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FOSL2 deficiency delays nonalcoholic steatohepatitis progression by regulating LY6D-mediated NLRP3 activation

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Abstract

Lymphocyte antigen 6 family member D (LY6D) was enhanced specifically in senescent cells, while its effects on pyroptosis, a programmed cell death, remains unknown. The goal of this study was to assess the role of LY6D in the mediation of pyroptosis during nonalcoholic steatohepatitis (NASH). After screening out LY6D as a specific liver fibrosis-associated gene using the GSE55747 dataset from the GEO database, we established a NASH mouse model using methionine and choline deficient-diet feeding and an in vitro model using lipopolysaccharide (LPS)-treated hepatocytes. LY6D was overexpressed in NASH livers as well as in LPS-treated hepatocytes. Silencing of LY6D inhibited NASH-associated hepatocyte pyroptosis. With the aid of bioinformatics analysis, promoter-luciferase reporter and ChIP-qPCR assays, we identified FOSL2 as an upstream transcription factor of LY6D. FOSL2, which was highly expressed in NASH, promoted LY6D transcription by binding to the promoter of LY6D. Depletion of FOSL2 significantly inhibited NASH-associated hepatocyte pyroptosis, which was significantly reversed after overexpression of LY6D. Moreover, the promotion of hepatocyte pyroptosis by the FOSL2/LY6D axis was significantly attenuated by specific inhibition of NLRP3. These findings suggesting that FOSL2/LY6D axis may be a key molecular axis and a potential target for NASH therapeutics.

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Data availability

The analyzed data sets generated during the study are available from the corresponding author on reasonable request.

Abbreviations

NASH:

Nonalcoholic steatohepatitis

LY6D:

Lymphocyte antigen 6 family member D

LPS:

Lipopolysaccharide

Gsdmd-N:

Gasdermin D N-terminal

α-SMA:

Alpha-smooth muscle actin

AAV:

Adeno-associated virus

MCD:

Methionine and choline deficient

HE:

Hematoxylin–eosin

HRP:

Horseradish peroxidase

ELISA:

Enzyme-linked immunosorbent assay

HBSS:

Hanks' buffered salt solution

ChIP:

Chromatin immunoprecipitation

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Acknowledgements

We thanks to the Natural Science Foundation of Shandong Province (ZR2019BH021), Guangdong Basic and Applied Basic Research Foundation (2019A1515110116 and 2019A1515110221), Natural Science Foundation of Jiangsu Province (BK20200224) and Chinese Foundation for Hepatitis Prevention and Control-TianQing Liver Disease Research Fund Subject (TQGB20200043) for the funding support.

Funding

This work was supported by the Natural Science Foundation of Shandong Province (ZR2019BH021), Guangdong Basic and Applied Basic Research Foundation (2019A1515110116 and 2019A1515110221), Natural Science Foundation of Jiangsu Province (BK20200224) and Chinese Foundation for Hepatitis Prevention and Control-TianQing Liver Disease Research Fund Subject (TQGB20200043) and China International Medical Foundation (Z-2014–06-2102).

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Author notes

  1. Pei-Xin Hu and Mei-Yan Sheng contributed equally to this work.

Authors and Affiliations

  1. Department of Gastroenterology, Shandong Provincial Hospital, Shandong University, No. 324, Jingwuwei 7 Road, Huaiyin District, Jinan, 250021, Shandong Province, People’s Republic of China

    Pei-Xin Hu & Chun-Qing Zhang

  2. Department of Gastroenterology, Jinan Central Hospital, Shandong University, Jinan, 250013, Shandong, People’s Republic of China

    Pei-Xin Hu

  3. Department of Pulmonary and Critical Care Medicine, Shandong Public Health Clinical Center Affiliated to Shandong University, Jinan, 250013, Shandong, People’s Republic of China

    Mei-Yan Sheng

  4. Department of Radiology, Qilu Hospital of Shandong University, No. 107, Wenhua West Road, Lixia District, Jinan, 250012, Shandong Province, People’s Republic of China

    Yan-Ping Liu

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Contributions

PXH and MYS conceived the design of the research and carried out most of the experiments. YPL and CQZ obtained, analyzed and interpreted the data. PXH wrote and revised the manuscript. MYS, YPL and CQZ prepared the figures and provides financial support for all experiments. All authors read and approved the final manuscript.

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Correspondence to Yan-Ping Liu or Chun-Qing Zhang.

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All animal procedures were performed in accordance with the Guidelines for the Care and Use of Laboratory Animals and were approved by the Institutional Ethics Committee of Jinan Central Hospital (Approval number: 2021–08-07–01).

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Hu, PX., Sheng, MY., Liu, YP. et al. FOSL2 deficiency delays nonalcoholic steatohepatitis progression by regulating LY6D-mediated NLRP3 activation. Human Cell 35, 1752–1765 (2022). https://doi.org/10.1007/s13577-022-00760-y

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