Targeting N4‐acetylcytidine suppresses hepatocellular carcinoma progression by repressing eEF2‐mediated HMGB2 mRNA translation
Abstract Background N4‐acetylcytidine (ac4C) represents a novel messenger RNA (mRNA) modification, and its associated acetyltransferase N‐acetyltransferase 10 (NAT10) plays a crucial role in the initiation and progression of tumors by regulating mRNA functionality. However, its role in hepatocellula...
Saved in:
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2024-09-01
|
| Series: | Cancer Communications |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/cac2.12595 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850268303414525952 |
|---|---|
| author | Hailing Liu Lei Xu Shiwei Yue Hongfei Su Xing Chen Qiumeng Liu Hui Li Huifang Liang Xiaoping Chen Jiefeng He Zeyang Ding Bixiang Zhang |
| author_facet | Hailing Liu Lei Xu Shiwei Yue Hongfei Su Xing Chen Qiumeng Liu Hui Li Huifang Liang Xiaoping Chen Jiefeng He Zeyang Ding Bixiang Zhang |
| author_sort | Hailing Liu |
| collection | DOAJ |
| description | Abstract Background N4‐acetylcytidine (ac4C) represents a novel messenger RNA (mRNA) modification, and its associated acetyltransferase N‐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 pull‐down, 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 HCC mouse 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 library screening revealed Panobinostat as a potent inhibitor of NAT10‐mediated ac4C modification. This inhibition significantly attenuated HCC growth and metastasis in both in vitro experiments using HCC cells and in vivo HCC mouse models. Conclusions Our study identified a novel oncogenic epi‐transcriptome axis involving NAT10‐ac4C/eEF2‐HMGB2, which plays a pivotal role in regulating HCC growth and metastasis. The drug Panobinostat validates the therapeutic potential of targeting this axis for HCC treatment. |
| format | Article |
| id | doaj-art-6514adfb45334e62b419b67a385c48fb |
| institution | OA Journals |
| issn | 2523-3548 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | Wiley |
| record_format | Article |
| series | Cancer Communications |
| spelling | doaj-art-6514adfb45334e62b419b67a385c48fb2025-08-20T01:53:30ZengWileyCancer Communications2523-35482024-09-014491018104110.1002/cac2.12595Targeting N4‐acetylcytidine suppresses hepatocellular carcinoma progression by repressing eEF2‐mediated HMGB2 mRNA translationHailing Liu0Lei Xu1Shiwei Yue2Hongfei Su3Xing Chen4Qiumeng Liu5Hui Li6Huifang Liang7Xiaoping Chen8Jiefeng He9Zeyang Ding10Bixiang Zhang11Hepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei P. R. ChinaHepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei P. R. ChinaHepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei P. R. ChinaHepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei P. R. ChinaDepartment of Hepatopancreatobiliary Surgery Zhejiang Cancer Hospital Hangzhou Institute of Medicine (HIM) Chinese Academy of Sciences Hangzhou Zhejiang P. R. ChinaHepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei P. R. ChinaDepartment of Hepatobiliary Pancreatic Tumor Center Chongqing University Cancer Hospital School of Medicine Chongqing University Chongqing P. R. ChinaHepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei P. R. ChinaHepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei P. R. ChinaDepartment 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. ChinaHepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei P. R. ChinaHepatic Surgery Center Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei P. R. ChinaAbstract Background N4‐acetylcytidine (ac4C) represents a novel messenger RNA (mRNA) modification, and its associated acetyltransferase N‐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 pull‐down, 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 HCC mouse 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 library screening revealed Panobinostat as a potent inhibitor of NAT10‐mediated ac4C modification. This inhibition significantly attenuated HCC growth and metastasis in both in vitro experiments using HCC cells and in vivo HCC mouse models. Conclusions Our study identified a novel oncogenic epi‐transcriptome axis involving NAT10‐ac4C/eEF2‐HMGB2, which plays a pivotal role in regulating HCC growth and metastasis. The drug Panobinostat validates the therapeutic potential of targeting this axis for HCC treatment.https://doi.org/10.1002/cac2.12595N4‐acetylcytidineN‐acetyltransferase 10hepatocellular carcinomamRNA translationtargeted therapy |
| spellingShingle | Hailing Liu Lei Xu Shiwei Yue Hongfei Su Xing Chen Qiumeng Liu Hui Li Huifang Liang Xiaoping Chen Jiefeng He Zeyang Ding Bixiang Zhang Targeting N4‐acetylcytidine suppresses hepatocellular carcinoma progression by repressing eEF2‐mediated HMGB2 mRNA translation Cancer Communications N4‐acetylcytidine N‐acetyltransferase 10 hepatocellular carcinoma mRNA translation targeted therapy |
| title | Targeting N4‐acetylcytidine suppresses hepatocellular carcinoma progression by repressing eEF2‐mediated HMGB2 mRNA translation |
| title_full | Targeting N4‐acetylcytidine suppresses hepatocellular carcinoma progression by repressing eEF2‐mediated HMGB2 mRNA translation |
| title_fullStr | Targeting N4‐acetylcytidine suppresses hepatocellular carcinoma progression by repressing eEF2‐mediated HMGB2 mRNA translation |
| title_full_unstemmed | Targeting N4‐acetylcytidine suppresses hepatocellular carcinoma progression by repressing eEF2‐mediated HMGB2 mRNA translation |
| title_short | Targeting N4‐acetylcytidine suppresses hepatocellular carcinoma progression by repressing eEF2‐mediated HMGB2 mRNA translation |
| title_sort | targeting n4 acetylcytidine suppresses hepatocellular carcinoma progression by repressing eef2 mediated hmgb2 mrna translation |
| topic | N4‐acetylcytidine N‐acetyltransferase 10 hepatocellular carcinoma mRNA translation targeted therapy |
| url | https://doi.org/10.1002/cac2.12595 |
| work_keys_str_mv | AT hailingliu targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT leixu targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT shiweiyue targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT hongfeisu targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT xingchen targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT qiumengliu targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT huili targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT huifangliang targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT xiaopingchen targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT jiefenghe targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT zeyangding targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation AT bixiangzhang targetingn4acetylcytidinesuppresseshepatocellularcarcinomaprogressionbyrepressingeef2mediatedhmgb2mrnatranslation |