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GAS5 protects against nonalcoholic fatty liver disease via miR-28a-5p/MARCH7/NLRP3 axis-mediated pyroptosis

Cell Death & Differentiation volume 30, pages 1829–1848 (2023)Cite this article

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Abstract

Nonalcoholic fatty liver disease (NAFLD) is characterised by hepatic steatosis, inflammation, and insulin resistance. The role of long noncoding RNA (lncRNA)-regulated pyroptosis in NAFLD development remains largely unknown. This study aimed to investigate whether NAFLD development is controlled by lncRNA growth-arrest specific transcript 5 (GAS5)/miR-28a-5p/membrane associated ring-CH-type finger 7 (MARCH7)-mediated pyroptosis using in vivo and in vitro models. First, GAS5 expression was decreased but miR-28a-5p expression was increased in the livers of NAFLD patients, high-fat diet (HFD)-fed mice and leptin-deficient obese (Ob/Ob) mice. Furthermore, GAS5 suppressed while miR-28a-5p promoted NAFLD development, and overexpression of miR-28a-5p reversed the GAS5 overexpression-induced attenuation of NAFLD. Mechanistically, GAS5 served as a sponge of miR-28a-5p, and miR-28a-5p enhanced pyroptosis by targeting the 3′ untranslated region (UTR) of the E3 ligase MARCH7 during NAFLD development. MARCH7 interacted with the NOD-like receptor protein 3 (NLRP3) protein, resulting in proteasomal degradation of NLRP3 to inhibit pyroptosis. As expected, MARCH7 knockdown abolished the miR-28a-5p knockdown-induced inhibition of NAFLD development, and the ubiquitin E3 ligase-inactive mutant (W589A/I556A) of MARCH7 failed to inhibit NAFLD development. In conclusion, GAS5 protected against NAFLD development by binding to miR-28a-5p, miR-28a-5p promoted NAFLD development by targeting MARCH7, and MARCH7 ameliorated NAFLD by suppressing NLRP3-mediated pyroptosis. The GAS5/miR-28a-5p/MARCH7/NLRP3 axis plays an important role in NAFLD progression, and it might be a biomarker for NAFLD.

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Fig. 1: GAS5 interacts with miR-28a-5p in hepatic cell lines and the livers of mice.
Fig. 2: Knockdown of miR-28a-5p attenuates HFD-induced hepatic steatosis, inflammation, and insulin resistance in HFD-fed mice.
Fig. 3: Overexpression of miR-28a-5p significantly abolishes GAS5 overexpression-mediated suppression of hepatic steatosis, inflammation, and insulin resistance in Ob/Ob mice.
Fig. 4: miR-28a-5p modulates NLRP3 expression to affect pyroptosis during hepatic steatosis.
Fig. 5: MARCH7 is the direct target of miR-28a-5p that regulates NLRP3 protein expression.
Fig. 6: Interference with MARCH7 expression blocks anti-miR-28a-5p-mediated suppression of hepatic steatosis, inflammation, and insulin resistance in Ob/Ob mice.
Fig. 7: MARCH7 serves as an E3 ligase of NLRP3.
Fig. 8: The ubiquitin E3 ligase-inactive mutant of MARCH7 fails to suppress hepatic steatosis, inflammation, and insulin resistance in Ob/Ob mice.

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Data availability

The original data presented in this study are included in the paper/Supplementary Material, and further inquiries can be directed to the corresponding authors.

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Acknowledgements

We thank Prof. Donghua Yang for constructive discussions.

Funding

This study was supported by the National Natural Science Foundation of China (81602636, 31600269 and 82174292), the Sichuan Science and Technology Program (2019YJ0369 and 2020YJ0401), the China Postdoctoral Science Foundation (2021M693953), the Scientific Research Project of Sichuan Medical Association (S18018), the Postdoctoral Foundation of Jiangsu (2021K385C), Disciplinary Construction Innovation Team Foundation of Chengdu Medical College (No. CMC-XK-2103), Natural Science Foundation of Sichuan Province (2022NSFSC0725) and the Collaborative Innovation Center of Sichuan for Elderly Care and Health (19Z02).

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Authors and Affiliations

  1. Institute of Reproductive Medicine, Medical School, Nantong University, Nantong, China

    Tianxing Chen, Haitao Li & Aiwen Kang

  2. School of Laboratory Medicine, Chengdu Medical College, Chengdu, China

    Yao Meng, Wei Guo, Ke Ren & Wei Zhao

  3. Department of Gastroenterology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, China

    Zhihang Zhou

  4. Department of Infectious Disease, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China

    Lingfeng Wan

  5. Department of Pathology, The First Affiliated Hospital, Zhejiang University, Zhejiang, China

    Xueru Song

  6. Department of Biomedical Sciences and Tung Biomedical Sciences Centre, City University of Hong Kong, Hong Kong, Hong Kong

    Yu Chen

  7. Clinical Laboratory, Clinical Medical College and The First Affiliated Hospital of Chengdu Medical College, Chengdu, China

    Wei Zhao

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Contributions

WZ and TC conceptualised and supervised the overall project. TC, ZZ, HL, LW, AK, WG, KR, and XS performed experiments. ZZ, HL, LW, AK and YC performed validation studies. YC, WG, KR and XS performed formal data analysis. WZ, TC and ZZ drafted the paper, and all authors contributed to the methodology and revisions.

Corresponding author

Correspondence to Wei Zhao.

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The authors declare no competing interests.

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In this study, the clinical specimen collection was approved by the Medical Ethics Committee of the Second Affiliated Hospital of Chongqing Medical University, and all participants provided written informed consent. All animal protocols were approved by the Experimental Animal Ethics Committee of Nantong University.

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Chen, T., Meng, Y., Zhou, Z. et al. GAS5 protects against nonalcoholic fatty liver disease via miR-28a-5p/MARCH7/NLRP3 axis-mediated pyroptosis. Cell Death Differ 30, 1829–1848 (2023). https://doi.org/10.1038/s41418-023-01183-4

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