Copyright
©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
Adenosine triphosphate-binding pocket inhibitor for mixed lineage kinase domain-like protein attenuated alcoholic liver disease via necroptosis-independent pathway
Han-Ning Xuan Yuan, Hyun Sung Kim, Gye Ryeol Park, Jae Eun Ryu, Ji Eun Kim, In Young Kang, Hye Young Kim, Seung Min Lee, Ju Hee Oh, Eileen L Yoon, Dae Won Jun
Han-Ning Xuan Yuan, Dae Won Jun, Department of Internal Medicine, Hanyang University School of Medicine, Seoul 04763, South Korea
Hyun Sung Kim, Department of Pathology, Hanyang University School of Medicine, Seoul 04763, South Korea
Gye Ryeol Park, Jae Eun Ryu, Ji Eun Kim, In Young Kang, Hye Young Kim, Seung Min Lee, Department of Translational Medicine, Graduate School of Biomedical Science and Engineering, Hanyang University, Seoul 04763, South Korea
Ju Hee Oh, Department of Obstetrics and Gynecology, Institute of Women’s Medical Life Science, Yonsei Cancer Center, Severance Hospital, Yonsei University College of Medicine, Seoul 04763, South Korea
Eileen L Yoon, Department of Internal Medicine, Sanggye Paik Hospital, Inje University College of Medicine, Seoul 01757, South Korea
Co-corresponding authors: Eileen L Yoon and Dae Won Jun.
Author contributions: Yoon EL and Jun DW designed the research study; Kang IY, Ryu JE, Park GR, Oh JH, and Kim JE performed the research; Kang IY analyzed liver organoids; Kim HS analyzed animal model data; Lee SM and Yoon EL contributed to the clarification and explanation of the finding; Xuan Yuan HN wrote the manuscript; All authors have read and approved the final manuscript.
Supported by the National Research Foundation of Korea Grant Funded by the Korea Government, No. RS-2024-00440477; and the Korea Institute of Science and Technology Institutional Program, No. 2E33111-24-042.
Institutional review board statement: The study does not involve human subjects, and therefore, it does not require institutional review board approval.
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 Hanyang University, No. HY-IACUC-2022-0217A and No. HY-IACUC-2023-0276A.
Conflict-of-interest statement: The authors declare that they have no conflict of interest.
Data sharing statement: The data supporting the findings of this study can be requested directly from the corresponding author, at the email address (
noshin@hanyang.ac.kr).
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: Dae Won Jun, PhD, Chief Doctor, Department of Internal Medicine, Hanyang University School of Medicine, 222 Wangsimni-ro, Seongdong-gu, Seoul 04763, South Korea.
noshin@hanyang.ac.kr
Received: May 15, 2024
Revised: October 25, 2024
Accepted: December 11, 2024
Published online: February 14, 2025
Processing time: 240 Days and 1.7 Hours
BACKGROUND
Mixed lineage kinase domain-like protein (MLKL) serves as a critical mediator in necroptosis, a form of regulated cell death linked to various liver diseases. This study aims to specifically investigate the role of MLKL’s adenosine triphosphate (ATP)-binding pocket in facilitating necroptosis-independent pathways that may contribute to liver disease progression. By focusing on this mechanism, we seek to identify potential therapeutic targets that can modulate MLKL activity, offering new strategies for the prevention and treatment of liver-related pathologies.
AIM
To investigate the possibility of using the ATP-binding pocket-associated, necroptosis-independent MLKL pathway as a target for liver diseases.
METHODS
Cell death following necroptosis stimuli was evaluated using cell proliferation assays, flow cytometry, and electron microscopy in various cells. The human liver organoid system was used to evaluate whether the MLKL ATP pocket-binding inhibitor could attenuate inflammation. Additionally, alcoholic and non-alcoholic fatty liver diseases animal models were used to determine whether MLKL ATP pocket inhibitors could attenuate liver injury.
RESULTS
While an MLKL ATP pocket-binding inhibitor did not prevent necroptosis-induced cell death in RAW 264.7 cells, it did reduce the necroptosis-led expression of CXCL2, ICAM, and VCAM. Notably, MLKL ATP pocket inhibitor diminishes the expression of CXCL2, ICAM, and VCAM by inhibiting the IκB kinase and nuclear factor kappa-B pathways without inducing necroptosis-induced cell death in two-dimensional cell culture as well as the human-derived liver organoid system. Although MLKL ATP-binding inhibitor was ineffective in non-alcoholic fatty liver disease animal models, MLKL ATP-binding inhibitor attenuated hepatic inflammation in the alcoholic liver disease model.
CONCLUSION
MLKL ATP pocket-binding inhibitor exerted anti-inflammatory effects through the necroptosis-independent MLKL pathway in an animal model of alcoholic liver disease.
Core Tip: Targeting the adenosine triphosphate-binding pocket of mixed lineage kinase domain-like protein may provide a novel therapeutic strategy for reducing inflammation in alcoholic liver disease, independent of its role in necroptosis, highlighting its potential as a unique target in liver disease management.
