Targeting N4-acetylcytidine suppresses hepatocellular carcinoma progression by repressing eEF2-mediated HMGB2 mRNA translation  

作　　者：Hailing Liu Lei Xu Shiwei Yue Hongfei Su Xing Chen Qiumeng Liu Hui Li Huifang Liang Xiaoping Chen Jiefeng He Zeyang Ding Bixiang Zhang 

机构地区：[1]Hepatic Surgery Center,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei,P.R.China [2]Hubei Key Laboratory of Hepato‑Pancreatic‑Biliary Diseases,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei,P.R.China [3]Department of Hepatopancreatobiliary Surgery,Zhejiang Cancer Hospital,Hangzhou Institute of Medicine(HIM),Chinese Academy of Sciences,Hangzhou,Zhejiang,P.R.China [4]Department of Hepatobiliary Pancreatic Tumor Center,Chongqing University Cancer Hospital,School of Medicine,Chongqing University,Chongqing,P.R.China [5]Key Laboratory of Organ Transplantation,Ministry of Education,Wuhan,Hubei,P.R.China [6]Department of Hepatobiliary Surgery,Key Laboratory of Hepatobiliary and Pancreatic Diseases of Shanxi Province(Preparatory),Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences,Shanxi Medical University,Taiyuan,Shanxi,P.R.China

出　　处：《Cancer Communications》2024年第9期1018-1041,共24页癌症通讯（英文）

基　　金：State Key Project on Infection Disease of China,Grant/Award Number:2018ZX10723204-003-003;Tongji Hospital(HUST)Foundation for Excellent Young Scientist,Grant/Award Number:2020YQ05;National Basic Research Program of China,Grant/Award Number:2020YFA0710700;Knowledge Innovation Program ofWuhan-Shuguang Project,Grant/Award Number:2022020801020456;the first level of the public health youth top talent project of Hubei province,Grant/Award Number:2022SCZ051;National Natural Science Foundation of China,Grant/Award Numbers:81874065,81874189,82203823,82273441。

摘　　要：Background:N4-acetylcytidine(ac4C)represents a novel messenger RNA(mRNA)modification,and its associated acetyltransferaseN-acetyltransferase 10(NAT10)plays a crucial role in the initiation and progression of tumors by regulating mRNA functionality.However,its role in hepatocellular carcinoma(HCC)development and prognosis is largely unknown.This study aimed to elucidate the role of NAT10-mediated ac4C in HCC progression and provide a promising therapeutic approach.Methods:The ac4C levels were evaluated by dot blot and ultra-performance liquid chromatography-tandem mass spectrometry with harvested HCC tissues.The expression of NAT10 was investigated using quantitative real-time polymerase chain reaction,western blotting,and immunohistochemical staining across 91 cohorts of HCC patients.To explore the underlying mechanisms of NAT10-ac4C in HCC,we employed a comprehensive approach integrating acetylated RNA immunoprecipitation and sequencing,RNA sequencing and ribosome profiling analyses,along with RNA immunoprecipitation,RNA pulldown,mass spectrometry,and site-specific mutation analyses.The drug affinity responsive targets stability,cellular thermal shift assay,and surface plasmon resonance assays were performed to assess the specific binding of NAT10 and Panobinostat.Furthermore,the efficacy of targeting NAT10-ac4C for HCC treatment was elucidated through in vitro experiments using HCC cells and in vivo HCCmouse models.Results:Our investigation revealed a significant increase in both the ac4C RNA level and NAT10 expression in HCC.Notably,elevated NAT10 expression was associated with poor outcomes in HCC patients.Functionally,silencing NAT10 suppressed HCC proliferation and metastasis in vitro and in vivo.Mechanistically,NAT10 stimulates the ac4C modification within the coding sequence(CDS)of high mobility group protein B2(HMGB2),which subsequently enhances HMGB2 translation by facilitating eukaryotic elongation factor 2(eEF2)binding to the ac4C sites on HMGB2 mRNA’s CDS.Additionally,high-throughput compound libra

关 键 词：N4-acetylcytidine N-acetyltransferase 10 hepatocellular carcinoma mRNA translation targeted therapy 

分 类 号：R735.7[医药卫生—肿瘤]