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Knockout of beta-2 microglobulin enhances cardiac repair by modulating exosome imprinting and inhibiting stem cell-induced immune rejection

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Abstract

Background and aims

Allogeneic human umbilical mesenchymal stem cells (alloUMSC) are convenient cell source for stem cell-based therapy. However, immune rejection is a major obstacle for clinical application of alloUMSC for cardiac repair after myocardial infarction (MI). The immune rejection is due to the presence of human leukocyte antigen (HLA) class I molecule which is increased during MI. The aim of this study was to knockout HLA light chain β2-microglobulin (B2M) in UMSC to enhance stem cell engraftment and survival after transplantation.

Methods and results

We developed an innovative strategy using CRISPR/Cas9 to generate UMSC with B2M deletion (B2M–UMSC). AlloUMSC injection induced CD8+ T cell-mediated immune rejection in immune competent rats, whereas no CD8+ T cell-mediated killing against B2M–UMSC was observed even when the cells were treated with IFN-γ. Moreover, we demonstrate that UMSC-derived exosomes can inhibit cardiac fibrosis and restore cardiac function, and exosomes derived from B2M–UMSC are more efficient than those derived from UMSC, indicating that the beneficial effect of exosomes can be enhanced by modulating exosome’s imprinting. Mechanistically, microRNA sequencing identifies miR-24 as a major component of the exosomes from B2M–UMSCs. Bioinformatics analysis identifies Bim as a putative target of miR-24. Loss-of-function studies at the cellular level and gain-of-function approaches in exosomes show that the beneficial effects of B2M–UMSCs are mediated by the exosome/miR-24/Bim pathway.

Conclusion

Our findings demonstrate that modulation of exosome’s imprinting via B2M knockout is an efficient strategy to prevent the immune rejection of alloUMSCs. This study paved the way to the development of new strategies for tissue repair and regeneration without the need for HLA matching.

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Acknowledgement

We thank Dr. IC Bruce for English editing of the manuscript.

Funding

This work was supported by the National Natural Science Foundation of China (NSFC, No. 81870194, No. 91849122, No. 91839101), Jiangsu Province Key Scientific and Technological Project (BE2016669), Suzhou Science and Technology Project (SS201665), Jiangsu Province Peak of Talent in Six Industries (BU24600117), National Natural Science Foundation of China (No. U1601227 to X. Y. Y.), Science and Technology Programs of Guangdong Province (No. 2015B020225006 to X. Y. Y.).

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Authors and Affiliations

  1. Institute for Cardiovascular Science and Department of Cardiovascular Surgery, First Affiliated Hospital of Soochow University, Suzhou, 215123, Jiangsu, People’s Republic of China

    Lianbo Shao, Yu Zhang, Yuqing Zhang, Yanli Wang, Bing Yan, Wenping Xie, Zhenya Shen & Yangxin Li

  2. Department of Cardiac Surgery, Fuwai Hospital, Beijing, 100037, People’s Republic of China

    Xiangbin Pan

  3. Department of Cardiology, The First Hospital of Jilin University, Changchun, 130041, Jilin, People’s Republic of China

    Bin Liu

  4. Department of Cardiology, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, People’s Republic of China

    Chun Liang

  5. Fuwai Yunnan Cardiovascular Hospital, Kunming, 650302, Yunnan, People’s Republic of China

    Yi Sun

  6. Guangzhou Medical University, Guangzhou, 510080, Guangdong, People’s Republic of China

    Xi-Yong Yu

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  1. Lianbo Shao
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  2. Yu Zhang
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Contributions

YL conceived, designed the study, analyzed data and wrote the manuscript. LS, YZ, YZ, YW, BY and WX performed the experiments and collected data. CL, BL, XP, YS, ZS and XY interpreted the data and revised the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Yangxin Li.

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Research involving animal participants

The animal experiments were approved by the Animal Care and Use Committee of Soochow University.

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The authors declare no conflicts of interest.

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Lianbo Shao, Yu Zhang, Xiangbin Pan, Bin Liu and Chun Liang have contributed equally to this work.

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Shao, L., Zhang, Y., Pan, X. et al. Knockout of beta-2 microglobulin enhances cardiac repair by modulating exosome imprinting and inhibiting stem cell-induced immune rejection. Cell. Mol. Life Sci. 77, 937–952 (2020). https://doi.org/10.1007/s00018-019-03220-3

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