AlkB Homolog 5 Regulates Hexokinase 2-Mediated Glycolysis and Participates in the Progression of Endometriosis

Endometriosis is a common gynecological endocrine disease with unclear pathogenesis. Evidence suggests enhanced aerobic glycolysis in ectopic endometrium of endometriosis. The role of N6-methyladenosine (m6A) modification in female reproductive diseases has been revealed in recent years, and it is involved in the regulation of glycolysis in a variety of diseases. Here, we investigated the regulatory effect of m6A modification on glycolysis and its role in endometriosis. RNA sequencing of ectopic endometrium of endometriosis and normal endometrium revealed that hexokinase 2 (HK2) a glycolysis-related gene, was significantly up-regulated in ectopic endometrium of endometriosis. Meanwhile, this result was supported by immunohistochemistry. Subsequently, we found that AlkB homolog 5 (ALKBH5) could upregulate HK2 in human endometrial stromal cells (THESCs). Up-regulation of ALKBH5 promoted glycolysis, invasion, and migration of THESCs, which could be alleviated by 2-Deoxy-d-glucose (2-DG). Furthermore, knockdown of HK2 in THESCs overexpressing ALKBH5 significantly attenuated the promoting effects of ALKBH5 on glycolysis, migration, and invasion of THESCs. Moreover, an ALKBH5 inhibitor, 5-Carboxy-8-hydroxyquinoline (IOX1) was found to inhibit the progression of endometriosis and glycolysis in a mouse model of endometriosis. In conclusion, ALKBH5 promoted glycolysis by up-regulating HK2 and contributed to the progression of endometriosis. ALKBH5 may be a new target for the treatment of endometriosis. Graphical Abstract ALKBH5 targeted HK2 to regulate glycolysis and promote migration and invasion of endometrial stromal cells. Disclosure The article's conclusions, implications, or opinions are the consensus of all the authors. Conflicts of Interest The authors declare no conflicts of interest. Data Availability Statement The data that support the findings of this study are available from the corresponding author on reasonable request. Supporting Information | Filename | Description | |---|---| | fsb270813-sup-0001-FigureS1.docxWord 2007 document , 832.8 KB | Figure S1. | | fsb270813-sup-0002-FigureS2.docxWord 2007 document , 387.8 KB | Figure S2. | | fsb270813-sup-0003-TextS1.pdfPDF document, 435 KB | Text S1. | | fsb270813-sup-0004-TextS2.docWord document, 94.5 KB | Text S2. | Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

- 1A. L. Zippl, E. Reiser, and B. Seeber, “Endometriosis and Mental Health Disorders: Identification and Treatment as Part of a Multimodal Approach,” Fertility and Sterility 121 (2024): 370–378. - 2C. M. Becker, A. Bokor, O. Heikinheimo, et al., “ESHRE guideline: endometriosis,” Human Reproduction Open 2022 (2022): hoac009. - 3L. Coxon, L. Demetriou, and K. Vincent, “Current Developments in Endometriosis-Associated Pain,” Cell Reports Medicine 5 (2024): 101769. - 4P. S. Thiel, O. Bougie, J. Pudwell, J. Shellenberger, M. P. Velez, and A. Murji, “Endometriosis and Mental Health: A Population-Based Cohort Study,” American Journal of Obstetrics and Gynecology 230 (2024): 649.e1–649.e19. - 5P. T. K. Saunders and A. W. Horne, “Endometriosis: Etiology, Pathobiology, and Therapeutic Prospects,” Cell 184 (2021): 2807–2824. - 6H. S. Taylor, A. M. Kotlyar, and V. A. Flores, “Endometriosis Is a Chronic Systemic Disease: Clinical Challenges and Novel Innovations,” Lancet 397 (2021): 839–852. - 7S. E. Bulun, “Endometriosis Caused by Retrograde Menstruation: Now Demonstrated by DNA Evidence,” Fertility and Sterility 118 (2022): 535–536. - 8H. Kobayashi, H. Shigetomi, and S. Imanaka, “Nonhormonal Therapy for Endometriosis Based on Energy Metabolism Regulation,” Reproduction and Fertility 2 (2021): C42–c57. - 9Y. Wang and G. J. Patti, “The Warburg Effect: A Signature of Mitochondrial Overload,” Trends in Cell Biology 33 (2023): 1014–1020. - 10M. Liao, D. Yao, L. Wu, et al., “Targeting the Warburg Effect: A Revisited Perspective From Molecular Mechanisms to Traditional and Innovative Therapeutic Strategies in Cancer,” Acta Pharmaceutica Sinica B 14 (2024): 953–1008. - 11E. P. Khashchenko, M. Y. Vysokikh, M. V. Marey, et al., “Altered Glycolysis, Mitochondrial Biogenesis, Autophagy and Apoptosis in Peritoneal Endometriosis in Adolescents,” International Journal of Molecular Sciences 25 (2024): 4238. - 12C. Lu, P. Qiao, R. Fu, et al., “Phosphorylation of PFKFB4 by PIM2 Promotes Anaerobic Glycolysis and Cell Proliferation in Endometriosis,” Cell Death & Disease 13 (2022): 790. - 13S. Wang, C. Jiang, K. Cao, R. Li, Z. Gao, and Y. Wang, “HK2 in Microglia and Macrophages Contribute to the Development of Neuropathic Pain,” Glia 72 (2024): 396–410. - 14K. A. Toniyan, A. A. Malkov, N. S. Biryukov, E. Y. Gorbacheva, V. V. Boyarintsev, and I. V. Ogneva, “The Cellular Respiration of Endometrial Biopsies From Patients With Various Forms of Endometriosis,” International Journal of Molecular Sciences 25 (2024): 3680. - 15S. Hou, S. Lei, H. Peng, et al., “Downregulating HK2 Inhibits Proliferation of Endometrial Stromal Cells Through a Noncanonical Pathway Involving Phosphorylation of Signal Transducer and Activator of Transcription 1 in Endometriosis,” Biology of Reproduction 107 (2022): 488–499. - 16E. Huang and L. Chen, “RNA N(6)-methyladenosine Modification in Female Reproductive Biology and Pathophysiology,” Cell Communication and Signaling 21 (2023): 53. - 17L. Jiang, M. Zhang, J. Wu, et al., “Exploring Diagnostic m6A Regulators in Endometriosis,” Aging (Albany NY) 12 (2020): 25916–25938. - 18S. Zhao, B. Zhang, H. Yuan, et al., “IGF2BP2 Promotes the Progression of Ovarian Endometriosis by Regulating m6A-Modified MEIS2 and GATA6,” International Journal of Biochemistry & Cell Biology 152 (2022): 106296. - 19X. Li, W. Xiong, X. Long, et al., “Inhibition of METTL3/m6A/miR126 Promotes the Migration and Invasion of Endometrial Stromal Cells in Endometriosis†,” Biology of Reproduction 105 (2021): 1221–1233. - 20J. Chen, Y. Fang, Y. Xu, and H. Sun, “Role of m6A Modification in Female Infertility and Reproductive System Diseases,” International Journal of Biological Sciences 18 (2022): 3592–3604. - 21S. W. Yue, H. L. Liu, H. F. Su, et al., “m6A-Regulated Tumor Glycolysis: New Advances in Epigenetics and Metabolism,” Molecular Cancer 22 (2023): 137. - 22Q. Wang, X. Guo, L. Li, et al., “N(6)-Methyladenosine METTL3 Promotes Cervical Cancer Tumorigenesis and Warburg Effect Through YTHDF1/HK2 Modification,” Cell Death & Disease 11 (2020): 911. - 23C. Shen, B. Xuan, T. Yan, et al., “m(6)A-Dependent Glycolysis Enhances Colorectal Cancer Progression,” Molecular Cancer 19 (2020): 72. - 24M. Ye, J. Chen, F. Lu, et al., “Down-Regulated FTO and ALKBH5 Co-Operatively Activates FOXO Signaling Through m6A Methylation Modification in HK2 mRNA Mediated by IGF2BP2 to Enhance Glycolysis in Colorectal Cancer,” Cell & Bioscience 13 (2023): 148. - 25K. Peng, R. P. Xia, F. Zhao, et al., “ALKBH5 Facilitates the Progression of Infantile Hemangioma by Increasing FOXF1 Expression in a m(6)A-YTHDF2 Dependent Manner to Activate HK-2 Signaling,” Molecular and Cellular Biochemistry 479 (2024): 3153–3166. - 26Y. Xu, H. Liu, W. Xiong, et al., “A Novel Mechanism Regulating Pyroptosis-Induced Fibrosis in Endometriosis via Lnc-MALAT1/miR-141-3p/NLRP3 Pathway†,” Biology of Reproduction 109 (2023): 156–171. - 27Z. Zhang, Y. Wang, L. Zeng, et al., “miR-218-5p in Endometrial Microenvironment Prevents the Migration of Ectopic Endometrial Stromal Cells by Inhibiting LASP1,” Reproductive Biology and Endocrinology 20 (2022): 64. - 28M. Wang, R. Fan, J. Jiang, et al., “PIM2 Promotes the Development of Ovarian Endometriosis by Enhancing Glycolysis and Fibrosis,” Reproductive Sciences 30 (2023): 2692–2702. - 29B. Xie, J. Lin, X. Chen, et al., “CircXRN2 Suppresses Tumor Progression Driven by Histone Lactylation Through Activating the Hippo Pathway in Human Bladder Cancer,” Molecular Cancer 22 (2023): 151. - 30J. Chen, C. Xu, K. Yang, et al., “Inhibition of ALKBH5 Attenuates I/R-Induced Renal Injury in Male Mice by Promoting Ccl28 m6A Modification and Increasing Treg Recruitment,” Nature Communications 14 (2023): 1161. - 31P. Cheng, H. Han, F. Chen, et al., “Amelioration of Acute Myocardial Infarction Injury Through Targeted Ferritin Nanocages Loaded With an ALKBH5 Inhibitor,” Acta Biomaterialia 140 (2022): 481–491. - 32S. Paul, S. Ghosh, and S. Kumar, “Tumor Glycolysis, an Essential Sweet Tooth of Tumor Cells,” Seminars in Cancer Biology 86 (2022): 1216–1230. - 33F. Cuffaro, E. Russo, and A. Amedei, “Endometriosis, Pain, and Related Psychological Disorders: Unveiling the Interplay Among the Microbiome, Inflammation, and Oxidative Stress as a Common Thread,” International Journal of Molecular Sciences 25 (2024): 6473. - 34S. Vannuccini, S. Clemenza, M. Rossi, and F. Petraglia, “Hormonal Treatments for Endometriosis: The Endocrine Background,” Reviews in Endocrine & Metabolic Disorders 23 (2022): 333–355. - 35W. Park, W. Lim, M. Kim, et al., “Female Reproductive Disease, Endometriosis: From Inflammation to Infertility,” Molecules and Cells 48 (2025): 100164. - 36J. Lu, X. Ling, L. Liu, et al., “Emerging Hallmarks of Endometriosis Metabolism: A Promising Target for the Treatment of Endometriosis,” Biochimica et Biophysica Acta, Molecular Cell Research 1870 (2023): 119381. - 37R. M. Marquardt, D. N. Tran, B. A. Lessey, M. S. Rahman, and J. W. Jeong, “Epigenetic Dysregulation in Endometriosis: Implications for Pathophysiology and Therapeutics,” Endocrine Reviews 44 (2023): 1074–1095. - 38S. Kasvandik, K. Samuel, M. Peters, et al., “Deep Quantitative Proteomics Reveals Extensive Metabolic Reprogramming and Cancer-Like Changes of Ectopic Endometriotic Stromal Cells,” Journal of Proteome Research 15 (2016): 572–584. - 39L. Alberghina, “The Warburg Effect Explained: Integration of Enhanced Glycolysis With Heterogeneous Mitochondria to Promote Cancer Cell Proliferation,” International Journal of Molecular Sciences 24 (2023): 15787. - 40Y. Gou, H. Wang, T. Wang, et al., “Ectopic Endometriotic Stromal Cells-Derived Lactate Induces M2 Macrophage Polarization via Mettl3/Trib1/ERK/STAT3 Signalling Pathway in Endometriosis,” Immunology 168 (2023): 389–402. - 41A. Schurr, “How the ‘Aerobic/Anaerobic Glycolysis’ Meme Formed a ‘Habit of Mind’ Which Impedes Progress in the Field of Brain Energy Metabolism,” International Journal of Molecular Sciences 25 (2024): 1433. - 42Q. Qiao, S. Hu, and X. Wang, “The Regulatory Roles and Clinical Significance of Glycolysis in Tumor,” Cancer Communications 44 (2024): 761–786. - 43J. Fang, S. Luo, and Z. Lu, “HK2: Gatekeeping Microglial Activity by Tuning Glucose Metabolism and Mitochondrial Functions,” Molecular Cell 83 (2023): 829–831. - 44H. Liu, J. Liang, X. Dai, et al., “Transcriptome-Wide N6-Methyladenosine (m6A) Methylation Profiling of Long Non-Coding RNAs in Ovarian Endometriosis,” Genomics 116 (2024): 110803. - 45F. Li, C. He, H. Yao, et al., “Glutamate From Nerve Cells Promotes Perineural Invasion in Pancreatic Cancer by Regulating Tumor Glycolysis Through HK2 mRNA-m6A Modification,” Pharmacological Research 187 (2023): 106555. - 46K. D. Meyer, Y. Saletore, P. Zumbo, O. Elemento, C. E. Mason, and S. R. Jaffrey, “Comprehensive Analysis of mRNA Methylation Reveals Enrichment in 3′ UTRs and Near Stop Codons,” Cell 149 (2012): 1635–1646. - 47J. Pi, J. Liu, H. Chang, et al., “Therapeutic Efficacy of ECs Foxp1 Targeting Hif1α-Hk2 Glycolysis Signal to Restrict Angiogenesis,” Redox Biology 75 (2024): 103281. - 48D. R. Morris, Y. Qu, A. Agrawal, R. P. Garofalo, and A. Casola, “HIF-1α Modulates Core Metabolism and Virus Replication in Primary Airway Epithelial Cells Infected With Respiratory Syncytial Virus,” Viruses 12 (2020): 1088. - 49L. Chen, X. Li, Y. Deng, et al., “The PI3K-Akt-mTOR Pathway Mediates Renal Pericyte-Myofibroblast Transition by Enhancing Glycolysis Through HKII,” Journal of Translational Medicine 21 (2023): 323. - 50C. Bao, S. Zhu, K. Song, and C. He, “HK2: A Potential Regulator of Osteoarthritis via Glycolytic and Non-Glycolytic Pathways,” Cell Communication and Signaling: CCS 20 (2022): 132. - 51X. Wen, J. Zhang, Z. Xu, et al., “Highly Expressed lncRNA H19 in Endometriosis Promotes Aerobic Glycolysis and Histone Lactylation,” Reproduction (Cambridge, England) 168 (2024): e240018. - 52Y. Sun, S. Zhang, X. Zhang, et al., “AURKA Enhances the Glycolysis and Development of Ovarian Endometriosis Through ERβ,” Endocrinology 165 (2024): bqae018. - 53X. Gao, W. Shao, J. Wang, et al., “Integrin β3 Enhances Glycolysis and Increases Lactate Production in Endometriosis,” Journal of Reproductive Immunology 165 (2024): 104312. - 54Y. Sun, Q. Wang, M. Wang, et al., “CHIP Induces Ubiquitination and Degradation of HMGB1 to Regulate Glycolysis in Ovarian Endometriosis,” Cellular and Molecular Life Sciences 80 (2022): 13. - 55M. A. Riaz, E. O. Mecha, C. O. A. Omwandho, F. Zeppernick, I. Meinhold-Heerlein, and L. Konrad, “The Different Gene Expression Profile in the Eutopic and Ectopic Endometrium Sheds New Light on the Endometrial Seed in Endometriosis,” Biomedicine 12 (2024): 1276. - 56J. Keckstein, E. Saridogan, U. A. Ulrich, et al., “The #Enzian Classification: A Comprehensive Non-Invasive and Surgical Description System for Endometriosis,” Acta Obstetricia et Gynecologica Scandinavica 100 (2021): 1165–1175. - 57M. Alifano, R. Trisolini, A. Cancellieri, and J. F. Regnard, “Thoracic Endometriosis: Current Knowledge,” Annals of Thoracic Surgery 81 (2006): 761–769. - 58K. A. Burns, A. M. Pearson, J. L. Slack, et al., “Endometriosis in the Mouse: Challenges and Progress Toward a ‘Best Fit’ Murine Model,” Frontiers in Physiology 12 (2021): 806574. - 59E. Greaves, H. O. D. Critchley, A. W. Horne, and P. T. K. Saunders, “Relevant Human Tissue Resources and Laboratory Models for Use in Endometriosis Research,” Acta Obstetricia et Gynecologica Scandinavica 96 (2017): 644–658. - 60Y. Lyu, Y. Wang, H. Ding, and P. Li, “Hypoxia-Induced m6A Demethylase ALKBH5 Promotes Ovarian Cancer Tumorigenicity by Decreasing Methylation of the lncRNA RMRP,” American Journal of Cancer Research 13 (2023): 4179–4191. - 61W. Dai, R. Guo, X. Na, et al., “Hypoxia and the Endometrium: An Indispensable Role for HIF-1α as Therapeutic Strategies,” Redox Biology 73 (2024): 103205. - 62W. N. Li, M. H. Wu, and S. J. Tsai, “Hypoxia and Reproductive Health: The Role of Hypoxia in the Development and Progression of Endometriosis,” Reproduction 161 (2021): F19–f31. - 63X. Lin, Y. Dai, W. Gu, et al., “The Involvement of RNA N6-Methyladenosine and Histone Methylation Modification in Decidualization and Endometriosis-Associated Infertility,” Clinical and Translational Medicine 14 (2024): e1564. - 64M. E. Barnard, L. V. Farland, B. Yan, et al., “Endometriosis Typology and Ovarian Cancer Risk,” JAMA 332 (2024): 482–489. - 65H. S. Kim, T. H. Kim, H. H. Chung, and Y. S. Song, “Risk and Prognosis of Ovarian Cancer in Women With Endometriosis: A Meta-Analysis,” British Journal of Cancer 110 (2014): 1878–1890. Citing Literature Article Metrics Total unique accesses to an article’s full text in HTML or PDF/ePDF format.More metric information Scite metrics Explore this article's citation statements on scite.ai Share QR Code Generating QR code QR code copied to clipboard! Something went wrong while generating your QR code. Please try again in a moment. If the issue persists, refresh the page or contact support. Export citation Unable to load citation data. Please try again in a moment. How to cite Elkins, L. J., & Spiegelman, M. (2021). pyUserCalc: A revised Jupyter notebook calculator for uranium-series disequilibria in basalts. Earth and Space Science, 8, e2020EA001619. https://doi.org/10.1029/2020EA001619 Download Citation If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click on download. This feature enables you to download the bibliographic information (also called citation data, header data, or metadata) for the articles on our site. Citation manager file format Use the dropdown list to choose how to format the bibliographic data you're harvesting. Several citation manager formats are available, including EndNote and BibTex. You can then copy the formatted citation (as displayed) or download it as file, to your device. If the RefWorks format is chosen, the 'Download' button will be replaced with an option to directly export to RefWorks