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Runx1 Deficiency Promotes M2 Macrophage Polarization Through Enhancing STAT6 Phosphorylation

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Abstract

Our previous study had demonstrated that Runx1 promoted LPS-induced macrophage inflammatory response, however, the role of Runx1 in M2 macrophage polarization still remains largely unknown. This study was conducted to investigate the role of Runx1 in IL-4/IL-13-induced M2 macrophage polarization and its potential regulatory mechanism. We found that exposure of macrophages to IL-4/IL-13 induced a remarkable increasement in Runx1 expression level. Specifically, we established genetically modified mice lacking Runx1 in myeloid cells, including macrophages. RNA-Seq was performed to identify differentially expressed genes (DEGs) between Runx1 knockout and WT control bone marrow-derived macrophages (BMDMs). We identified 686 DEGs, including many genes which were highly expressed in M2 macrophage. In addition, bioinformatics analysis indicated that these DEGs were significantly enriched in extracellular matrix-related processes. Moreover, RT-qPCR analysis showed that there was an obvious upregulation in the relative expression levels of M2 marker genes, including Arg1, Ym1, Fizz1, CD71, Mmp9, and Tgm2, in Runx1 knockout macrophages, as compared to WT controls. Consistently, similar results were obtained in the protein and enzymatic activity levels of Arg1. Finally, we found that the STAT6 phosphorylation level was significantly enhanced in Runx1 knockout macrophages, and the STAT6 inhibitor AS1517499 partly reduced the upregulated effect of Runx1 deficiency on the M2 macrophage polarization. Taken together, Runx1 deficiency facilitates IL-4/IL-13-induced M2 macrophage polarization through enhancing STAT6 phosphorylation.

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DATA AVAILABILITY

The data that produced from this study are available from the corresponding author upon reasonable request.

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ACKNOWLEDGEMENTS

We would like to express our special thanks to Dr. Hongyan Wang (Shanghai Institute of Biochemistry and Cell Biology) for her kind assistance to this study.

Funding

This research was supported by the National Natural Science Foundation of China (82000678), the Young Talent Development Plan of Changzhou Health Commission (CZQM2021014), the Changzhou High-Level Medical Talents Training Project (2022CZBJ077), the Foundation of Science and Technology Development of Nanjing Medical University (NMUB2019315), and the Special Fund Project for Laboratory Medicine Research of Jiangsu Medical Association (SYH-3201160–0065).

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  1. Siyuan Zhou and Ting Zhao contributed equally to this work.

Authors and Affiliations

  1. Department of Clinical Laboratory, The Affiliated Changzhou Second People’s Hospital of Nanjing Medical University, Changzhou, 213000, Jiangsu, China

    Siyuan Zhou, Ting Zhao, Xuqiong Chen, Wuwen Zhang, Xiaoyi Zou, Yi Yang, Qinshi Wang, Ping Zhang & Tongbao Feng

  2. Department of Pediatrics, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China

    Tong Zhou

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Contributions

S. Zhou and T. Zhao performed the experiments and prepared the manuscript; T. Feng, T. Zhou, and P. Zhang designed the study and revised the manuscript; S. Zhou, T. Zhao, and W. Zhang analyzed the data; X. Chen, X. Zou, Y. Yang, and Q. Wang helped with the experiments. All authors read and approved the final manuscript.

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Correspondence to Tong Zhou or Tongbao Feng.

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Animal procedures were performed strictly in accordance with the Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Care and Use Committee of Nanjing Medical University.

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Zhou, S., Zhao, T., Chen, X. et al. Runx1 Deficiency Promotes M2 Macrophage Polarization Through Enhancing STAT6 Phosphorylation. Inflammation 46, 2241–2253 (2023). https://doi.org/10.1007/s10753-023-01874-7

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