Pd35-03 Genome-Wide Crispr Screens Identified That Dclre1c Regulates the Radiosensitivity of Prostate Cancer

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology III (PD35)1 May 2024PD35-03 GENOME-WIDE CRISPR SCREENS IDENTIFIED THAT DCLRE1C REGULATES THE RADIOSENSITIVITY OF PROSTATE CANCER Toshiki Oka, Koji Hatano, Keisuke Nimura, Yu Ishizuya, Yuki Horibe, Akihiro Yoshimura, Masaru Tani, Yutong Liu, Sassi Nesrine, Yohei Okuda, Akinaru Yamamoto, Toshihiro Uemura, Gaku Yamamichi, Yoshiyuki Yamamoto, Taigo Kato, Atsunari Kawashima, Kazutoshi Fujita, and Norio Nonomura Toshiki OkaToshiki Oka , Koji HatanoKoji Hatano , Keisuke NimuraKeisuke Nimura , Yu IshizuyaYu Ishizuya , Yuki HoribeYuki Horibe , Akihiro YoshimuraAkihiro Yoshimura , Masaru TaniMasaru Tani , Yutong LiuYutong Liu , Sassi NesrineSassi Nesrine , Yohei OkudaYohei Okuda , Akinaru YamamotoAkinaru Yamamoto , Toshihiro UemuraToshihiro Uemura , Gaku YamamichiGaku Yamamichi , Yoshiyuki YamamotoYoshiyuki Yamamoto , Taigo KatoTaigo Kato , Atsunari KawashimaAtsunari Kawashima , Kazutoshi FujitaKazutoshi Fujita , and Norio NonomuraNorio Nonomura View All Author Informationhttps://doi.org/10.1097/01.JU.0001009396.21455.74.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Radiotherapy is widely used for prostate cancer (PCa), but the therapeutic efficacy varies widely among patients. For more effective radiotherapy, it is necessary to elucidate the molecular mechanisms that regulate radiosensitivity. Androgen receptor (AR) is associated with radiosensitivity in PCa, yet the details remain unclear. Here, we used genome-wide CRISPR screens to identify a gene that regulates radiosensitivity in PCa. We further elucidated how the gene regulates radiosensitivity, including its association with AR. METHODS: PCa cell lines, DU145, 22Rv1, and LNCaP, were used to establish CRISPR/Cas9 cell libraries. The CRISPR library contained 90,709 sgRNAs targeting 18,010 genes. The cell libraries were irradiated (4 or 6 Gy) or untreated (control), and cultured for 11 days. The number of each sgRNA was evaluated by sequencing and compared to controls. Enrichment of sgRNA means that the knockout of its target gene enhances radioresistance, and drop-off of sgRNA means that the knockout of its target gene enhances radiosensitivity. RESULTS: CRISPR screens showed that many cell cycle and apoptosis-related genes were enriched, while many DNA repair pathway genes were dropped off. The most potent radiosensitizing target gene, DCLRE1C, was identified by CRISPR screens of three PCa cell lines. Knockout of DCLRE1C significantly increased γH2AX positive cell rate 2.1-fold and comet tail intensity 1.5-fold after irradiation (IR) in DU145 cells. γH2AX and comet tail reflect DNA double-strand breaks and DNA damage, respectively. Knockout of DCLRE1C reduced the radiation dose by 55% required to achieve 90% cell death, but overexpression of DCLRE1C canceled this phenotype. Similar results were confirmed in 22RV1 and PC3 cells. DCLRE1C was activated following IR by Ser-516 phosphorylation. Notably, DCLRE1C phosphorylation was attenuated by an AR signaling inhibitor. CONCLUSIONS: The most potent radiosensitizing target gene, DCLRE1C, was identified by CRISPR screens. The phosphorylation of DCLRE1C plays a pivotal role in AR-mediated radiosensitivity in PCa. Source of Funding: This research was supported by KAKENHI (grant number: 20K18090, 22K09447) © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e725 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Toshiki Oka More articles by this author Koji Hatano More articles by this author Keisuke Nimura More articles by this author Yu Ishizuya More articles by this author Yuki Horibe More articles by this author Akihiro Yoshimura More articles by this author Masaru Tani More articles by this author Yutong Liu More articles by this author Sassi Nesrine More articles by this author Yohei Okuda More articles by this author Akinaru Yamamoto More articles by this author Toshihiro Uemura More articles by this author Gaku Yamamichi More articles by this author Yoshiyuki Yamamoto More articles by this author Taigo Kato More articles by this author Atsunari Kawashima More articles by this author Kazutoshi Fujita More articles by this author Norio Nonomura More articles by this author Expand All Advertisement PDF downloadLoading ...