You have accessJournal of UrologyCME1 Apr 2023MP14-11 SINGLE CELL-BASED IMMUNE PROFILING OF THE TUMOR AND ITS IMMUNE MICROENVIRONMENT REVEALED DIFFERENCES BETWEEN NMIBC AND MIBC: IMPLICATION FOR IMMUNO-THERAPY Massimo Lazzeri, NicolòMaria Buffi, Giovanni Lughezzani, Paolo Casale, Roberto Hurle, Alberto Saita, Vittorio Fasulo, Marco Paciotti, Marta Pandini, Roberta Carriero, Mattia Carvetta, Marta Iovino, Piergiuseppe Colombo, Grazia Elefante, Gianluca Basso, Sergio Marchini, Paolo Kunderfranco, and Diletta Di Mitri Massimo LazzeriMassimo Lazzeri More articles by this author , NicolòMaria BuffiNicolòMaria Buffi More articles by this author , Giovanni LughezzaniGiovanni Lughezzani More articles by this author , Paolo CasalePaolo Casale More articles by this author , Roberto HurleRoberto Hurle More articles by this author , Alberto SaitaAlberto Saita More articles by this author , Vittorio FasuloVittorio Fasulo More articles by this author , Marco PaciottiMarco Paciotti More articles by this author , Marta PandiniMarta Pandini More articles by this author , Roberta CarrieroRoberta Carriero More articles by this author , Mattia CarvettaMattia Carvetta More articles by this author , Marta IovinoMarta Iovino More articles by this author , Piergiuseppe ColomboPiergiuseppe Colombo More articles by this author , Grazia ElefanteGrazia Elefante More articles by this author , Gianluca BassoGianluca Basso More articles by this author , Sergio MarchiniSergio Marchini More articles by this author , Paolo KunderfrancoPaolo Kunderfranco More articles by this author , and Diletta Di MitriDiletta Di Mitri More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003234.11AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Novel predictive tools to guide treatment decision are required and novel therapies are needed. Immunotherapies are under investigation and hold strong promise, however efficacy is still limited. On this line, a comprehensive analysis of the immune infiltrate in BC would reveal mechanisms of resistance and unveil novel targets. METHODS: Here we investigated the profile of the immune microenvironment in NMBC and MIBC patients by high dimensional single cell analysis, including scRNAseq and Imaging Mass Cytometry. For the scRNAseq analysis, we enrolled NMBC and MIBC patients and we FACS sorted CD45+ immune cells and CD45- epithelia and stromal cells. Cells were then processed by 3’-based scRNA-seq strategy (10X Genomics) that revealed the transcriptional landscape of sorted populations. Additional patients were enrolled for the Cytomass analysis. Tissue sections were snap frozen, stained and acquired by Hyperion system. The primary endpoint was to unveil differences in the composition and tumor and immune cells in NMIBC and MIBC tumors. RESULTS: The entire cohort consisted of 14 patients with clinically significant bladder cancer who underwent radical cystectomy and TURBT. Our analysis identified differences in immune cells abundance and activation state when comparing NMBC and MIBC samples. Modulations were particularly evident for a subset of PLIN2+ Tumor associated macrophages (TAMs) that was enriched in invasive tumors (Fold change=8.37). In the lymphoid compartment, T regulatory cells (Tregs) showed strong transcriptional changes between NMBC and MIBC samples, that suggest a higher fitness and suppressive function in Tregs that infiltrate MIBC. In addition, our analysis revealed a high heterogeneity in the composition of tumor cells in both cohorts (Figure 1). Finally, Cytomass analysis unveiled modulations in immune cells activation and spatial localization when comparing NMBC and MIBC that will be further investigated. CONCLUSIONS: Our study supports the effectiveness of single cell RNA sequencing in the dissection of the immune landscape in BC and identified immune changes when comparing neoplastic tissue from MIBC with NMBC patients. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e186 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Massimo Lazzeri More articles by this author NicolòMaria Buffi More articles by this author Giovanni Lughezzani More articles by this author Paolo Casale More articles by this author Roberto Hurle More articles by this author Alberto Saita More articles by this author Vittorio Fasulo More articles by this author Marco Paciotti More articles by this author Marta Pandini More articles by this author Roberta Carriero More articles by this author Mattia Carvetta More articles by this author Marta Iovino More articles by this author Piergiuseppe Colombo More articles by this author Grazia Elefante More articles by this author Gianluca Basso More articles by this author Sergio Marchini More articles by this author Paolo Kunderfranco More articles by this author Diletta Di Mitri More articles by this author Expand All Advertisement PDF downloadLoading ...