z, Zhu Y, Gong G, Hou H, Zhang M, Zhang JY, Ning F, Chen L, Wang L, Lash GE. T-cadherin inhibits invasion and migration of endometrial stromal cells in endometriosis. Hum Reprod. 2020;35:145–56.\nLiu H, Zhang G, Gao R. Cellular and molecular characteristics of the premetastatic niches. Animal Model Exp Med. 2023;6:399–408.\nWang T, Ye Z, Li Z, Jing DS, Fan GX, Liu MQ, Zhuo QF, Ji SR, Yu XJ, Xu XW, Qin Y. Lactate-induced protein lactylation: A bridge between epigenetics and metabolic reprogramming in cancer. Cell Prolif. 2023;56: e13478.\nWang X, Fan W, Li N, Ma Y, Yao M, Wang G, He S, Li W, Tan J, Lu Q, Hou S. YY1 lactylation in microglia promotes angiogenesis through transcription activation-mediated upregulation of FGF2. Genome Biol. 2023;24:87.\nZhang D, Tang Z, Huang H, Zhou G, Cui C, Weng Y, Liu W, Kim S, Lee S, Perez-Neut M, Ding J, Czyz D, Hu R, Ye Z, He M, Zheng YG, Shuman HA, Dai L, Ren B, Roeder RG, Becker L, Zhao Y. Metabolic regulation of gene expression by histone lactylation. Nature. 2019;574:575–80.\nLi X, Yang Y, Zhang B, Lin X, Fu X, An Y, Zou Y, Wang JX, Wang Z, Yu T. Lactate metabolism in human health and disease. Signal Transduct Target Ther. 2022;7:305.\nLi WN, Wu MH, Tsai SJ. Hypoxia and reproductive health: The role of hypoxia in the development and progression of endometriosis. Reproduction. 2021;161:F19-f31.\nLu C, Qiao P, Fu R, Wang Y, Lu J, Ling X, Liu L, Sun Y, Ren C, Yu Z. Phosphorylation of PFKFB4 by PIM2 promotes anaerobic glycolysis and cell proliferation in endometriosis. Cell Death Dis. 2022;13:790.\nXu YF, Zhao ZB, Yan EP, Lian ZX, Zhang W. Complex interplay between the immune system, metabolism, and epigenetic factors in autoimmune liver diseases. Medicine Advances. 2023;1:97–114.\nGou Y, Wang H, Wang T, Wang H, Wang B, Jiao N, Yu Y, Cao Y, Wang H, Zhang Z. Ectopic endometriotic stromal cells-derived lactate induces M2 macrophage polarization via Mettl3/Trib1/ERK/STAT3 signalling pathway in endometriosis. Immunology. 2023;168:389–402.\nChen J, Qin P, Sun Y, Han S. Histone lactylation promotes cell proliferation, migration and invasion through targeting HMGB1 in endometriosis. J Biomed Res. 2023;37:470–8.\nYoung VJ, Brown JK, Maybin J, Saunders PT, Duncan WC, Horne AW. Transforming growth factor-β induced Warburg-like metabolic reprogramming may underpin the development of peritoneal endometriosis. J Clin Endocrinol Metab. 2014;99:3450–9.\nLi S, Yao J, Zhang S, Zhou X, Zhao X, Di N, Hao S, Zhi H. Prognostic value of tumor-microenvironment-associated genes in ovarian cancer. BIO Integration. 2023;4:84–96.\nSun H, Wang C, Hu B, Gao X, Zou T, Luo Q, Chen M, Fu Y, Sheng Y, Zhang K, Zheng Y, Ren X, Yan S, Geng Y, Yang L, Dong Q, Qin L. Exosomal S100A4 derived from highly metastatic hepatocellular carcinoma cells promotes metastasis by activating STAT3. Signal Transduct Target Ther. 2021;6:187.\nTochimoto M, Oguri Y, Hashimura M, Konno R, Matsumoto T, Yokoi A, Kodera Y, Saegusa M. S100A4/non-muscle myosin II signaling regulates epithelial-mesenchymal transition and stemness in uterine carcinosarcoma. Lab Invest. 2020;100:682–95.\nSchneider M, Hansen JL, Sheikh SP. S100A4: a common mediator of epithelial-mesenchymal transition, fibrosis and regeneration in diseases? J Mol Med (Berl). 2008;86:507–22.\nMchugh J. S100A4 inhibition targets fibrosis in SSc. Nat Rev Rheumatol. 2024;20:67.\nMatsuzaki S, Darcha C. Epithelial to mesenchymal transition-like and mesenchymal to epithelial transition-like processes might be involved in the pathogenesis of pelvic endometriosis. Hum Reprod. 2012;27:712–21.\nHapangama DK, Raju RS, Valentijn AJ, Barraclough D, Hart A, Turner MA, Platt-Higgins A, Barraclough R, Rudland PS. Aberrant expression of metastasis-inducing proteins in ectopic and matched eutopic endometrium of women with endometriosis: implications for the pathogenesis of endometriosis. Hum Reprod. 2012;27:394–407.\nLiu L, Qi L, Knifley T, Piecoro DW, Rychahou P, Liu J, Mitov MI, Martin J, Wang C, Wu J, Weiss HL, Butterfield DA, Evers BM, O’Connor KL, Chen M. S100A4 alters metabolism and promotes invasion of lung cancer cells by up-regulating mitochondrial complex I protein NDUFS2. J Biol Chem. 2019;294:7516–27.\nBettum IJ, Gorad SS, Barkovskaya A, Pettersen S, Moestue SA, Vasiliauskaite K, Tenstad E, Øyjord T, Risa Ø, Nygaard V, Mælandsmo GM, Prasmickaite L. Metabolic reprogramming supports the invasive phenotype in malignant melanoma. Cancer Lett. 2015;366:71-83.\nMartínez-Zaguilán R, Seftor EA, Seftor RE, Chu YW, Gillies RJ, Hendrix MJ. Acidic pH enhances the invasive behavior of human melanoma cells. Clin Exp Metastasis. 1996;14:176-86.\nColegio OR, Chu NQ, Szabo AL, Chu T, Rhebergen AM, Jairam V, Cyrus N, Brokowski CE, Eisenbarth SC, Phillips GM, Cline GW, Phillips AJ, Medzhitov R. Functional polarization of tumour-associated macrophages by tumour-derived lactic acid. Nature. 2014;513:559–63.\nAcknowledgements\nWe would like to thank all patients who agreed to participate in this study and all colleagues who helped with this research.\nFunding\nThis work was supported by grants from the National Natural Science Foundation of China (no. 82001524 and no. 8257063225 to Hengwei Liu), the Natural Science Foundation of Hubei Province (no. 2020CFB310 to Hengwei Liu), the Traditional Chinese Medicine Hospital of Wuxue City scientific research project (no. 2023A005 to Yanyun Wu).\nAuthor information\nAuthors and Affiliations\nContributions\nX.P.W and Y.C. contributed equally to this work. X.P.W and Y.C. completed the molecular biology experiments and data collection, analysis and was the major contributor in writing the manuscript. X.P.W., Q.Y.W. and X.W.W. contributed to clinical sample collection. X.P.W, Y.C, Q.Y.W, H.W.L and Y.Y.W reviewed and polished the manuscript. H.W.L and Y.Y.W designed the study and to data acquisition. All authors reviewed and approved the final manuscript.\nCorresponding authors\nEthics declarations\nEthics Approval and Consent to Participate\nApproval by Ethics Committee: The Research Ethics Committee of Tongji Medical College, Huazhong University of Science (IORG 140 No: IORG0003571). Human rights statements and informed consent: All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1964 and its later amendments. Informed consent was obtained from all patients for inclusion in the study.\nCompeting Interests\nThe authors declare that they have no competing interests.\nAdditional information\nPublisher'S Note\nSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.\nSupplementary Information\nBelow is the link to the electronic supplementary material.\nRights and permissions\nSpringer 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.\nAbout this article\nCite this article\nWang, X., Chen, Y., Wu, Q. et al. Integrated Analysis of Lactate-Related Genes Identifies S100A4 as a Novel Marker Promoting the Migration and Invasion of Endometrial Stromal Cell in Endometriosis. Reprod. Sci. 32, 2558–2573 (2025). https://doi.org/10.1007/s43032-025-01914-7\nReceived:\nAccepted:\nPublished:\nVersion of record:\nIssue date:\nDOI: https://doi.org/10.1007/s43032-025-01914-7","source_license":"CC0","license_restricted":false}