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N6-methyladenosine hypomethylation of circGPATCH2L regulates DNA damage and apoptosis through TRIM28 in intervertebral disc degeneration

Cell Death & Differentiation volume 30, pages 1957–1972 (2023)Cite this article

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Abstract

Circular RNAs (circRNAs) are a class of noncoding RNAs that have been found to be involved in intervertebral disc degeneration (IVDD) progression, and N6-methyladenosine (m6A) broadly exists in circRNAs. Here, we identified circGPATCH2L with a low m6A methylation level to be upregulated in degenerative nucleus pulposus tissues. Mechanistically, as a protein decoy for tripartite motif containing 28 (TRIM28) within aa 402–452 region, circGPATCH2L abrogates the phosphorylation of TRIM28 and inhibits P53 degradation, which contributes to DNA damage accumulation and cellular apoptosis and leads to IVDD progression. Moreover, m6A-methylated circGPATCH2L is recognised and endoribonucleolytically cleaved by a YTHDF2-RPL10-RNase P/MRP complex to maintain the physiological state of nucleus pulposus cells. Thus, our data show the physiological significance of m6A modification in regulating circRNA abundance and provide a potentially effective therapeutic target for the treatment of IVDD.

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Fig. 1: circGPATCH2L is hypomethylated and upregulated in IVDD.
Fig. 2: Characterisation of circGPATCH2L and its function in NPCs.
Fig. 3: circGPATCH2L promotes DNA damage and apoptosis in NPCs through TRIM28.
Fig. 4: circGPATCH2L functions in NPCs by binding the aa 402–452 region of TRIM28.
Fig. 5: m6A methylated circGPATCH2L was subject to endoribonucleolytic cleavage via YTHDF2-RPL10-RNase P/MRP complex.
Fig. 6: Knockdown of circGPATCH2L in NPCs represses IVDD in a mouse model.
Fig. 7: Schematic illustration for the working model of circGPATCH2L.

Data availability

The raw data from the m6A-circRNA epitranscriptomic microarray of NPCs have been deposited in the Gene Expression Omnibus database under the accession code (GSE199993). All of the full-length original western blots are provided as supplementary material. Data supporting the present study are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank all voluntary intervertebral disc donors. We are grateful to the Central-Lab of Huashan Hospital for their excellent technical help.

Funding

This work was supported by grants from the National Natural Science Foundation of China (Numbers 82172490, 82102620, 81972109, 82072488) and Shanghai Science and Technology Development Funds (Numbers 23YF1404200).

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  1. These authors contributed equally: Zhenhao Chen, Jian Song, Lin Xie.

Authors and Affiliations

  1. Department of Orthopedics, Huashan Hospital, Fudan University, Shanghai, 200000, China

    Zhenhao Chen, Jian Song, Guangyu Xu, Chaojun Zheng, Xinlei Xia, Feizhou Lu, Xiaosheng Ma, Fei Zou, Jianyuan Jiang & Hongli Wang

  2. Department of Orthopedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430000, China

    Lin Xie

  3. Department of Orthopedics, Shanghai Fifth People’s Hospital, Fudan University, Shanghai, 200000, China

    Feizhou Lu

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Contributions

HW, JY, and ZC designed the experiments. ZC, JS, and LX conducted the majority of the assays, acquired the data and wrote the manuscript. FZ, LX, and SM performed the animal experiments. ZC, JS, LX, and FZ analysed the data. FL, XX, GX, and CZ collected human samples and provided technical assistance. HW, JY, and FZ arranged the results and revised the manuscript.

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Correspondence to Fei Zou, Jianyuan Jiang or Hongli Wang.

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The study protocols were approved by the Ethics Committee of Fudan University, Huashan Hospital, and informed consent was obtained from each donor. The Animal Care and Use Committee of Fudan University approved the surgical interventions, treatments, and postoperative animal care procedures in this study.

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Chen, Z., Song, J., Xie, L. et al. N6-methyladenosine hypomethylation of circGPATCH2L regulates DNA damage and apoptosis through TRIM28 in intervertebral disc degeneration. Cell Death Differ 30, 1957–1972 (2023). https://doi.org/10.1038/s41418-023-01190-5

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