<b>CHARACTERIZING THE ANTICANCER POTENTIAL OF VAGINAL MICROBES AND METABOLITES</b>

<p dir="ltr">The vaginal microbiome plays critical roles in gynecologic health, cancer risk, and immune regulation, yet systematic methods to characterize therapeutic potential of microbial metabolites remain limited. This dissertation presents the development and application of the Pharmacobiome, a novel systems biology framework that evaluates functional similarities between vaginal microbes, microbial metabolites, and known pharmaceutical compounds based on host transcriptomic signatures. Through two complementary studies, we demonstrate the Pharmacobiome's capacity to identify previously unrecognized therapeutic applications of vaginal microbiome-derived factors.</p><p dir="ltr">The first study applied the Pharmacobiome framework to characterize anti-cancer potential of vaginal microbes and metabolites by integrating multi-omics datasets from HIV-negative women with drug screening transcriptomics. Analysis identified over 40 vaginal metabolites demonstrating transcriptional parallels to 28 anti-cancer compounds. Metabolomic profiling of cultured Lactobacillus crispatus, Lactobacillus iners, and Gardnerella vaginalis revealed that L. crispatus uniquely produces taurine, which exhibited selective cytotoxicity against endometrial cancer cells (HEC1A) at physiologically relevant concentrations (1-130 μM) while sparing normal endometrial stromal cells. The second study utilized the Pharmacobiome framework to investigate immunomodulatory potential of vaginal metabolites on macrophage polarization. Spearman correlation analyses linking 51 microbial taxa and 99 metabolites to 23,482 epithelial genes, followed by comparison with macrophage polarization signatures, identified trimethylamine-N-oxide (TMAO), hydroxyproline, and hexose as key immunomodulators. Experimental validation using differentiated THP-1 macrophages demonstrated that 10 μM TMAO induced robust, dose-dependent secretion of proinflammatory cytokines including TNF-α, IL-1β, IL-6, MIP-1β, RANTES, IP-10, and IL-12p70. These findings establish the Pharmacobiome as a powerful platform for discovering microbiome-based therapeutics with applications in cancer treatment and immune modulation.</p>