Pd16-02 Increased Spatial Coupling of Integrin and Collagen Iv in the Immunoresistant Clear Cell Renal Cell Carcinoma Tumor Microenvironment

You have accessJournal of UrologyKidney Cancer: Basic Research & Pathophysiology I (PD16)1 May 2024PD16-02 INCREASED SPATIAL COUPLING OF INTEGRIN AND COLLAGEN IV IN THE IMMUNORESISTANT CLEAR CELL RENAL CELL CARCINOMA TUMOR MICROENVIRONMENT Mitchell T. Hayes, Alex Soupir, Taylor Peak, Oscar Ospina, Jonathan Nguyen, Nicholas H. Chakiryan, Natasha Francis, Paola Ramos Echevarria, Kenneth Y. Tsai, Jodi Balasi, Yamila Caraballo Peres, Jasremon Dhillon, Lindsey A. Martinez, Warren E. Gloria, James Mule, Brooke L. Fridley, and Brandon J. Manley Mitchell T. HayesMitchell T. Hayes , Alex SoupirAlex Soupir , Taylor PeakTaylor Peak , Oscar OspinaOscar Ospina , Jonathan NguyenJonathan Nguyen , Nicholas H. ChakiryanNicholas H. Chakiryan , Natasha FrancisNatasha Francis , Paola Ramos EchevarriaPaola Ramos Echevarria , Kenneth Y. TsaiKenneth Y. Tsai , Jodi BalasiJodi Balasi , Yamila Caraballo PeresYamila Caraballo Peres , Jasremon DhillonJasremon Dhillon , Lindsey A. MartinezLindsey A. Martinez , Warren E. GloriaWarren E. Gloria , James MuleJames Mule , Brooke L. FridleyBrooke L. Fridley , and Brandon J. ManleyBrandon J. Manley View All Author Informationhttps://doi.org/10.1097/01.JU.0001009560.23593.56.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Immunotherapy (IO) has improved response rates for patients with advanced clear cell renal cell carcinoma (ccRCC), but most will develop resistance. We sought to utilize cellular-level spatial transcriptomics in the IO naïve and IO exposed settings to better understand IO resistance in ccRCC tumor immune microenvironment (TIME). METHODS: Matched tumor and stromal fields of view (FOV) from primary ccRCC kidney tumors were included for analysis. Spatial molecular imaging (SMI) was obtained for three tissue microarrays using Nanostring's CosMx platform. Cells were phenotyped using Insitutype and unsupervised clustering. Tumor cells were phenotyped using differential gene expression of proximal tubule cells with high VEGF expression. Cell abundance and spatial aggregation by phenotype were then analyzed by treatment status. Spatial gene set enrichment analysis (GSEA) followed by a post hoc spatial analysis of associated transcripts from select enriched gene sets were performed. Global Moran's I test was used to quantify spatial autocorrelation of ligand-receptor (LR) pairs. Multiplex immunofluorescence (mIF) validation testing was performed using antibody markers against proteins from significantly autocorrelated LR pairs. False discovery rate (FDR) p-value adjustment was used for all multiple testing and <0.1 was considered significant. Analysis was completed in R using the spatialTIME and sfdep packages. RESULTS: Seven IO naïve and six IO treated patients were evaluated. IO exposed tumors harbored more stromal CD8+ T cells and neutrophils than IO naïve tumors (FDR<0.001 and FDR=0.011). No spatial aggregation changes were seen following IO. YES1 was the most overexpressed gene in tumor cells (FDR=0.026). On spatial GSEA, the endothelial to mesenchymal (EMT) pathway was enriched and two associated LR transcript pairs were significantly autocorrelated; COL4A1 (gene for collagen IV) and ITGAV (gene for integrin αv-subunit) in the stroma (FDR=0.056). Expression of these genes were highest amongst fibroblasts, tumor cells, and endothelial cells. On mIF validation testing, integrin αv positive cells and cells that were dually positive for integrin αv and smooth muscle actin (a fibroblast marker) were more abundant in the IO exposed stroma compared to IO naïve stroma (p=0.001 and p<0.001). CONCLUSIONS: We found a shift in the abundance of immune cells in the ccRCC TIME following IO treatment. Additionally, we saw significant autocorrelation of two transcripts associated with the EMT pathway, ITGAV and COL4A1. Increased abundance of integrin αv positive cells was confirmed on mIF validation testing. Source of Funding: 2023 Moffitt Team Science-Miles for Moffitt Award, and the Cancer Center Support Grant at the H. Lee Moffitt Cancer Center and Research Institute, an NCI designated Comprehensive Cancer Center (P30 CA 076292) © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e365 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Mitchell T. Hayes More articles by this author Alex Soupir More articles by this author Taylor Peak More articles by this author Oscar Ospina More articles by this author Jonathan Nguyen More articles by this author Nicholas H. Chakiryan More articles by this author Natasha Francis More articles by this author Paola Ramos Echevarria More articles by this author Kenneth Y. Tsai More articles by this author Jodi Balasi More articles by this author Yamila Caraballo Peres More articles by this author Jasremon Dhillon More articles by this author Lindsey A. Martinez More articles by this author Warren E. Gloria More articles by this author James Mule More articles by this author Brooke L. Fridley More articles by this author Brandon J. Manley More articles by this author Expand All Advertisement PDF downloadLoading ...