MP65-05 SINGLE-CELL RNA SEQUENCING IDENTIFIES CHANGES IN TUMOR MICROENVIRONMENT INDUCED BY INTERFERON α GENE THERAPY IN MURINE BLADDER CANCER MODEL

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology III (MP65)1 May 2024MP65-05 SINGLE-CELL RNA SEQUENCING IDENTIFIES CHANGES IN TUMOR MICROENVIRONMENT INDUCED BY INTERFERON α GENE THERAPY IN MURINE BLADDER CANCER MODEL Sharada Mokkapati, Alberto Martini, Côme Tholomier, Ganiraju Manyam, Alexis Steinmetz, Yan Chen, and Colin Dinney Sharada MokkapatiSharada Mokkapati , Alberto MartiniAlberto Martini , Côme TholomierCôme Tholomier , Ganiraju ManyamGaniraju Manyam , Alexis SteinmetzAlexis Steinmetz , Yan ChenYan Chen , and Colin DinneyColin Dinney View All Author Informationhttps://doi.org/10.1097/01.JU.0001008756.24343.22.05AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Nadofaragene firadenovec (interferon-α gene therapy; Ad-IFNα) is now approved for BCG-unresponsive bladder cancer (BC). IFNα is a pleiotropic cytokine that causes direct tumor cell killing via TRAIL-mediated apoptosis and antigen processing leading to NK and T-cell activation. To study the effect of Ad-IFNα in a murine BC model, we performed single-cell sequencing on tumor tissue following Ad-IFNα therapy. METHODS: MB49 BC cells were instilled into the bladders of mice; luciferase imaging confirmed tumor growth. Mice were treated with adenovirus control (Ad-Ctrl; no recombinant gene) or Ad-IFNα. At 72 h, bladders were harvested and treated with collagenase/hyaluronidase and TrypLE for cell dissociation. Single cells were suspended in buffer and assessed for viability with Vi-CELL. scRNA seq analysis was done with 10X genomics 3' sequencing. Raw RNAseq data were pre-processed using Cell Ranger single cell software. Seurat was used to normalize and cluster scRNA data. Pooled differential gene expression analysis in specific cell clusters was performed with DESeq2. RESULTS: We identified 15 cell clusters based on marker expression, epithelial, uroplakin-enriched, endothelial, T-cells, neutrophils and macrophage clusters. Top differentially expressed genes between Ad-IFNα and Ad-Ctrl were identified. Within specific cell clusters, IPA analysis revealed significant differences in Ad-IFNα and control. IFN signaling was upregulated in epithelial, endothelial, uroplakin-enriched, lymphocyte and macrophage clusters. Cell death pathways were upregulated in epithelial and endothelial clusters; pyroptosis signaling was enriched in the endothelial cluster. Immunogenic cell death and death receptor signaling were upregulated in epithelial cells. Lymphocytes had upregulated immunogenic cell death and NK cell signaling with a decrease in Th2. Macrophages had hypercytokinemia and upregulated MSP-RON and NOD1/2 signaling and PD1-PD-L1 pathway. Th1, Th2 and macrophage activation pathways were downregulated. CONCLUSIONS: This single cell analysis builds upon our understanding of the impact of Ad-IFNα on tumor cells and the tumor microenvironment. These data will help identify mechanisms to improve patient selection and therapeutic efficacy of Ad-IFNα. Download PPT Source of Funding: This work was funded by DOD grant W81XWH2110698 CA201117 © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e1078 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Sharada Mokkapati More articles by this author Alberto Martini More articles by this author Côme Tholomier More articles by this author Ganiraju Manyam More articles by this author Alexis Steinmetz More articles by this author Yan Chen More articles by this author Colin Dinney More articles by this author Expand All Advertisement PDF downloadLoading ...