{"paper_id":"f6073898-179a-4515-b81d-1a565ee81116","body_text":"Abstract\nEndometriosis (EMs) affects the physical and mental health of a wide range of women of childbearing age. Exploring specific diagnostic targets and screening potential therapeutic drugs will help early diagnosis and improve prognosis. The diagnostic and therapeutic potential of ETS homologous factor (EHF) and Gamma linolenic acid (GLA) remains to be explored. Based on RNA sequencing of GEO datasets and local clinical samples, LASSO and SVM-RFE machine learning algorithms were used to screen out the key gene (EHF). Explore the expression level of EHF and its effects on the malignant biological behavior and regulatory mechanism of endometriosis cells (12z). Based on the positive and negative correlation genes of EHF, cMap analysis and molecular docking were performed to screen out the potential therapeutic drug (GLA). Explore the therapeutic effects of GLA on endometriosis in vivo and in vitro. EHF was abnormally low in endometriosis tissues and cells. Overexpression of EHF could inhibit malignant phenotypes such as cell proliferation, migration, invasion, and angiogenesis, and inhibit the PI3K/AKT/mTOR axis, leading to cell cycle arrest and inhibiting epithelial-mesenchymal transition (EMT). GLA could play a potential therapeutic role in vivo and in vitro with low normal cell toxicity. EHF is a potential protective gene for endometriosis, and its overexpression could inhibit the malignant biological phenotype, cell cycle and EMT by inhibiting the PI3K/AKT/mTOR axis. GLA screened based on EHF could be used as a potential therapeutic drug to play a therapeutic role in vitro and in vivo. This study screened and validated specific diagnostic targets (EHF) and potential therapeutic drugs (GLA) for endometriosis based on clinical tissue sample tissue, cells, RNA sequencing, in vivo and in vitro experiments to assist in the early diagnosis and prognosis improvement of EMs.\nSimilar content being viewed by others\nData availability\nThe data can be obtained through the email under reasonable request: zld8399@163.com.\nAbbreviations\n- Ems:\n-\nEndometriosis\n- EHF:\n-\nETS homologous factor\n- GLA:\n-\nGamma linolenic acid\n- GEO:\n-\nGene Expression Omnibus\n- RNA-seq:\n-\nRNA sequencing\n- LASSO:\n-\nLeast Absolute Shrinkage and Selection Operator\n- SVM:\n-\nSupport Vector Machines\n- GO:\n-\nGene Ontology\n- KEGG:\n-\nKyoto Encyclopedia of Genes and Genomes\n- GSEA:\n-\nGene Set Enrichment Analysis\n- 12z:\n-\nHuman immortalized endometriosis cells\n- hEEC:\n-\nHuman immortalized endometrial cells\n- cMap:\n-\nConnectivity Map\n- EMT:\n-\nEpithelial-Mesenchymal Transition\nReferences\nAlbino D, Civenni G, Dallavalle C et al (2016) Activation of the Lin28/let-7 axis by loss of ESE3/EHF promotes a tumorigenic and stem-like phenotype in prostate cancer. Cancer Res 76:3629–3643. https://doi.org/10.1158/0008-5472.CAN-15-2665\nAlbino D, Longoni N, Curti L et al (2012) ESE3/EHF controls epithelial cell differentiation and its loss leads to prostate tumors with mesenchymal and stem-like features. Cancer Res 72:2889–2900. https://doi.org/10.1158/0008-5472.CAN-12-0212\nArmignacco R, Carlier N, Jouinot A et al (2024) Whole blood transcriptome signature predicts severe forms of COVID-19: results from the COVIDef cohort study. Funct Integr Genomics 24:107. https://doi.org/10.1007/s10142-024-01359-2\nCao D, Luo J, Zang W et al (2016) Gamma-linolenic acid suppresses NF-kappaBeta signaling via CD36 in the lipopolysaccharide-induced inflammatory response in primary goat mammary gland epithelial cells. Inflammation 39:1225–1237. https://doi.org/10.1007/s10753-016-0358-7\nExcess Polyunsaturated Fatty Acids Induce Death in Acidic Cancer Cells. (2021). Cancer Discov 11: 1872. https://doi.org/10.1158/2159-8290.CD-RW2021-090\nFearon KC, Falconer JS, Ross JA et al (1996) An open-label phase I/II dose escalation study of the treatment of pancreatic cancer using lithium gammalinolenate. Anticancer Res 16:867–874\nFossum SL, Mutolo MJ, Tugores A et al (2017) Ets homologous factor (EHF) has critical roles in epithelial dysfunction in airway disease. J Biol Chem 292:10938–10949. https://doi.org/10.1074/jbc.M117.775304\nFossum SL, Mutolo MJ, Yang R et al (2014) Ets homologous factor regulates pathways controlling response to injury in airway epithelial cells. Nucleic Acids Res 42:13588–13598. https://doi.org/10.1093/nar/gku1146\nHooper G (1940) The diagnosis and treatment of endometriosis. Can Med Assoc J 42:243–246\nHorne AW, Saunders PTK (2019) Snapshot: endometriosis. Cell 179(7):e1. https://doi.org/10.1016/j.cell.2019.11.033\nJiang WG, Bryce RP, Horrobin DF (1998) Essential fatty acids: molecular and cellular basis of their anti-cancer action and clinical implications. Crit Rev Oncol Hematol 27:179–209. https://doi.org/10.1016/s1040-8428(98)00003-1\nKajiyama H, Suzuki S, Yoshihara M et al (2019) Endometriosis and cancer. Free Radic Biol Med 133:186–192. https://doi.org/10.1016/j.freeradbiomed.2018.12.015\nKamble P, Nagar PR, Bhakhar KA et al (2024) Cancer pharmacoinformatics: databases and analytical tools. Funct Integr Genomics 24:166. https://doi.org/10.1007/s10142-024-01445-5\nKas K, Finger E, Grall F et al (2000) ESE-3, a novel member of an epithelium-specific ets transcription factor subfamily, demonstrates different target gene specificity from ESE-1. J Biol Chem 275:2986–2998. https://doi.org/10.1074/jbc.275.4.2986\nKavanagh K, Flynn DM, Jenkins KA et al (2013) Stearidonic and γ-linolenic acids in echium oil improves glucose disposal in insulin resistant monkeys. Prostaglandins Leukot Essent Fatty Acids 89:39–45. https://doi.org/10.1016/j.plefa.2013.04.003\nKenny FS, Pinder SE, Ellis IO et al (2000) Gamma linolenic acid with tamoxifen as primary therapy in breast cancer. Int J Cancer 85:643–648. https://doi.org/10.1002/(sici)1097-0215(20000301)85:5%3c643::aid-ijc8%3e3.0.co;2-z\nKleinbaum LA, Duggan C, Ferreira E et al (1999) Human chromosomal localization, tissue/tumor expression, and regulatory function of the ets family gene EHF. Biochem Biophys Res Commun 264:119–126. https://doi.org/10.1006/bbrc.1999.1493\nLee TC, Ivester P, Hester AG et al (2014) The impact of polyunsaturated fatty acid-based dietary supplements on disease biomarkers in a metabolic syndrome/diabetes population. Lipids Health Dis 13:196. https://doi.org/10.1186/1476-511X-13-196\nLim DV, Woo WH, Lim JX et al (2024) Targeting Mutant-p53 for Cancer Treatment: Are We There Yet? Curr Mol Pharmacol 17:e140923221042. https://doi.org/10.2174/1874467217666230914090621\nLiu J, Jiang W, Zhao K et al (2019) Tumoral EHF predicts the efficacy of anti-PD1 therapy in pancreatic ductal adenocarcinoma. J Exp Med 216:656–673. https://doi.org/10.1084/jem.20180749\nLuk IY, Jenkins LJ, Schoffer KL et al (2022) Epithelial de-differentiation triggered by co-ordinate epigenetic inactivation of the EHF and CDX1 transcription factors drives colorectal cancer progression. Cell Death Differ 29:2288–2302. https://doi.org/10.1038/s41418-022-01016-w\nMustonen, A. M., and P. Nieminen. (2023) Dihomo-gamma-Linolenic Acid (20:3n-6)-Metabolism, Derivatives, and Potential Significance in Chronic Inflammation. Int J Mol Sci 24. https://doi.org/10.3390/ijms24032116\nNightingale R, Reehorst CM, Vukelic N et al (2024) Ehf controls mammary alveolar lineage differentiation and is a putative suppressor of breast tumorigenesis. Dev Cell. https://doi.org/10.1016/j.devcel.2024.04.022\nRahmioglu N, Mortlock S, Ghiasi M et al (2023) The genetic basis of endometriosis and comorbidity with other pain and inflammatory conditions. Nat Genet 55:423–436. https://doi.org/10.1038/s41588-023-01323-z\nRautiainen S, Manson JE, Lichtenstein AH et al (2016) Dietary supplements and disease prevention - a global overview. Nat Rev Endocrinol 12:407–420. https://doi.org/10.1038/nrendo.2016.54\nRice TW, Wheeler AP, Thompson BT et al (2011) Enteral omega-3 fatty acid, gamma-linolenic acid, and antioxidant supplementation in acute lung injury. JAMA 306:1574–1581. https://doi.org/10.1001/jama.2011.1435\nSergeant S, Rahbar E, Chilton FH (2016) Gamma-linolenic acid, dihommo-gamma linolenic, eicosanoids and inflammatory processes. Eur J Pharmacol 785:77–86. https://doi.org/10.1016/j.ejphar.2016.04.020\nSriram K (2011) Confusion between gamma-linolenic acid and gamma-linoleic acid. Crit Care Med 39:2789. https://doi.org/10.1097/CCM.0b013e31822e56e0\nTang JY, Yang L, Wu QJ et al (2024) Circ-IP6K2 suppresses tumor progression by modulating the miR-1292-5p/CAMK2N1 signal in clear cell renal cell carcinoma. Funct Integr Genomics 24:122. https://doi.org/10.1007/s10142-024-01398-9\nVercellini P, Vigano P, Somigliana E et al (2014) Endometriosis: pathogenesis and treatment. Nat Rev Endocrinol 10:261–275. https://doi.org/10.1038/nrendo.2013.255\nWang N, Miao X, Lu W et al (2024a) RUNX3 exerts tumor-suppressive role through inhibiting EXOSC4 expression. Funct Integr Genomics 24:103. https://doi.org/10.1007/s10142-024-01363-6\nWang Z, Zhao Y, Zhang L (2024b) Emerging trends and hot topics in the application of multi-omics in drug discovery: a bibliometric and visualized study. Curr Pharm Anal 21:20–32. https://doi.org/10.1016/j.cpan.2024.12.001\nXie Y, Zhou T, Li X et al (2024) Targeting ESE3/EHF with nifurtimox inhibits CXCR2(+) neutrophil infiltration and overcomes pancreatic cancer resistance to chemotherapy and immunotherapy. Gastroenterology. https://doi.org/10.1053/j.gastro.2024.02.046\nZhao T, Jiang W, Wang X et al (2017) Ese3 inhibits pancreatic cancer metastasis by upregulating E-cadherin. Cancer Res 77:874–885. https://doi.org/10.1158/0008-5472.CAN-16-2170\nZhou T, Liu J, Xie Y et al (2022) ESE3/EHF, a promising target of rosiglitazone, suppresses pancreatic cancer stemness by downregulating CXCR4. Gut 71:357–371. https://doi.org/10.1136/gutjnl-2020-321952\nZhou X, Tan F, Zhang S et al (2023) Combining single-cell RNA sequencing data and transcriptomic data to unravel potential mechanisms and signature genes of the progression of idiopathic pulmonary fibrosis to lung adenocarcinoma and predict therapeutic agents. Funct Integr Genomics 23:346. https://doi.org/10.1007/s10142-023-01274-y\nZhu Y (2024) Plasma/serum proteomics based on mass spectrometry. Protein Pept Lett 31:192–208. https://doi.org/10.2174/0109298665286952240212053723\nZondervan KT, Becker CM, Koga K et al (2018) Endometriosis. Nat Rev Dis Primers 4:9. https://doi.org/10.1038/s41572-018-0008-5\nZondervan KT, Becker CM, Missmer SA (2020) Endometriosis. N Engl J Med 382:1244–1256. https://doi.org/10.1056/NEJMra1810764\nAcknowledgements\nThanks for the support of The Third Affiliated Hospital of Zhengzhou University.\nFunding\nThis work is supported by PhD research startup foundation of the Third Affiliated Hospital of Zhengzhou University (BS20230104). Henan Province Science and Technology Research Project (No. LHGJ20230372 and No. LHGJ20230051). National Natural Science Foundation of China (No. 82403992). National Natural Science Foundation of Henan (No. 242300421492).\nAuthor information\nAuthors and Affiliations\nContributions\nMengjun Zhang and Jialin Wang contributed equally to this work. Design of the study were performed by MZ, XZ and LZ. Implementation of the experiment were performed by MZ, HY and JW. Data analysis were performed by ZZ. The first draft of the manuscript was written and revised by MZ, XZ and LZ. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.\nCorresponding authors\nEthics declarations\nEthical approval and consent to participate\nThe Ethics Review Committee of the Third Affiliated Hospital of Zhengzhou University reviewed and approved this study (No.: 2023–098; Date: April 28, 2023). All patients read and signed the informed consent form.\nClinical trial number\nNot applicable.\nConsent for publication\nNot applicable.\nCompeting interests\nThe authors declare 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\nZhang, M., Wang, J., Yue, H. et al. ETS Homologous Factor (EHF) and Gamma Linolenic Acid (GLA): novel strategies for early diagnosis and treatment of endometriosis. Funct Integr Genomics 26, 14 (2026). https://doi.org/10.1007/s10142-025-01792-x\nReceived:\nRevised:\nAccepted:\nPublished:\nVersion of record:\nDOI: https://doi.org/10.1007/s10142-025-01792-x","source_license":"CC0","license_restricted":false}