Abstract
BackgroundVaginal dysbiosis has emerged as a key risk factor in HPV acquisition, persistence, and potentially cervical carcinogenesis. However, the biological mechanisms driving persistence and carcinogenesis have not been elucidated. Hence, our objective was to perform metabolic profiling of the cervicovaginal microenvironment to identify interactions between virus, host and microbes in the context of genital inflammation, dysplasia, and cancer.MethodsIn a multicenter study, metabolic profiles of 78 premenopausal, non-pregnant women with low-grade (LSIL) and high-grade squamous intraepithelial lesions (HSIL), invasive cervical cancer (ICC), or healthy controls (HPV-positive and -negative Ctrl) were analyzed using gas chromatography-mass spectrometry. Metabolome and vaginal microbiome datasets were integrated using state-of-the-art bioinformatic tools (PICRUSt, AMON, and MIMOSA). Hierarchical clustering analysis (HCA) and principal component analysis (PCA) were employed to reveal the influence of genital inflammation, patient groups, and microbiota on metabolic profiles. Receiver Operating Characteristics (ROC) analysis was used to discriminate metabolites for each patient group. Statistical differences were tested using ANOVA or Mann-Whitney U test.ResultsMetabolomes of ICC patients (n=468 metabolites) formed a distinct cluster on PCA and HCA plots, due to enrichment of membrane lipids. Amino acid and nucleotide metabolites were depleted in HPV-positive Ctrl, LSIL and HSIL groups (P<0.05). Microbial communities were predicted to alter amino acid and nucleotide metabolisms. Eicosenoate, 3-hydroxybutyrate, and oleate/vaccenate (AUC > 0.9, P<0.01) discriminated ICC from healthy patients. Sphingolipids and plasmalogens positively correlated with genital inflammation (Spearman’s rho > 0.7). Anti-inflammatory nucleotides, adenosine and cytosine positively correlated with Lactobacillus abundance (Spearman’s rho>0.5) and negatively correlated with genital inflammation (Spearman’s rho<-0.3). HCA of metabolites demonstrated that metabolic profiles were driven by cancer, genital inflammation and Lactobacillus dominance.ConclusionThe complex virus-host-microbe interplay within the cervicovaginal microenvironment lead to unique metabolic fingerprints that could be exploited for future development of diagnostics, preventatives or treatments to positively impact women’s health outcomes.DisclosureNo significant relationships.