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Epithelial CRL4DCAF2 Is Critical for Maintaining Intestinal Homeostasis Against DSS-Induced Colitis by Regulating the Proliferation and Repair of Intestinal Epithelial Cells

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An Invited Commentary to this article was published on 15 November 2023

Abstract

Background and Aims

Inflammatory bowel disease (IBD) is currently gaining an increasing global interest. Intestinal epithelial barrier dysfunction is crucial toward develo** IBD; however, the underlying mechanisms are not yet elucidated. This study is aimed at elucidating the function of CRL4DCAF2, an E3 ligase, toward mediating intestinal homeostasis.

Methods

Colon samples were collected from patients with IBD and healthy individuals to examine the expression of CRL4DCAF2. CRL4DCAF2 conditional knockdown in mouse intestinal epithelial cells (IECs) (DCAF2EKD) were constructed. DCAF2EKD and their littermate control (DCAF2EWT) were treated with dextran sodium sulfate (DSS) to induce acute colitis. Transcriptome analysis was performed on inflamed colon samples obtained from the mice. Cell cycle regulators were evaluated using real-time polymerase chain reaction (PCR), while tight junction and apoptosis proteins were examined via immunofluorescence and western blot.

Results

CRL4DCAF2 expression was significantly decreased in the inflamed IBD epithelium, and low expression of CRL4DCAF2 associated with high recurrence risk. Mice with DCAF2 specific knockout in IECs suffer from embryonic death. Multiple genes involved in cell proliferation, immune response, and gap junction were differentially expressed in inflamed colon from DCAF2EKD compared with DCAF2EWT. Furthermore, conditional downregulation of CRL4DCAF2 in the intestinal epithelium induced primarily epithelial damage, increased intestinal permeability, and diminished tight junction protein expression. In vivo and in vitro cell transfection experiments revealed that CRL4DCAF2 enhanced cell proliferation by promoting p21 ubiquitination and degradation, thereby inhibiting G2/M cell cycle. In addition, CRL4DCAF2 can also inhibit IEC apoptosis and promote cell autophagy.

Conclusions

CRL4DCAF2 downregulation in IECs promotes intestinal barrier dysfunction and inhibits IEC proliferation, thus making it more susceptible to inflammation.

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Data availability

Sequence data that support the findings of this study are available from the authors and have been deposited in the National Center for Biotechnology Information (NCBI) BioProject with the primary accession code PRJNA791130.

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Acknowledgments

We thank the Life Sciences Institute core facilities, Zhejiang University, for technical assistance. We also thank Prof. ** ** for the mice and expression plasmids. This study was supported by the Zhejiang Provincial Natural Science Foundation of China (LQ21H030010 and LQ21H030013), the National Natural Science Foundation of China (32000628), and the Zhejiang Province Medicine and Health Research Project (2021417815, 2021KY104 and 2023KY804)

Funding

This study was supported by the Zhejiang Provincial Natural Science Foundation of China (LQ21H030010andLQ21H030013), the National Natural Science Foundation of China (32000628), and the Zhejiang Province Medicine and Health Research Project (2021417815, 2021KY104, and 2023KY804).

Author information

Author notes

  1. Yu Zhang, Chaohui Wang, and Lexi Wu contributed equally to this article.

Authors and Affiliations

  1. Department of Gastroenterology, Sir Run Run Shaw Hospital, College of Medicine Zhejiang University, Hangzhou, 310016, China

    Yu Zhang, Lexi Wu, Chenhao Bai, Kaituo Huang, Lingya Yao, Zhou Zhang, Lingna Ye, Rongbei Liu, Mengque Xu, Yuan Zhao & Qian Cao

  2. Inflammatory Bowel Disease Center, Sir Run Run Shaw Hospital, College of Medicine Zhejiang University, Hangzhou, 310016, China

    Yu Zhang, Lexi Wu, Chenhao Bai, Kaituo Huang, Lingya Yao, Zhou Zhang, Lingna Ye, Rongbei Liu, **aolong Ge, Mengque Xu, Yuan Zhao & Qian Cao

  3. Institute of Gastroenterology, Zhejiang University, Hangzhou, 310016, Zhejiang, China

    Yu Zhang, Lexi Wu, Chenhao Bai, Kaituo Huang, Lingya Yao, Zhou Zhang, Lingna Ye, Rongbei Liu, Mengque Xu, Yuan Zhao & Qian Cao

  4. Department of Gastroenterology, Taizhou Central Hospital, Taizhou, 318000, China

    Chaohui Wang

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Contributions

YZ and QC were responsible for conceptualization; YZ, CW, and CB for methodology; KH, LW, LY, ZZ, and RL for investigation; YZ, CW, LW, and QC for writing: original draft; YZ, CW, and QC for writing: review and editing; YZ, CW, and XG for visualization; YZ and QC for supervision; and YZ, RL, MX, and QC for funding acquisition.

Corresponding author

Correspondence to Qian Cao.

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Conflict of interest

The authors declare no competing interests.

Ethical approval and consent to participate

The studies involving human participants were reviewed and approved by the Medical Ethics Committee of Sir Run Run Shaw Hospital (SRRSH) (ID: 2022–0369). The patients provided their written informed consent to participate in this study. All procedures performed in this study involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Declaration of Helsinki and its later amendments. All working stages with animals were conducted following the International Guiding Principles for Biomedical Research Involving Animals in 1985. The study protocol was adhered to (Animal Research: Reporting of In Vivo Experiments (ARRIVE) guideline. All methods were performed in accordance with the relevant guidelines and regulations.

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Zhang, Y., Wang, C., Wu, L. et al. Epithelial CRL4DCAF2 Is Critical for Maintaining Intestinal Homeostasis Against DSS-Induced Colitis by Regulating the Proliferation and Repair of Intestinal Epithelial Cells. Dig Dis Sci 69, 66–80 (2024). https://doi.org/10.1007/s10620-023-08147-1

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