Abstract 1405: NAT10-mediated RNA acetylation as a critical dependency in pancreatic ductal adenocarcinoma

Abstract Pancreatic ductal adenocarcinoma (PDAC) is the seventh leading cause of cancer-related death worldwide, underscoring the urgent need to uncover its molecular underpinnings and develop novel therapeutic targets. Over 170 RNA modifications constitute the epitranscriptome. Notably, the chemical modification N4-acetylcytidine (ac4C) and its writer enzyme, N-acetyltransferase 10 (NAT10), are overexpressed in most solid tumors, including pancreatic cancer, with high NAT10 expression correlating with worse disease-free survival. Acetylation of mRNA has been shown to enhance translation efficiency and RNA stability, while acetylation of ribosomal RNA is critical for ribosomal biogenesis, thereby influencing protein production. Furthermore, NAT10 has emerged as a critical dependency in most pancreatic cancer cell lines. This study aims to elucidate the role of NAT10 and RNA acetylation in PDAC. By knocking down NAT10 in isogenic pancreatic cell lines, we observed significant reductions in cell proliferation, colony formation, and epithelial-to-mesenchymal transition. Additionally, NAT10-depleted cell lines exhibited increased sensitivity to inhibitors targeting KRAS G12D. RNA sequencing revealed that NAT10 downregulation is associated with upregulated TGF-β signaling, altered lipid metabolism and mitochondrial function. We further mapped ac4C using two orthogonal approaches: ac4C-RNA immunoprecipitation followed by sequencing and a chemical-based method (RetraC:T) that identifies ac4C at base resolution through C:T mismatches. These approaches identified acetylation of oncogenic transcripts implicated in tumorigenesis. Our findings suggest that NAT10 represents a promising therapeutic target in pancreatic ductal adenocarcinoma, offering potential avenues for the development of targeted treatments. Citation Format: Maria Farooq, Ayush Raman, Hamid Beiki, Shalini Oberdoerffer. NAT10-mediated RNA acetylation as a critical dependency in pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1405.