Basic Study
Copyright ©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Mar 28, 2024; 30(12): 1764-1776
Published online Mar 28, 2024. doi: 10.3748/wjg.v30.i12.1764
ALKBH5 suppresses autophagic flux via N6-methyladenosine demethylation of ZKSCAN3 mRNA in acute pancreatitis
Tao Zhang, Shuai Zhu, Geng-Wen Huang
Tao Zhang, Shuai Zhu, Geng-Wen Huang, Department of General Surgery, Xiangya Hospital, Central South University, Changsha 410005, Hunan, China.
Tao Zhang, Shuai Zhu, Geng-Wen Huang, National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha 410005, Hunan, China.
Co-corresponding authors: Shuai Zhu and Geng-Wen Huang.
Author contributions: Zhu S and Huang GW conceived, designed and refined the study protocol; Zhang T finished the experiments, collected and analyzed the data and drafted the manuscript; Zhu S and Huang GW reviewed and revised the manuscript. All authors had access to the study data and reviewed and approved the final manuscript. Zhu S and Huang GW contributed equally to this work as co-corresponding authors. The reasons for designating Zhu S and Huang GW as co-corresponding authors are twofold. First, co-corresponding authors jointly conceived the overall design of the study and revised the manuscript. Second, they jointly provided financial support for the study. In summary, we believe that designating Zhu S and Huang GW as co-corresponding authors accurately reflects our team's collaborative spirit, equal contributions, and diversity.
Supported by National Natural Science Foundation of China, No. 81802450; and Natural Science Foundation of Hunan Province, No. 2020JJ4133 and No. 2021JJ31135.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board at Xiangya Hospital of Central South University.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Xiangya Hospital of Central South University.
Conflict-of-interest statement: The authors declare that they have no conflicts of interest.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Shuai Zhu, MD, PhD, Surgeon, Department of Pancreatic Surgery, Xiangya Hospital, Central South University, No. 87 Xiangya Road, Changsha 410005, Hunan Province, China. zhushuai@csu.edu.cn
Received: November 11, 2023
Peer-review started: November 11, 2023
First decision: January 24, 2024
Revised: February 3, 2024
Accepted: March 6, 2024
Article in press: March 6, 2024
Published online: March 28, 2024
Abstract
BACKGROUND

Increasing evidence has demonstrated that N6-methyladenosine (m6A) RNA modification plays an essential role in a wide range of pathological conditions. Impaired autophagy is a critical hallmark of acute pancreatitis (AP).

AIM

To explore the role of the m6A modification of ZKSCAN3 in the regulation of autophagy in AP.

METHODS

The AP mouse cell model was established by cerulein-treated mouse pancreatic acinar cells (MPC-83), and the results were confirmed by the levels of amylase and inflammatory factors. Autophagy activity was evaluated by specific identification of the autophagy-related microstructure and the expression of autophagy-related genes. ZKSCAN3 and ALKBH5 were knocked down to study the function in AP. A m6A RNA binding protein immunoprecipitation assay was used to study how the m6A modification of ZKSCAN3 mRNA is regulated by ALKBH.

RESULTS

The increased expression of amylase and inflammatory factors in the supernatant and the accumulation of autophagic vacuoles verified that the AP mouse cell model was established. The downregulation of LAMP2 and upregulation of LC3-II/I and SQSTM1 demonstrated that autophagy was impaired in AP. The expression of ZKSCAN3 was upregulated in AP. Inhibition of ZKSCAN3 increased the expression of LAMP2 and decreased the expression of the inflammatory factors, LC3-II/I and SQSTM1. Furthermore, ALKBH5 was upregulated in AP. Knockdown of ALKBH5 downregulated ZKSCAN3 expression and restored decreased autophagic flux in AP. Notably, the bioinformatic analysis revealed 23 potential m6A modification sites on ZKSCAN3 mRNA. The m6A modification of ZKSCAN3 mRNA was significantly decreased in AP. Knockdown of ALKBH5 increased the modification of ZKSCAN3 mRNA, which confirmed that ALKBH5 upregulated ZKSCAN3 expression in a m6A-dependent manner.

CONCLUSION

ALKBH5 inhibits autophagic flux through m6A demethylation of ZKSCAN3 mRNA in AP, thereby aggravating the severity of the disease.

Keywords: Acute pancreatitis, Autophagy, ZKSCAN3, N6-methyladenosine, ALKBH5

Core Tip: Acute pancreatitis (AP) is a common emergency in digestive system. Impaired autophagy is one of important pathogenic mechanisms of AP, however, its regulatory mechanism remains unclear. N6-methyladenosine modification and ZKSCAN3 are crucial regulatory factors of autophagy, but their roles in AP are not well-defined. This study confirmed that the demethylase ALKBH5 can inhibit autophagy flux by upregulating ZKSCAN3, thereby exacerbating the inflammatory severity of AP. The findings of this study provided new insights into the autophagy regulation mechanism and offered a novel direction for early intervention in AP.