Metabolomic and microbiome analysis of cervicovaginal mucus in in vitro fertilization–embryo transfer: Toward predicting pregnancy success

Purpose: In the context of in vitro fertilization–embryo transfer (IVF–ET), factors other than egg quality may be key determinants of treatment success, in particular, maternal factors related to uterine endometrial receptivity and unidentified factors. We therefore aimed to analyze the metabolome and microbiome in IVF–ET patients who did and did not achieve pregnancy. Methods: Cervicovaginal mucus was collected from patients undergoing IVF–ET. Metabolite analysis was conducted by liquid chromatography-mass spectrometry and the microbiota were determined by the polymerase chain reaction using universal 16S-rRNA gene bacterial primers by MiSeq sequencing. Patients were classified as pregnant (N = 10) or nonpregnant (N = 13). Metabolic pathways were examined by MetaboAnalyst. Results: Three metabolic pathways, including alanine-aspartate–glutamate metabolism, arginine biosynthesis, and cysteine-methionine metabolism, were commonly decreased at the time of embryo transfer irrespective pregnant outcomes. Notably, pyruvate was decreased in the pregnant group. Amino acid metabolites showed inverse correlations with the presence of anaerobic microbiota in the nonpregnant group. Conclusions: Metabolism decreased during embryo transplantation, with a notable decrease in pyruvate metabolism, particularly in patients who became pregnant. The behavior of metabolites in the pregnant and nonpregnant groups suggests that metabolome analysis in the cervicovaginal mucus may be a diagnostic marker for predicting pregnancy.