Metabolomic profiling in follicular fluid of patients with infertility-related deep endometriosis

Introduction Endometriosis is an estrogen-dependent gynecological disease that causes infertility, and potential metabolomic biomarkers related to ovarian endometriosis and poor outcomes after assisted reproductive treatments are still lacking.

The present study analyzed the metabolomic profiling of follicular fluid samples from 40 patients undergoing in vitro fertilization.

The follicular fluid samples were classified as controls (n = 22) and endometriosis patients (n = 18). The samples were submitted to Bligh and Dyer protocol followed by metabolomics analysis by ultra-performance liquid chromatography mass spectrometry. Clinical data was assessed by Students’ T-test and metabolomics data was analyzed by multivariate statistics by MetaboAnalyst 3.0 to obtain intrinsic characteristics that allowed for groups discrimination. The Receiver Operating Characteristic curve was carried out for the proposed biomarkers, aiming to determine their specificity and sensitivity, as a set and individually.

From the metabolomic analysis, 20 ion masses were selected as potential biomarkers from principal component analysis, which showed that all biomarkers were more abundant in the endometriosis group when compared to controls. Tentative attribution was performed by lipid maps database, demonstrating that these potential biomarkers correspond to fatty acids, carnitines, monoacylglycerols, lysophosphatidic acids, lysophosphatidylglycerols, diacylglycerols, lysophosphatidylcholines, phosphatidylserine, lysophosphatidylinositols and Phosphatidic Acid.

The use of mass spectrometry-based metabolomics allowed for the identification of effective biomarkers for ovarian endometriosis, which may contribute for a better comprehension of the disease and how it affects the ovary, as well as assisting in the development of accessory tools for endometriosis diagnosis and infertility management. Similar content being viewed by others

Adamson, G. D., & Pasta, D. J. (1994) Surgical treatment of endometriosis-associated infertility: Meta-analysis compared with survival analysis. American Journal of Obstetrics and Gynecology, 171(6), 1488–1504. Ahn, S., Monsanto, S., Miller, C., Singh, S., Thomas, R., & Tayade, C. (2015). Pathophysiology and immune dysfunction in endometriosis. BioMedicine Research International, 2015, 795976. Bligh, E. G., & Dyer, W. J. (1959). A rapid method of total lipid extraction and purification. Can J Biochem Physiol, 37, 911–917. Bras, M., Queenan, B., & Susin, S. A. (2005). Programmed cell death via mitochondria: Different modes of dying. Biochemistry (Mosc), 70, 231–239. Bulun, S. E. (2009) Endometriosis. The New England Journal of Medicine, 360(3), 268–279. Chen, S. U., Chou, C. H., Lee, H., Ho, C. H., Lin, C. W., & Yang, Y. S. (2008). Lysophosphatidic acid up-regulates expression of interleukin-8 and -6 in granulosa-lutein cells through its receptors and nuclear factor-kappaB dependent pathways: Implications for angiogenesis of corpus luteum and ovarian hyperstimulation syndrome. The Journal of Clinical Endocrinology and Metabolism, 93, 935–943. Cordeiro, F. B., Cataldi, T. R., Perkel, K. J., do Vale Teixeira da Costa, L., Rochetti, R. C., Stevanato, J., et al. (2015). Lipidomics analysis of follicular fluid by ESI-MS reveals potential biomarkers for ovarian endometriosis. Journal of Assisted Reproduction and Genetics, 32(12), 1817–1825. Dutta, M., Joshi, M., Srivastava, S., Lodh, I., Chakravarty, B., & Chaudhury, K. (2012). A metabonomics approach as a means for identification of potential biomarkers for early diagnosis of endometriosis. Molecular Biosystems, 8(12), 3281–3287. Fortune, J. E. (1994). Ovarian follicular growth and development in mammals. Biology of Reproduction, 50, 225–232. Ghazi, N., Arjmand, M., Akbari, Z., Mellati, A. O., Saheb-Kashaf, H., & Zamani, Z. (2016). (1)H NMR-based metabolomics approaches as non-invasive tools for diagnosis of endometriosis. International Journal of Reproductive BioMedicine (Yazd), 14(1), 1–8. Giorgi, V. S., Da Broi, M. G., Paz, C. C., Ferriani, R. A., & Navarro, P.A. (2016) N-acetyl-cysteine and l-carnitine prevent meiotic oocyte damage induced by follicular fluid from infertile women with mild endometriosis. Reproductive Science, 23(3), 342–351. González-Comadran, M., Schwarze, J. E., Zegers-Hochschild, F., Souza, M. D., Carreras, R., & Checa, M.Á. (2017). The impact of endometriosis on the outcome of assisted reproductive technology. Reproductive Biology and Endocrinology: RB&E, 15(1), 8. Greene, A. D., Lang, S. A., Kendziorski, J. A., Sroga-Rios, J. M., Herzog, T. J., & Burns, K. A. (2016). Endometriosis: where are we and where are we going? Reproduction (Cambridge, England), 152(3), 63–78. Hickey, M., Ballard, K., & Farquhar, C. (2014). Endometriosis. BMJ, 348, g1752. Huang, J. Y., & Rosenwaks, Z. (2012) In vitro fertilisation treatment and factors affecting success. Best Practice & Research. Clinical Obstetrics & Gynaecology, 26(6), 777–788. Jungheim, E. S., Louden, E. D., Chi, M. M., Frolova, A. I., Riley, J. K., & Moley, K. H. (2011). Preimplantation exposure of mouse embryos to palmitic acid results in fetal growth restriction followed by catch-up growth in the offspring. Biology of Reproduction, 85, 678–683. Kruger, T. F., Menkveld, R., Stander, F. S., Lombard, C. J., Van der Merwe, J. P., van Zyl, J. A., & Smith, K. (1986). Sperm morphologic features as a prognostic factor in in vitro fertilization. Fertility and Sterility, 46(6), 1118–1123. Kuehnbaum, N. L., & Britz-McKibbin, P. (2013). New advances in separation science for metabolomics: Resolving chemical diversity in a post-genomic era. Chemical Reviews, 113, 2437–2468. Kyvelidou, C., Sotiriou, D., Antonopoulou, T., Tsagkaraki, M., Tserevelakis, G. J., Filippidis, G., & Athanassakis, I. (2016) l-Carnitine affects preimplantation embryo development toward infertility in mice. Reproduction (Cambridge, England), 152(4), 283–291. Letsiou, S., Peterse, D. P., Fassbender, A., Hendriks, M. M., van den Broek, N. J., Berger, R., et al. (2017). Endometriosis is associated with aberrant metabolite profiles in plasma. Fertility and Sterility, 107(3), 699–706. Lin, X. N., Wei, M. L., Tong, X. M., Xu, W. H., Zhou, F., Huang, Q. X., et al. (2012). Outcome of in vitro fertilization in endometriosis-associated infertility: A 5-year database cohort study. Chinese Medical Journal, 125(15), 2688–2693. Mansour, G., Abdelrazik, H., Sharma, R. K., Radwan, E., Falcone, T., & Agarwal, A. (2009). l-Carnitine supplementation reduces oocyte cytoskeleton damage and embryo apoptosis induced by incubation in peritoneal fluid from patients with endometriosis. Fertility and Sterility, 91, 2079–2086. Mizuno, H., Ueda, K., Kobayashi, Y., Tsuyama, N., Todoroki, K., Min, J. Z., & Toyo’oka, T. (2017). The great importance of normalization of LC-MS data for highly-accurate non-targeted metabolomics. Biomedical Chromatography: BMC, 31(1), e3864. Murphy, A. A., Santanam, N., Morales, A. J., & Parthasarathy, S. (1998). Lysophosphatidyl choline, a chemotactic factor for monocytes/T-lymphocytes is elevated in endometriosis. The Journal of Clinical Endocrinology and Metabolism, 83(6), 2110–2113. Rogers, P. A., D’Hooghe, T. M., Fazleabas, A., Gargett, C. E., Giudice, L. C., Montgomery, G. W., et al. (2009). Priorities for endometriosis research: Recommendations from an international consensus workshop. Reproductive Science, 16, 335–346. Sampson, J. (1927). Peritoneal endometriosis due to the menstrual dissemination of endometrial tissue into the peritoneal cavity. American Journal of Obstetrics and Gynecology, 14, 422–469. Tan, T. Y., Lau, S. K., Loh, S. F., & Tan, H. H. (2014). Female ageing and reproductive outcome in assisted reproduction cycles. Singapore Medical Journal, 55(6), 305–309. Vassiliadis, S., & Athanassakis, I. (2011). A “conditionally essential” nutrient, l-carnitine, as a primary suspect in endometriosis. Fertility and Sterility, 95(8), 2759–2760. Vouk, K., Hevir, N., Ribić-Pucelj, M., Haarpaintner, G., Scherb, H., Osredkar, J., et al. (2012). Discovery of phosphatidylcholines and sphingomyelins as biomarkers for ovarian endometriosis. Human Reproduction (Oxford, England), 27(10), 2955–2965. Vouk, K., Ribič-Pucelj, M., Adamski, J., & Rižner, T. L. (2016). Altered levels of acylcarnitines, phosphatidylcholines, and sphingomyelins in peritoneal fluid from ovarian endometriosis patients. The Journal of Steroid Biochemistry and Molecular Biology, 159, 60–69. Wang, J., Shen, X. X., Huang, X. H., & Zhao, Z. M. (2012). Follicular fluid levels of prostaglandin E2 and the effect of prostaglandin E2 on steroidogenesis in granulosa-lutein cells in women with moderate and severe endometriosis undergoing in vitro fertilization and embryo transfer. Chinese Medical Journal, 125, 3985–3990. Wang, Y., Liu, S., Hu, Y., Li, P., & Wan, J. (2015). Current state of the art of mass spectrometry-based metabolomics studies: A review focusing on wide coverage, high throughput and easy identification. RSC Advanced, 5, 78728–78737. Wei, Q., St Clair, J. B., Fu, T., Stratton, P., & Nieman, L. K. (2009). Reduced expression of biomarkers associated with the implantation window in women with endometriosis. Fertility and Sterility, 91(5), 1686–1691. Xia, J., & Wishart, D. S. (2016). Using metaboanalyst 3.0 for comprehensive metabolomics data analysis. Current Protocols in Bioinformatics, 55, 14.10.1–14.10.91. Ye, X., & Chun, J. (2010). Lysophosphatidic acid (LPA) signaling in vertebrate reproduction. Trends in Endocrinology and Metabolism: TEM, 21(1), 17–24. Funding The author Fernanda Bertuccez Cordeiro received scholarship from the National Counsel of Technological and Scientific Development—Brazil, during the development of this work. Author information Authors and Affiliations Corresponding author Ethics declarations Conflict of interest The authors declare that they have no conflict of interest. Ethical approval The present study received approval by Ethics in Research Committee of São Paulo Federal University and written informed consent was obtained from the participants. The procedures performed in this study for the patients’ treatment were independent of the study and the procedures performed for the study presented no influence on their treatment. The authors state that all procedures were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Electronic supplementary material Below is the link to the electronic supplementary material. Rights and permissions About this article Cite this article Cordeiro, F.B., Cataldi, T.R., do Vale Teixeira da Costa, L. et al. Metabolomic profiling in follicular fluid of patients with infertility-related deep endometriosis. Metabolomics 13, 120 (2017). https://doi.org/10.1007/s11306-017-1262-3 Received: Accepted: Published: DOI: https://doi.org/10.1007/s11306-017-1262-3