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The CRL4DCAF6 E3 ligase ubiquitinates CtBP1/2 to induce apoptotic signalling and promote intervertebral disc degeneration

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Abstract

Inflammation and apoptosis are two important pathological causes of intervertebral disc degeneration (IDD). The crosstalk between these two biological processes during IDD pathogenesis remains elusive. Herein, we discovered that chronic inflammation induced apoptosis through a cullin–RING E3 ligase (CRL)-dependent mechanism. Two cullin proteins, CUL4A and 4B, recruited DNA damage-binding protein 1 (DDB1), RING-box protein 1 (RBX1) and DDB1- and CUL4-associated factor 6 (DCAF6) to assemble a CRL4DCAF6 E3 ligase in intervertebral discs (IVDs) derived from IDD patients. The CRL4DCAF6 E3 ligase ubiquitinated and degraded C-terminal-binding protein 1 and 2 (CtBP1/2), two homologues of transcriptional corepressors. The degradation of CtBP1/2 disassociated from the p300–forkhead box O3a (FOXO3a) complex, inducing the expression of B-cell lymphoma 2 (Bcl2)-binding component 3 (BBC3) and causing BBC3-dependent apoptosis. TSC01131, a small molecule that specifically targets CUL4–DDB1 interaction, could inhibit the ubiquitination of CtBP1/2 in vitro and in vivo, thereby decreasing the BBC3 expression level and preventing apoptosis signalling. Using a mouse chronic inflammation model, we found that chronic inflammation could accelerate the IDD process through a conserved CRL4DCAF6-mediated mechanism. The administration of TSC01131 to mice could significantly improve the outcome of IDD. Collectively, our results revealed that inflammation-dependent CRL4DCAF6 E3 ligase triggered apoptosis through the removal of CtBP-mediated transrepression. The blockage of the CRL4DCAF6 E3 ligase by TSC01131 may represent a new therapeutic strategy for IDD treatment.

Key messages

  • CUL4A and CUL4B recruited DDB1, RBX1 and DCAF6 to assemble a CRL4DCAF6 E3 ligase in human IDD biopsies.

  • The CRL4DCAF6 E3 ligase ubiquitinated and degraded CtBP1/2, causing BBC3-dependent apoptosis.

  • A small molecule TSC01131 that specifically targets CUL4–DDB1 interaction could inhibit the ubiquitination of CtBP1/2, improving the outcome of IDD in a mouse model.

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Data availability

All data analysed during this study are included in this published article and its supplementary information files.

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Funding

This study was supported by a grant from Shanghai “Rising Stars of Medical Talent” Youth Development Program–Youth Medical Talents-Specialist Program (2018) (Zhi Chen).

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Author notes

  1. Changchun Tseng and Yingchao Han contributed equally to this work.

Authors and Affiliations

  1. Department of Spine Surgery, Department of Orthopedics, School of Medicine, Renji Hospital, Shanghai Jiao Tong University, NO. 160 Pujian Rd, Pudong District, 200127, Shanghai, China

    Changchun Tseng, Yingchao Han, Zhendong Lv, Qingxin Song, Kun Wang, Hongxing Shen & Zhi Chen

Authors
  1. Changchun Tseng
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  2. Yingchao Han
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  3. Zhendong Lv
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  4. Qingxin Song
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  6. Hongxing Shen
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  7. Zhi Chen
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Contributions

ZC and HS designed the experiments and wrote the manuscript. TC and YH performed the major experiments and data analysis. ZL, QS and KW helped culture cells, generate knockdown cell lines and maintain mice.

Corresponding authors

Correspondence to Hongxing Shen or Zhi Chen.

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Ethics approval and consent to participate

All animal experiments were performed in accordance with a protocol (2018IDD029) reviewed and approved by the ethical board of Renji Hospital, Shanghai Jiao Tong University, Shanghai, China. All the participants signed consent forms reviewed and approved by the ethical board of Shanghai Jiao Tong University, Shanghai, China.

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The authors declare no competing interests.

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Tseng, C., Han, Y., Lv, Z. et al. The CRL4DCAF6 E3 ligase ubiquitinates CtBP1/2 to induce apoptotic signalling and promote intervertebral disc degeneration. J Mol Med 101, 171–181 (2023). https://doi.org/10.1007/s00109-022-02277-1

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  • DOI: https://doi.org/10.1007/s00109-022-02277-1

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