Metabolic changes in follicular fluids of patients treated with recombinant versus urinary human chorionic gonadotropin for triggering ovulation in assisted reproductive technologies: a metabolomics pilot study

Abstract Introduction The main goal of this retrospective cohort study is the assessment of the effects of administration of recombinant- hCG (r-hCG) versus urinary-hCG (u-hCG) on follicular fluid (FF) composition of women who underwent in vitro fertilization (IVF) treatments. Materials and methods We selected 70 patients with infertility attributable to tubal diseases, unexplained infertility, and male factor. Metabolomics analysis of their FFs was performed by 1H nuclear magnetic resonance (1H NMR) spectroscopy in combination with multivariate analysis to interpret the spectral data. Univariate statistical analysis was applied to investigate the possible correlations between clinical parameters and between clinical parameters and metabolites identified by NMR. Results According to the type of hCG used, significant differences were detected in FFs of women with male factor and unexplained infertility, both in qualitative and quantitative terms, for some metabolites as cholesterol, citrate, creatine, β-hydroxybutyrate, glycerol, lipids, amino acids (Glu, Gln, His, Val, Lys) and glucose. No significant difference was observed in women with tubal diseases. Besides, the number of MII oocytes in the u-hCG-treated groups correlates positively with glutamate in tubal disease and with glycerol in unexplained infertility. In the r-hCG-treated groups, the number of MII oocytes correlates positively with lipid in tubal disease, positively with citrate and negatively with glucose in male infertility. Conclusions Metabolite composition of FF changes according to different type of hCG treatment and this can be related to oocyte development and subsequent outcome. According to the data of this study, different types of hCG should be used in relation to the diagnosis of infertility to obtain better results in inducing oocyte maturation in women undergoing IVF

NMR metabolic profiling of follicular fluid for investigating the different causes of female infertility: a pilot study

Introduction Several metabolomics studies have correlated follicular fluid (FF) metabolite composition with oocyte competence to fertilization, embryo development and pregnancy but there is a scarcity of research examining the metabolic effects of various gynaecological diseases. Objectives In this study we aimed to analyze and correlate the metabolic profile of FF from women who were following in vitro fertilization (IVF) treatments with their different infertility pathologies. Methods We selected 53 women undergoing IVF who were affected by: tubal diseases, unexplained infertility, endometriosis, polycystic ovary syndrome (PCOS). FF of the study participants was collected at the time of oocytes retrieval. Metabolomic analysis of FF was performed by nuclear magnetic resonance (NMR) spectroscopy. Results FF presents some significant differences in various infertility pathologies. Although it was not possible to discriminate between FF of control participants and women with tubal diseases and unexplained infertility, comparison of FF metabolic profile from control women with patients with endometriosis and PCOS revealed significant differences in some metabolites that can be correlated to the causes of infertility. Conclusion NMR-based metabolic profiling may be successfully applied to find diagnostic biomarkers for PCOS and endometriosis and it might be also used to predict oocyte developmental potential and subsequent outcome

NMR metabolomics study of follicular fluid in women with cancer resorting to fertility preservation

Purpose The purpose of this study was to evaluate the possible application of metabolomics to identify follicular fluid changes in cancer patients undergoing fertility preservation. Although metabolomics have been applied already in cancer studies, this is the first application on follicular fluid of cancer patients. Methods We selected for the study ten patients with breast cancer and lymphoma who resorted to oocyte cryopreservation to preserve fertility and ten healthy women undergoing in vitro fertilization treatments. Follicular fluid was collected at the time of oocytes retrieval. Metabolomic analysis of follicular fluids was performed by 1H-nuclear magnetic resonance (NMR) spectroscopy in combination with multivariate analysis to interpret the spectral data. Univariate statistical analysis was applied to find correlations between patients’ features and metabolites identified by NMR. Results Partial least squares discriminant analysis allowed to discriminate samples from cancer patients and healthy controls. Univariate statistical analysis found significant correlations between patients’ features and metabolites identified by NMR. This finding allowed to identify biomarkers to differentiate both healthy controls from cancer patients and the two different classes of oncological patients. Conclusion The follicular fluids of cancer patients display significant metabolic alterations in comparison to healthy subjects. NMR-based metabolomics could be a valid prognostic tool for identifying and selecting the best cryopreserved oocytes and improving the outcome prediction in cancer women undergoing in vitro fertilization

METABOLOMICS STUDY OF FOLLICULAR FLUID IN WOMEN UNDERGOING IN VITRO FERTILIZATION (IVF)

Follicular fluid (FF) is the in-vivo oocyte environment that fills the antrum of a mature follicle, and contains important metabolites for oocyte growth and development. In the last years, FF has gained increased interest as it is a superfluous and easily available product aspirated together with oocyte during standard IVF procedures (1). Several investigations were conducted on FF to find biomarkers of oocyte quality by using targeted analyses where a selective class of molecules is profiled. The use of single biochemical markers should better be replaced with a metabolomic approach to assess both oocyte (1,2) and embryo quality (3), and to identify biomarkers for predicting IVF outcome (4). The most used techniques for metabolic profiling are mass spectrometry and NMR. NMR has many advantages because is capable of simultaneously detecting a wide variety of metabolites with accuracy and reproducibility without elaborate sample preparation (5). Only few studies correlated the FF biomarkers with infertility causes (6-8). We present some applications of NMR metabolomics in the study of FF in women who underwent IVF treatments. Significant correlations between patients' features and metabolites identified were observed both for cancer patients (9) and in women with different infertility pathologies (10). NMR-based metabolomics could be a valid prognostic tool for identifying and selecting the best cryopreserved oocytes and to identify metabolites correlated to the causes of infertility. References 1) Revelli et al Reprod Biol Endocrinol 2009 7:40 2) Piñero-Sagredo et al NMR Biomed 2010 23 485-95 3) McRae et al Int J Reprod Med 2013 2013:603167 4) Bracewell-Milnes et al Hum Reprod Update 2017 23 723-36 5) Markley et al Curr Opin Biotechnol 2017 43 34-40 6) Arya et al Med Hypotheses 2012 78 475-8 7) Zhang et al Oncotarget 2017 8 80472-80 8) Karaer et al Syst Biol Reprod Med 1-9 9) Castiglione Morelli et al JARG 2018 1-8 10) Castiglione Morelli et al Metabolomics 2019 15 1