MP66-04 THE ROLE OF CHROMATIN AND HISTONE SUCCINYLATION AS A POST-TRANSLATIONAL MODIFICATION IN NON-MUSCLE INVASIVE BLADDER CANCER

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology III (MP66)1 Sep 2021MP66-04 THE ROLE OF CHROMATIN AND HISTONE SUCCINYLATION AS A POST-TRANSLATIONAL MODIFICATION IN NON-MUSCLE INVASIVE BLADDER CANCER Genesis Rivera-Marquez, Beatriz Walter, Maria Merino, and Vladimir Valera Genesis Rivera-MarquezGenesis Rivera-Marquez More articles by this author , Beatriz WalterBeatriz Walter More articles by this author , Maria MerinoMaria Merino More articles by this author , and Vladimir ValeraVladimir Valera More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002106.04AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Bladder cancer has been characterized as one of the most genomically unstable cancers. Epigenetic events such as DNA methylation and histone acetylation have been described to play key roles in the disease onset and progression. These events are controlled ultimately by post-translational modifications (PTM) of proteins. In this study, we aimed to characterize a recently described PTM such as protein succinylation and its role in non-muscle invasive bladder cancer pathogenesis. METHODS: Well-characterized cell lines representing non-muscle invasive bladder cancer including MGH-U3 (pTa/pT1-derived), T24 (pTa-derived), RT4 (pT1-derived), RT-112 (pTa) were analyzed. Additionally, two muscle-invasive cancer derived cell lines UM-UC3 and SW-780 were included for comparison. All analysis were normalized to the reference Primary Bladder Epithelial cells (untransformed). Cell compartment fractionation was achieved by centrifugation under detergent, and included cell membrane, cytoplasmic, nuclear soluble and chromatin fractions. Extracts were then probed with Anti-Pan-Succinyl Lysine antibody. Histone-3 Acetylation and methylation were also tested. Chromatin immunoprecipitation with pan-succinyl-lysine was then performed and genomic DNA sent for Next Generation Sequencing (Chip-Seq) to identify transcription factors bound to succinylated chromatin. RESULTS: All cell compartments contained succinylated proteins, predominantly the cytosolic and chromatin/histone fractions. Histone-3 succinylation was prominent in all cancer cell lines predominantly the NMIBC ones. However, there was no clear association between succinylation and p53 and/or FGFR3 mutation status. Chip-seq analysis demonstrated that most of the represented marks on sequencing mapped to intergenic regions (>50%) and were associated with euchromatin markers, i.e, associated with gene activation. Metabolomic profiling of the cells also varied, ranging from mostly glycolytic pattern to predominatly ox-phos dependent. Accumulation of TCA-intermediaries such as Acetyl-Co-A, Fumarate, Citrate, and Succinate was marked in the NMIBC cells. CONCLUSIONS: Our results suggests that succinylation of chromatin components may be important for regulating nuclear functions, including chromatin dynamics and transcription and therefore in the pathogenesis of non-muscle invasive bladder cancer. Source of Funding: NIH Intramural Research Program © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e1130-e1130 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Genesis Rivera-Marquez More articles by this author Beatriz Walter More articles by this author Maria Merino More articles by this author Vladimir Valera More articles by this author Expand All Advertisement Loading ...