Pd51-04 Identification of Novel Hif Independent Oncogenic Targets in Clear Cell Renal Cell Carcinoma

You have accessJournal of UrologyKidney Cancer: Basic Research & Pathophysiology II (PD51)1 May 2024PD51-04 IDENTIFICATION OF NOVEL HIF INDEPENDENT ONCOGENIC TARGETS IN CLEAR CELL RENAL CELL CARCINOMA Jason M. Scovell, Shannon Zayas, Treg Grubb, and Chakraborty Abhishek Jason M. ScovellJason M. Scovell , Shannon ZayasShannon Zayas , Treg GrubbTreg Grubb , and Chakraborty AbhishekChakraborty Abhishek View All Author Informationhttps://doi.org/10.1097/01.JU.0001009360.84490.42.04AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Major improvements have been made in treating advanced renal cell carcinoma (RCC). Unfortunately, the disease is often incurable, underpinning the continued need to establish mechanistic vulnerabilities. We sought to identify dependencies in clear cell RCC (ccRCC), the most common form of RCC, which is driven by pVHL loss. METHODS: H3K27Ac (ChIP-Seq) and mRNA levels (RNASeq) in isogenic ccRCC lines (+/- pVHL) were compared to identify pVHL-dependent transcriptional programs. The functional importance of the differentially-expressed genes was tested in a CRISPR/Cas9 dropout screen in pVHL-deficient ccRCC cells both in vitro and in vivo. Candidate 'hits' were validated using CRISPR/Cas9 tools, rescue experiments, and tumor growth assays in vitro and in vivo. NF-I/A function was evaluated with RNA-Seq and Western blot analysis. RESULTS: Of the ∼200 genes marked by increased H3K27ac and mRNA upregulation in pVHL-null cells, the in vivo CRISPR/Cas9 screen scored ccRCC oncogenes (e.g., SLC2A1), tumor suppressors (e.g., HIF1A), and novel targets e.g., NFIA, which encodes the Nuclear Factor 1A (NF-I/A) transcription factor. Rigorous validations confirmed that NFIA loss causes fitness defects in vitro and in vivo. Interestingly, we found that HIF activation and NF-I/A activation occurred independent of one another in pVHL-deficient cells. Cell based assays and human patient data mined from the Kidney Cancer Renal Cell Carcinoma TCGA (KIRC) cohort established that NFIA loss reduces the expression genes encoding fatty acid metabolism – a key feature of ccRCCs. CONCLUSIONS: We find that NF-I/A is a HIF-independent oncogenic dependency in pVHL-null ccRCCs. NF-I/A transcriptionally regulates fatty acid metabolism, which likely supports it's oncogenic function in ccRCC. Ongoing work seeks to elaborate NFIA's regulatory mechanisms, including NFIA's role as a immune-modulator via production of lipid-derived leukotrienes. Download PPT Source of Funding: Urology Care Foundation Residency Research Award © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e1065 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Jason M. Scovell More articles by this author Shannon Zayas More articles by this author Treg Grubb More articles by this author Chakraborty Abhishek More articles by this author Expand All Advertisement PDF downloadLoading ...