Specific Aim (SA)1 will identify the critical bacterial species within the 35 isolate mixture that are responsible for biofilm formation and tumorigenesis. SA2 will test the hypothesis that Interleukin-17 (IL-17) and other immune parameters are critical mediators of biofilm formation and biofilm-mediated tumorigenesis utilizing a combination of single-cell RNA sequencing, flow cytometry, and qRT-PCR. SA3 will utilize advanced mass spectrometry methods to determine metabolites associated with biofilms in both patient and mouse tissues and the localization of these metabolites in select samples. The experiments in this proposal will provide valuable information regarding the host and microbial requirements for biofilm formation and biofilm-mediated tumorigenesis that may lead to potential therapeutic targets or biomarkers for biofilms.