PD25-12 IDENTIFICATION OF INTRATUMOR PATHOGENS FROM SINGLE-CELL RNA SEQUENCING, CELLULAR DYSFUNCTION AND IMMUNE RESPONSE

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You have accessJournal of UrologyCME1 Apr 2023PD25-12 IDENTIFICATION OF INTRATUMOR PATHOGENS FROM SINGLE-CELL RNA SEQUENCING, CELLULAR DYSFUNCTION AND IMMUNE RESPONSE Laura Bukavina, Mohit Sindhani, Henkel Valentine, Daniel Ranti, Kirtishri Mishra, Alexander Kutikov, Shilpa Gupta, John Sfakianos, Mahmoud Ghannoum, Mauricio Retuerto, Andres Correa, Robert Uzzo, and Philip Abbosh Laura BukavinaLaura Bukavina More articles by this author , Mohit SindhaniMohit Sindhani More articles by this author , Henkel ValentineHenkel Valentine More articles by this author , Daniel RantiDaniel Ranti More articles by this author , Kirtishri MishraKirtishri Mishra More articles by this author , Alexander KutikovAlexander Kutikov More articles by this author , Shilpa GuptaShilpa Gupta More articles by this author , John SfakianosJohn Sfakianos More articles by this author , Mahmoud GhannoumMahmoud Ghannoum More articles by this author , Mauricio RetuertoMauricio Retuerto More articles by this author , Andres CorreaAndres Correa More articles by this author , Robert UzzoRobert Uzzo More articles by this author , and Philip AbboshPhilip Abbosh More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003303.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Pathogenic microbes can induce cellular dysfunction and alters immune response to therapy. Taking advantage of single-cell RNA-sequencing (scRNA-seq), we aim to assess the transcriptomic features of the single cell level to identify intracellular pathogens in patients with high grade bladder cancer. METHODS: CellRanger and Alevin were utilized to extract cell barcodes and unique molecular identifiers. Low complexity reads <20 was considered negative, and PCR duplicates were removed. Seuret Findmarkers, scIntegrate, and scDensity Plot were utilized to normalize the data, cluster the cells, and calculate markers in each cluster of differentially expressed genes between the two conditions. We focused our analysis on Pseudomonas aeruginosa due to high urogenital virulence, and immunomodulation capabilities by pattern recognition receptors and inflammasome. RESULTS: Patients with history of high-grade BC demonstrated increased presence of Aeromonas (175,473 reads, 54%), Pseudomonas (75,002 reads, 26.73%), and Bacillus (15,410 reads, 5.49%) in their tissue samples, not visualized in healthy controls (counts <10). Among 8,324 cells (across 7 patients), classical CD14+ monocyte subtype was most likely to be exposed to Pseudomonas species (4150 /5069, 81.8%), followed by non-classical CD16+ monocytes (722/1040, 69.4%), and CD8+ Tcells (348/680, 51.1%). (Figure 1A-C). Relative gene expression level between exposed and non-exposed monocytes demonstrated total 25 upregulated genes, with increased expression GNBL21 (aka: RACK1, log10 4.1), MALAT1(log10 3.7), TCEB2(log10 2.9) (increasing proliferation, migration, and reduction of cell autophagy) compared to non-infected, and normal controls. Pseudomonas involvement of non-monocyte cells (CD8+, CD4+Th17, CD4+ naïve) was associated with disease progression (2/5 patients), as well as increased expression of IFI6/27(interferon alpha inducible protein 6 & 27), ITAC (interferon inducible T cell alpha-chemoattractant). CONCLUSIONS: Altogether, our analysis depicted the distribution of intracellular bacteria not previously recognized within tumor microenvironment utilizing scRNAseq, and revealed overexpression of RACK1 in Pseudomonas positive cells, associated with apoptosis resistance. Source of Funding: Bladder Cancer Advocacy Network Young Investigator Award © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e734 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Laura Bukavina More articles by this author Mohit Sindhani More articles by this author Henkel Valentine More articles by this author Daniel Ranti More articles by this author Kirtishri Mishra More articles by this author Alexander Kutikov More articles by this author Shilpa Gupta More articles by this author John Sfakianos More articles by this author Mahmoud Ghannoum More articles by this author Mauricio Retuerto More articles by this author Andres Correa More articles by this author Robert Uzzo More articles by this author Philip Abbosh More articles by this author Expand All Advertisement PDF downloadLoading ...