PMID- 37534933 OWN - NLM STAT- MEDLINE DCOM- 20230804 LR - 20240212 IS - 2471-254X (Electronic) IS - 2471-254X (Linking) VI - 7 IP - 8 DP - 2023 Aug 1 TI - METTL14 reverses liver fibrosis by inhibiting NOVA2 through an m6A-YTHDF2-dependent mechanism. LID - 10.1097/HC9.0000000000000199 [doi] LID - e0199 AB - BACKGROUND: N6-methyladenosine (m6A), the most prevalent internal RNA modification in eukaryotic cells, is dynamically regulated in response to a wide range of physiological and pathological states. Nonetheless, the involvement of METTL14-induced m6A in liver fibrosis (LF) has yet to be established. METHODS: In vitro, HSC cell lines with knock-down and overexpression of METTL14 were constructed, and the effects of METTL14 gene on the phenotypic function of activated HSCs were observed. The proliferation rate was measured by CCK8 and EDU, the cell proliferation cycle was measured by flow detector, the migration rate was measured by Transwell, and the contractility of F-actin was observed after phalloidin staining. The downstream target gene NOVA2 of METTL14 was screened by combined sequencing of MeRIP-seq and RNA-seq, combined with signal analysis. Adeno-associated virus (AAV) was injected into the tail vein in vivo to knock down the expression of METTL14, so as to further observe the role of METTL14 in the progress of LF. RESULTS: our research showed that the methylase METTL14 content was decreased in hepatic tissue from patients with LF, leading to a lowered degree of m6A modification. Functionally, we discovered that knocking down m6A methyltransferase METTL14 led to increased HSC activation and a substantial worsening of LF. Mechanically, as shown in a multiomics study of HSCs, depleting METTL14 levels decreased m6A deposition onNOVA2 mRNA transcripts, which prompted the activation of YTHDF2 to detect and degrade the decrease of NOVA2 mRNA. CONCLUSIONS: METTL14 functioned as a profibrotic gene by suppressing NOVA2 activity in a mechanism dependent on m6A-YTHDF2. Moreover, knocking down METTL14 exacerbated LF, while NOVA2 prevented its development and partly reversed the damage. CI - Copyright (c) 2023 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the American Association for the Study of Liver Diseases. FAU - Hou, Xiaoxue AU - Hou X AUID- ORCID: 0000-0003-0065-4867 AD - Department of Infectious Disease, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China. FAU - Li, Yuwen AU - Li Y AUID- ORCID: 0009-0008-7673-3987 AD - Department of Pediatrics, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China. FAU - Song, Jiali AU - Song J AUID- ORCID: 0009-0009-1286-0108 AD - Department of Infectious Disease, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China. FAU - Peng, Linya AU - Peng L AUID- ORCID: 0009-0006-0243-0583 AD - Department of Infectious Disease, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China. FAU - Zhang, Wen AU - Zhang W AUID- ORCID: 0000-0002-9474-9359 AD - Department of Infectious Disease, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China. FAU - Liu, Rui AU - Liu R AUID- ORCID: 0000-0002-9962-6980 AD - Department of Tropical Diseases of the Second Affiliated Hospital, Key Laboratory of Tropical Translational Medicine of Ministry of Education, NHC Key Laboratory of Control of Tropical Diseases, School of Tropical Medicine, Hainan Medical University, Haikou, Hainan, China. FAU - Yuan, Hui AU - Yuan H AUID- ORCID: 0000-0001-8533-260 AD - Department of Infectious Disease, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China. FAU - Feng, Tiantong AU - Feng T AUID- ORCID: 0009-0006-0673-0006 AD - Department of Infectious Disease, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China. FAU - Li, Jieying AU - Li J AUID- ORCID: 0009-0000-3981-5694 AD - Department of Tropical Diseases of the Second Affiliated Hospital, Key Laboratory of Tropical Translational Medicine of Ministry of Education, NHC Key Laboratory of Control of Tropical Diseases, School of Tropical Medicine, Hainan Medical University, Haikou, Hainan, China. FAU - Li, Wenting AU - Li W AUID- ORCID: 0000-0001-7734-4838 AD - Department of Tropical Diseases of the Second Affiliated Hospital, Key Laboratory of Tropical Translational Medicine of Ministry of Education, NHC Key Laboratory of Control of Tropical Diseases, School of Tropical Medicine, Hainan Medical University, Haikou, Hainan, China. FAU - Zhu, Chuanlong AU - Zhu C AUID- ORCID: 0000-0001-6557-1654 AD - Department of Infectious Disease, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China. AD - Department of Tropical Diseases of the Second Affiliated Hospital, Key Laboratory of Tropical Translational Medicine of Ministry of Education, NHC Key Laboratory of Control of Tropical Diseases, School of Tropical Medicine, Hainan Medical University, Haikou, Hainan, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20230803 PL - United States TA - Hepatol Commun JT - Hepatology communications JID - 101695860 RN - 0 (Transcription Factors) RN - 0 (Actins) RN - EC 2.1.1.- (Methyltransferases) RN - 0 (NOVA2 protein, human) RN - 0 (Neuro-Oncological Ventral Antigen) RN - 0 (YTHDF2 protein, human) RN - 0 (RNA-Binding Proteins) RN - EC 2.1.1.- (METTL14 protein, human) SB - IM MH - Humans MH - *Transcription Factors MH - *Liver Cirrhosis/genetics MH - Actins MH - Methyltransferases/genetics MH - Neuro-Oncological Ventral Antigen MH - RNA-Binding Proteins/genetics PMC - PMC10409442 COIS- The authors have no conflicts to report. EDAT- 2023/08/03 13:09 MHDA- 2023/08/04 06:42 PMCR- 2023/08/03 CRDT- 2023/08/03 09:33 PHST- 2023/03/30 00:00 [received] PHST- 2023/05/15 00:00 [accepted] PHST- 2023/08/04 06:42 [medline] PHST- 2023/08/03 13:09 [pubmed] PHST- 2023/08/03 09:33 [entrez] PHST- 2023/08/03 00:00 [pmc-release] AID - 02009842-202308010-00017 [pii] AID - HEP4-23-0274 [pii] AID - 10.1097/HC9.0000000000000199 [doi] PST - epublish SO - Hepatol Commun. 2023 Aug 3;7(8):e0199. doi: 10.1097/HC9.0000000000000199. eCollection 2023 Aug 1.