Ovarian Endometrioma Disrupts Oocyte-Cumulus Communication and Mitochondrial Function, With Melatonin Mitigating the Effects

Abstract

Ovarian endometrioma (OEM), a particularly severe form of endometriosis, is an oestrogen-dependent condition often associated with pain and infertility. The mechanisms by which OEM impairs fertility, particularly through its direct impact on oocyte-cumulus cell (CC) communication and related pathways, remain poorly understood. This study investigates the impact of OEM on oocyte-CC communication and explores melatonin's therapeutic potential. We used a mouse model of OEM and employed ovarian transcriptome and gene set enrichment analyses to identify disrupted gene pathways, alongside phalloidin staining for cytoskeletal analysis, gap junction coupling analysis for intercellular communication, and mitochondrial function assessments for cellular metabolism. Our results showed that OEM significantly impairs steroidogenesis and cumulus cell function, leading to increased apoptosis, disrupted transzonal projections (TZPs), and impaired antioxidant transfer to oocytes. This culminates in oxidative stress, mitochondrial dysfunction, and compromised ATP production. OEM oocytes also exhibited severe abnormalities, including DNA damage, maturation defects, spindle assembly disruptions, and increased aneuploidy. This study identifies disrupted TZPs as a key pathological feature in OEM and highlights melatonin's potential to restore intercellular communication, mitigate oxidative damage, and improve reproductive outcomes.

Keywords

melatonin / oocytes quality / ovarian endometriosis / ovarian infertility / reactive oxygen species / Transzonal projections

References

[1] P. Vercellini, P. Vigano, E. Somigliana, and L. Fedele, “Endometriosis: Pathogenesis and Treatment,” Nature Reviews. Endocrinology 10, no. 5 (2014): 261-275.

[2] R. Ruderman and M. E. Pavone, “Ovarian Cancer in Endometriosis: An Update on the Clinical and Molecular Aspects,” Minerva Ginecologica 69, no. 3 (2017): 286-294.

[3] S. Matsuzaki and B. Schubert, “Oxidative Stress Status in Normal Ovarian Cortex Surrounding Ovarian Endometriosis,” Fertility and Sterility 93, no. 7 (2010): 2431-2432.

[4] D. B. Sreerangaraja Urs, W. H. Wu, K. Komrskova, et al., “Mitochondrial Function in Modulating Human Granulosa Cell Steroidogenesis and Female Fertility,” International Journal of Molecular Sciences 21, no. 10 (2020): 3592.

[5] S. Gupta, A. Agarwal, R. Agarwal, and J. R. Loret de Mola, “Impact of Ovarian Endometrioma on Assisted Reproduction Outcomes,” Reproductive Biomedicine Online 13, no. 3 (2006): 349-360.

[6] S. K. Jana, K. NB, R. Chattopadhyay, B. Chakravarty, and K. Chaudhury, “Upper Control Limit of Reactive Oxygen Species in Follicular Fluid Beyond Which Viable Embryo Formation Is Not Favorable,” Reproductive Toxicology 29, no. 4 (2010): 447-451.

[7] G. Ragni, E. Somigliana, F. Benedetti, et al., “Damage to Ovarian Reserve Associated With Laparoscopic Excision of Endometriomas: A Quantitative Rather Than a Qualitative Injury,” American Journal of Obstetrics and Gynecology 193, no. 6 (2005): 1908-1914.

[8] D. Navot, P. A. Bergh, M. A. Williams, et al., “Poor Oocyte Quality Rather Than Implantation Failure as a Cause of Age-Related Decline in Female Fertility,” Lancet 337, no. 8754 (1991): 1375-1377.

[9] M. A. Edson, A. K. Nagaraja, and M. M. Matzuk, “The Mammalian Ovary From Genesis to Revelation,” Endocrine Reviews 30, no. 6 (2009): 624-712.

[10] J. Rojas, M. Chavez-Castillo, L. C. Olivar, et al., “Physiologic Course of Female Reproductive Function: A Molecular Look Into the Prologue of Life,” Journal of Pregnancy 2015 (2015): 715735.

[11] C. E. Plancha, A. Sanfins, P. Rodrigues, and D. Albertini, “Cell Polarity During Folliculogenesis and Oogenesis,” Reproductive Biomedicine Online 10, no. 4 (2005): 478-484.

[12] X. Lin, Y. Dai, X. Tong, et al., “Excessive Oxidative Stress in Cumulus Granulosa Cells Induced Cell Senescence Contributes to Endometriosis-Associated Infertility,” Redox Biology 30 (2020): 101431.

[13] D. A. Dumesic, D. R. Meldrum, M. G. Katz-Jaffe, R. L. Krisher, and W. B. Schoolcraft, “Oocyte Environment: Follicular Fluid and Cumulus Cells Are Critical for Oocyte Health,” Fertility and Sterility 103, no. 2 (2015): 303-316.

[14] D. F. Albertini, C. M. Combelles, E. Benecchi, and M. J. Carabatsos, “Cellular Basis for Paracrine Regulation of Ovarian Follicle Development,” Reproduction 121, no. 5 (2001): 647-653.

[15] G. Mansour, R. K. Sharma, A. Agarwal, and T. Falcone, “Endometriosis-Induced Alterations in Mouse Metaphase II Oocyte Microtubules and Chromosomal Alignment: A Possible Cause of Infertility,” Fertility and Sterility 94, no. 5 (2010): 1894-1899.

[16] H. Ferrero, A. Corachan, A. Aguilar, et al., “Single-Cell RNA Sequencing of Oocytes From Ovarian Endometriosis Patients Reveals a Differential Transcriptomic Profile Associated With Lower Quality,” Human Reproduction 34, no. 7 (2019): 1302-1312.

[17] M. Hamdan, K. T. Jones, Y. Cheong, and S. I. Lane, “The Sensitivity of the DNA Damage Checkpoint Prevents Oocyte Maturation in Endometriosis,” Scientific Reports 6 (2016): 36994.

[18] Z. Ren, J. Huang, C. Zhou, et al., “Transferrin and Antioxidants Partly Prevented Mouse Oocyte Oxidative Damage Induced by Exposure of Cumulus-Oocyte Complexes to Endometrioma Fluid,” Journal of Ovarian Research 13, no. 1 (2020): 139.

[19] H. Kobayashi, Y. Yamashita, A. Iwase, et al., “The Ferroimmunomodulatory Role of Ectopic Endometriotic Stromal Cells in Ovarian Endometriosis,” Fertility and Sterility 98, no. 2 (2012): 415-422 e411-412.

[20] O. Koc, B. Gunduz, A. Topcuoglu, G. Bugdayci, F. Yilmaz, and B. Duran, “Effects of Pinealectomy and Melatonin Supplementation on Endometrial Explants in a Rat Model,” European Journal of Obstetrics, Gynecology, and Reproductive Biology 153, no. 1 (2010): 72-76.

[21] Y. Li, S. W. Hung, R. Zhang, et al., “Melatonin in Endometriosis: Mechanistic Understanding and Clinical Insight,” Nutrients 14, no. 19 (2022): 4087.

[22] M. Guney, B. Oral, N. Karahan, and T. Mungan, “Regression of Endometrial Explants in a Rat Model of Endometriosis Treated With Melatonin,” Fertility and Sterility 89, no. 4 (2008): 934-942.

[23] B. Yilmaz, S. Kilic, O. Aksakal, et al., “Melatonin Causes Regression of Endometriotic Implants in Rats by Modulating Angiogenesis, Tissue Levels of Antioxidants and Matrix Metalloproteinases,” Archives of Gynecology and Obstetrics 292, no. 1 (2015): 209-216.

[24] J. Espino, M. Macedo, G. Lozano, et al., “Impact of Melatonin Supplementation in Women With Unexplained Infertility Undergoing Fertility Treatment,” Antioxidants (Basel) 8, no. 9 (2019): 338.

[25] A. Schwertner, C. C. Conceicao Dos Santos, G. D. Costa, et al., “Efficacy of Melatonin in the Treatment of Endometriosis: A Phase II, Randomized, Double-Blind, Placebo-Controlled Trial,” Pain 154, no. 6 (2013): 874-881.

[26] S. Hayashi, T. Nakamura, Y. Motooka, et al., “Novel Ovarian Endometriosis Model Causes Infertility via Iron-Mediated Oxidative Stress in Mice,” Redox Biology 37 (2020): 101726.

[27] A. Takeuchi, K. Koga, E. Satake, et al., “Endometriosis Triggers Excessive Activation of Primordial Follicles via PI3K-PTEN-Akt-Foxo3 Pathway,” Journal of Clinical Endocrinology and Metabolism 104, no. 11 (2019): 5547-5554.

[28] T. M. Leung and N. Nieto, “CYP2E1 and Oxidant Stress in Alcoholic and Non-Alcoholic Fatty Liver Disease,” Journal of Hepatology 58, no. 2 (2013): 395-398.

[29] C. Chen, S. Wang, L. Yu, et al., “H(2)O(2)-Mediated Autophagy During Ethanol Metabolism,” Redox Biology 46 (2021): 102081.

[30] H. Wu, Q. Liu, N. Yang, and S. Xu, “Polystyrene-Microplastics and DEHP Co-Exposure Induced DNA Damage, Cell Cycle Arrest and Necroptosis of Ovarian Granulosa Cells in Mice by Promoting ROS Production,” Science of the Total Environment 871 (2023): 161962.

[31] D. Richani, K. R. Dunning, J. G. Thompson, and R. B. Gilchrist, “Metabolic Co-Dependence of the Oocyte and Cumulus Cells: Essential Role in Determining Oocyte Developmental Competence,” Human Reproduction Update 27, no. 1 (2021): 27-47.

[32] M. Kitajima, M. M. Dolmans, O. Donnez, H. Masuzaki, M. Soares, and J. Donnez, “Enhanced Follicular Recruitment and Atresia in Cortex Derived From Ovaries With Endometriomas,” Fertility and Sterility 101, no. 4 (2014): 1031-1037.

[33] Y. Miao, Z. Cui, Q. Gao, R. Rui, and B. Xiong, “Nicotinamide Mononucleotide Supplementation Reverses the Declining Quality of Maternally Aged Oocytes,” Cell Reports 32, no. 5 (2020): 107987.

[34] H. Zhang, C. Li, D. Wen, et al., “Melatonin Improves the Quality of Maternally Aged Oocytes by Maintaining Intercellular Communication and Antioxidant Metabolite Supply,” Redox Biology 49 (2022): 102215.

[35] A. Bellon, L. Ortiz-Lopez, G. Ramirez-Rodriguez, F. Anton-Tay, and G. Benitez-King, “Melatonin Induces Neuritogenesis at Early Stages in N1E-115 Cells Through Actin Rearrangements via Activation of Protein Kinase C and Rho-Associated Kinase,” Journal of Pineal Research 42, no. 3 (2007): 214-221.

[36] P. T. Brower and R. M. Schultz, “Intercellular Communication Between Granulosa Cells and Mouse Oocytes: Existence and Possible Nutritional Role During Oocyte Growth,” Developmental Biology 90, no. 1 (1982): 144-153.

[37] V. Baena and M. Terasaki, “Three-Dimensional Organization of Transzonal Projections and Other Cytoplasmic Extensions in the Mouse Ovarian Follicle,” Scientific Reports 9, no. 1 (2019): 1262.

[38] E. Anderson and D. F. Albertini, “Gap Junctions Between the Oocyte and Companion Follicle Cells in the Mammalian Ovary,” Journal of Cell Biology 71, no. 2 (1976): 680-686.

[39] Y. Zhang, Y. Wang, X. Feng, et al., “Oocyte-Derived Microvilli Control Female Fertility by Optimizing Ovarian Follicle Selection in Mice,” Nature Communications 12, no. 1 (2021): 2523.

[40] A. D. Macaulay, I. Gilbert, J. Caballero, et al., “The Gametic Synapse: RNA Transfer to the Bovine Oocyte,” Biology of Reproduction 91, no. 4 (2014): 90.

[41] A. D. Macaulay, I. Gilbert, S. Scantland, et al., “Cumulus Cell Transcripts Transit to the Bovine Oocyte in Preparation for Maturation,” Biology of Reproduction 94, no. 1 (2016): 16.

[42] E. Winterhager and G. M. Kidder, “Gap Junction Connexins in Female Reproductive Organs: Implications for Women's Reproductive Health,” Human Reproduction Update 21, no. 3 (2015): 340-352.

[43] E. Hosseini, F. Mehraein, M. Shahhoseini, et al., “Epigenetic Alterations of CYP19A1 Gene in Cumulus Cells and Its Relevance to Infertility in Endometriosis,” Journal of Assisted Reproduction and Genetics 33, no. 8 (2016): 1105-1113.

[44] R. K. Pandya, S. R. Salian, V. R. Lakshmi, et al., “Global DNA Methylation Level in Spermatozoa Is Not Associated With ICSI Fertilization Outcome and Embryo Quality in Donor Oocyte Programme,” Human Fertility (Cambridge, England) 27, no. 1 (2024): 2421191.

[45] M. Samiec and M. Trzcinska, “From Genome to Epigenome: Who Is a Predominant Player in the Molecular Hallmarks Determining Epigenetic Mechanisms Underlying Ontogenesis?,” Reproductive Biology 24, no. 4 (2024): 100965.

[46] R. Xu, Q. Zhu, Y. Zhao, et al., “Unreprogrammed H3K9me3 Prevents Minor Zygotic Genome Activation and Lineage Commitment in SCNT Embryos,” Nature Communications 14, no. 1 (2023): 4807.

[47] Z. Ni, Y. Li, D. Song, et al., “Iron-Overloaded Follicular Fluid Increases the Risk of Endometriosis-Related Infertility by Triggering Granulosa Cell Ferroptosis and Oocyte Dysmaturity,” Cell Death & Disease 13, no. 7 (2022): 579.

[48] Y. Wu, R. Yang, J. Lan, et al., “Iron Overload Modulates Follicular Microenvironment via ROS/HIF-1alpha/FSHR Signaling,” Free Radical Biology & Medicine 196 (2023): 37-52.

[49] S. L. Barrett and D. F. Albertini, “Cumulus Cell Contact During Oocyte Maturation in Mice Regulates Meiotic Spindle Positioning and Enhances Developmental Competence,” Journal of Assisted Reproduction and Genetics 27, no. 1 (2010): 29-39.

[50] S. El-Hayek and H. J. Clarke, “Follicle-Stimulating Hormone Increases Gap Junctional Communication Between Somatic and Germ-Line Follicular Compartments During Murine Oogenesis,” Biology of Reproduction 93, no. 2 (2015): 47.

[51] S. Brunet and B. Maro, “Germinal Vesicle Position and Meiotic Maturation in Mouse Oocyte,” Reproduction 133, no. 6 (2007): 1069-1072.

[52] Y. Q. Su, K. Sugiura, and J. J. Eppig, “Mouse Oocyte Control of Granulosa Cell Development and Function: Paracrine Regulation of Cumulus Cell Metabolism,” Seminars in Reproductive Medicine 27, no. 1 (2009): 32-42.

[53] H. L. Xie, Y. B. Wang, G. Z. Jiao, et al., “Effects of Glucose Metabolism During In Vitro Maturation on Cytoplasmic Maturation of Mouse Oocytes,” Scientific Reports 6 (2016): 20764.

[54] A. Karaer, G. Tuncay, A. Mumcu, and B. Dogan, “Metabolomics Analysis of Follicular Fluid in Women With Ovarian Endometriosis Undergoing In Vitro Fertilization,” Systems Biology in Reproductive Medicine 65, no. 1 (2019): 39-47.

[55] J. M. Jimenez-Lopez and A. I. Cederbaum, “CYP2E1-Dependent Oxidative Stress and Toxicity: Role in Ethanol-Induced Liver Injury,” Expert Opinion on Drug Metabolism & Toxicology 1, no. 4 (2005): 671-685.

[56] J. Bai and A. I. Cederbaum, “Overexpression of CYP2E1 in Mitochondria Sensitizes HepG2 Cells to the Toxicity Caused by Depletion of Glutathione,” Journal of Biological Chemistry 281, no. 8 (2006): 5128-5136.

[57] H. Wang, C. Zhong, R. Yang, et al., “Hfm1 Participates in Golgi-Associated Spindle Assembly and Division in Mouse Oocyte Meiosis,” Cell Death & Disease 11, no. 6 (2020): 490.

[58] F. K. Boynukalin, M. Serdarogullari, M. Gultomruk, O. Coban, N. Findikli, and M. Bahceci, “The Impact of Endometriosis on Early Embryo Morphokinetics: A Case-Control Study,” Systems Biology in Reproductive Medicine 65, no. 3 (2019): 250-257.

[59] P. Stein and R. M. Schultz, “ICSI in the Mouse,” Methods in Enzymology 476 (2010): 251-262.

[60] S. Ferrero, G. Evangelisti, and F. Barra, “Current and Emerging Treatment Options for Endometriosis,” Expert Opinion on Pharmacotherapy 19, no. 10 (2018): 1109-1125.

[61] Y. Li, H. Liu, K. Wu, et al., “Melatonin Promotes Human Oocyte Maturation and Early Embryo Development by Enhancing Clathrin-Mediated Endocytosis,” Journal of Pineal Research 67, no. 3 (2019): e12601.

[62] K. H. Tsui, C. J. Li, and L. T. Lin, “Melatonin Supplementation Attenuates Cuproptosis and Ferroptosis in Aging Cumulus and Granulosa Cells: Potential for Improving IVF Outcomes in Advanced Maternal Age,” Reproductive Biology and Endocrinology 22, no. 1 (2024): 138.

[63] G. J. Lee, F. Porreca, and E. Navratilova, “Prolactin and Pain of Endometriosis,” Pharmacology & Therapeutics 247 (2023): 108435.

[64] J. A. Garcia-Velasco, N. G. Mahutte, J. Corona, et al., “Removal of Endometriomas Before In Vitro Fertilization Does Not Improve Fertility Outcomes: A Matched, Case-Control Study,” Fertility and Sterility 81, no. 5 (2004): 1194-1197.

[65] Y. Dai, X. Lin, W. Xu, et al., “MiR-210-3p Protects Endometriotic Cells From Oxidative Stress-Induced Cell Cycle Arrest by Targeting BARD1,” Cell Death & Disease 10, no. 2 (2019): 144.

[66] T. Motohara, S. Masuko, T. Ishimoto, et al., “Transient Depletion of p53 Followed by Transduction of c-Myc and K-Ras Converts Ovarian Stem-Like Cells Into Tumor-Initiating Cells,” Carcinogenesis 32, no. 11 (2011): 1597-1606.

[67] A. Freis, J. E. Dietrich, M. Binder, V. Holschbach, T. Strowitzki, and A. Germeyer, “Relative Morphokinetics Assessed by Time-Lapse Imaging Are Altered in Embryos From Patients With Endometriosis,” Reproductive Sciences 25, no. 8 (2018): 1279-1285.

[68] N. Garrido, J. Navarro, J. Remohi, C. Simon, and A. Pellicer, “Follicular Hormonal Environment and Embryo Quality in Women With Endometriosis,” Human Reproduction Update 6, no. 1 (2000): 67-74.

[69] J. S. Younis, “Is Oocyte Quality Impaired in Cases With Ovarian Endometriosis? A Second Look Into the Clinical Setting,” Front Endocrinol (Lausanne) 13 (2022): 921032.

[70] H. Wang, Z. Huang, X. Shen, et al., “Rejuvenation of Aged Oocyte Through Exposure to Young Follicular Microenvironment,” Nature Aging 4, no. 9 (2024): 1194-1210.

[71] Y. Yang, H. Zhao, M. Wu, et al., “3D-Printed Scaffolds With Controlled-Releasing Compounds for Ectopic Activation of Dormant Ovarian Follicles,” International Journal of Bioprinting (2024): 4941.

[72] M. Samiec, “Molecular Mechanism and Application of Somatic Cell Cloning in Mammals-Past, Present and Future,” International Journal of Molecular Sciences 23, no. 22 (2022): 13786.

[73] Y. G. Chung, S. Matoba, Y. Liu, et al., “Histone Demethylase Expression Enhances Human Somatic Cell Nuclear Transfer Efficiency and Promotes Derivation of Pluripotent Stem Cells,” Cell Stem Cell 17, no. 6 (2015): 758-766.

[74] M. Samiec and M. Skrzyszowska, “Biological Transcomplementary Activation as a Novel and Effective Strategy Applied to the Generation of Porcine Somatic Cell Cloned Embryos,” Reproductive Biology 14, no. 2 (2014): 128-139.

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2025 The Author(s). Cell Proliferation published by Beijing Institute for Stem Cell and Regenerative Medicine and John Wiley & Sons Ltd.