Abstract
Mesenchymal stem cells (MSCs) are multipotent stem cells that can exert immunomodulatory capacity upon stimulation with pro-inflammatory cytokines. Our previous work has identified Cullin 4B (CUL4B), a scaffold protein in the CUL4B-RING E3 ligase (CRL4B) complex, as a key regulator in the differentiation of MSCs. Here, we demonstrate the critical role of CUL4B in regulating the immunosuppressive function of MSCs. When stimulated with pro-inflammatory cytokines, MSCs lacking CUL4B display enhanced immunosuppressive capacity, which is mediated by the elevated inducible nitric oxide synthase (iNOS). TGF-β signaling can suppress iNOS by inhibiting its transcription as well as promoting its protein degradation. We show that the CRL4B complex cooperates with PRC2 complex and HDACs to repress transcription of Dlx1 and Pmepa1, two inhibitors of TGF-β signaling, leading to decreased expression and accelerated degradation of iNOS. Our study unveils the CRL4B complex as a potential therapeutic target in promoting the immunosuppressive capacity of MSCs.
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Data availability
The raw data for RNA-seq and ChIP-seq can be assessed at PRJNA846938. The uncropped Western blots are provided as Supplementary material. All the other raw data supporting the findings of this study are available from the corresponding authors upon request.
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Acknowledgements
We thank Translational Medicine Core Facility of Shandong University and the School of Basic Medical Sciences Core Facility of Shandong University for technical support.
Funding
This work was supported by National Key R&D Program of China (2022YFC2703700, 2022YFC2703701 to YG and 2022YFC2703700, 2022YFC2703703 to GS), National Natural Science Foundation of China (31872810 to YG and 32200483 to YS), Taishan Youth Scholar (tsqn202312025 to GS) and Shandong Provincial Natural Science Foundation (ZR2021QH284 to YS).
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GS and YG performed study concept and design. RY performed most of the cell and molecular experiments and analyzed the RNA-seq/ChIP-seq data. HH helped to perform the mouse experiments. SC and LQ helped to perform mouse experiments. MD helped to set up experiments. YM helped to perform cell experiments. YW, WJ and YS helped to perform molecular experiments. YZ, MW, QL and BJ provided acquisition, analysis and interpretation of the data. RY, YG and GS wrote and revised the manuscript. All the authors read and approved the final paper.
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This study did not involve any human participants. All the mouse experiments were performed in accordance with the guidelines from the Association for Assessment and Accreditation of Laboratory Animal Care and approved by the Animal Care and Use Committee of School of Basic Medical Sciences of Shandong University (No. ECSBMSSDU2018-2-004).
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Yu, R., Han, H., Chu, S. et al. Cullin 4B-RING E3 ligase negatively regulates the immunosuppressive capacity of mesenchymal stem cells by suppressing iNOS. Cell Death Differ 32, 149–161 (2025). https://doi.org/10.1038/s41418-024-01359-6
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DOI: https://doi.org/10.1038/s41418-024-01359-6
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