Mp65-11 Arid1a Loss in Bladder Cancer Correlates With Genetic Signatures of Cell Migration Dysregulation

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology III (MP65)1 May 2024MP65-11 ARID1A LOSS IN BLADDER CANCER CORRELATES WITH GENETIC SIGNATURES OF CELL MIGRATION DYSREGULATION Vincent D'Andrea, Christopher Magnani, Timothy Hanlon, Rea Chroneos, Timothy Clinton, Filipe L. F. Carvalho, and Kent Mouw Vincent D'AndreaVincent D'Andrea , Christopher MagnaniChristopher Magnani , Timothy HanlonTimothy Hanlon , Rea ChroneosRea Chroneos , Timothy ClintonTimothy Clinton , Filipe L. F. CarvalhoFilipe L. F. Carvalho , and Kent MouwKent Mouw View All Author Informationhttps://doi.org/10.1097/01.JU.0001008756.24343.22.11AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: ARID1A is an essential component of the large chromatin-remodeling BAF complex. 20% of bladder cancers (BC) contain genomic alterations in ARID1A; however, the biological consequences of ARID1A loss in BC remain to be fully elucidated. Based on the observation that the frequency of ARID1A alterations increases with increasing stage of disease, we hypothesize that ARID1A deficiency primes an invasive transformation in BC. We show that ARIDA1 loss potentiates an invasive phenotype in BC cell lines and correlates with genetic signatures of cell migration dysregulation, providing evidence for its transformative role in the development of higher stage and metastatic BC. METHODS: CRISPR/Cas9-mediated gene editing was used to delete ARID1A in three cell lines with wild-type (WT) ARID1A (T24, SW1710, and 5637). Loss of ARID1A protein was confirmed by immunoblot. Invasion studies (scratch assay) were performed on the knockout (KO) and WT cell lines. Whole transcriptome bulk mRNA sequencing (RNA-seq) was performed via Illumina NovaSeq platform with alignment in the Salmon-based nf-core pipeline. Differential gene expression (DGE) analysis was performed using DESeq2. Gene Set Enrichment Analysis (GSEA) linked to the Molecular Signature Database (MSigDB) further characterized biological pathways altered in KOs with gene ontology (GO) analysis. RESULTS: KO of ARID1A yielded ARID1A-deficient clones for SW1710, T24, and 5637. A scratch assay was performed with the cell lines in triplicate, with each ARID1A-deficient cell line showing a trend towards more rapid wound closure than its matched WT cell line (WT=31.5% vs. KO=53.5% for SW1710, p=0.15; WT=17.2% vs. KO=52.0% for T24, p=0.07; WT=10.4% vs. KO=13.9% for 5637, p=0.59) (Figure 1A). GO analysis matrix derived from RNA-seq showed gene expression dysregulation in cell migration and motility. CONCLUSIONS: To explore the increased frequency of ARID1A alterations in metastatic BC compared to non-invasive tumors, we developed ARID1A-deficient BC cell lines which showed a trend towards a more invasive phenotype following ARID1A KO. These trends were coupled with gene expression profiles which correlated with cell migration and adhesion dysregulation. Taken together, these results suggest that ARID1A deficiency in BC may prime tumors for a more invasive phenotype. Download PPT Source of Funding: BCAN Young Investigator Award, Karin Grunebaum Cancer Research Foundation © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e1081 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Vincent D'Andrea More articles by this author Christopher Magnani More articles by this author Timothy Hanlon More articles by this author Rea Chroneos More articles by this author Timothy Clinton More articles by this author Filipe L. F. Carvalho More articles by this author Kent Mouw More articles by this author Expand All Advertisement PDF downloadLoading ...