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Chimeric antigen receptor macrophages activated through TLR4 or IFN-γ receptors suppress breast cancer growth by targeting VEGFR2

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Abstract

Chimeric antigen receptor macrophage (CAR-M) is a promising immunotherapy strategy of anti-tumor due to its high infiltration, direct phagocytosis of tumor cells, immunomodulation of tumor microenvironment (TME) and linkage of innate and adaptive immunity. Here a series of novelly designed CAR-Ms by targeting vascular endothelial growth factor receptor-2 (VEGFR2), which highly expressed in tumor cells and TME, were evaluated. Their activation signals were transduced by Tlr4 or Ifn-γ receptors either alone or in combination, which were designed to mediate M1 polarization of macrophages as the downstream of lipopolysaccharide or Ifn-γ that had been widely reported. Our results showed that VEGFR2-targeting CAR-Ms could be activated under the stimulation of VEGFR2-expressing cells. They exhibited higher expression of CD86, MHCII and TNF-α in vitro and enhanced tumor suppressive abilities in vivo. Implantation of these CAR-Ms into 4T1 breast cancer-bearing mice could obviously inhibit the progression of tumor without significant toxic side effects, especially the group of mmC in which constructed with Tlr4 as the intracellular domain of CAR. In conclusion, this research provides a promising design of CAR that induce macrophages activation by Tlr4 and/or Ifn-γ receptors, and these CAR-Ms could effectively inhibit tumor growth through targeting VEGFR2.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

In the design of CAR-M, Prof Rong Xiang and Yi Shi from Nankai University have given good suggestions. Sincere acknowledgment to them for their kindly support. Elements used for Fig.2b in this article were downloaded from https://smart.servier.com. The Servier Medical Art by Servier is licensed under a Creative Commons Attribution 3.0 Unported License. This work was supported by the Prospective Research Program of the Foundation for the Development of Frontier Technology of Cell Therapy of Changzhou Xitaihu (2022-P-009, Y.L.), the Fundamental Research Funds for the Central Universities (3332022181, Z.D.) and the Bilateral Inter-Governmental S&T Cooperation Project from Ministry of Science and Technology of China (2018YFE0114300, Y.L.).

Funding

This work was supported by the Prospective Research Program of the Foundation for the Development of Frontier Technology of Cell Therapy of Changzhou Xitaihu (2022-P-009, Y.L.), the Fundamental Research Funds for the Central Universities (3332022181, Z.D.) and the Bilateral Inter-Governmental S&T Cooperation Project from Ministry of Science and Technology of China (2018YFE0114300, Y.L.).

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  1. Zhaojun Duan and Zhen Li contributed equally in this work.

Authors and Affiliations

  1. Department of Immunology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, School of Basic Medicine, Peking Union Medical College, Beijing, 100005, China

    Zhaojun Duan, Zhen Li, Ziyuan Wang, Chong Chen & Yunping Luo

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Contributions

The chimeric antigen receptors were designed by ZD and evaluated by ZL and ZD. The conditions for animal experiments were established by ZW. CC helped with the analysis and interpretation of results. The manuscript was written by ZL and edited by ZD and YL. The financial support came from funds of YL and ZD. All authors contributed to manuscript revision, read, and approved the submitted version.

Corresponding author

Correspondence to Yunping Luo.

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Conflict of interest

ZD, ZL and YL have filed a patent on the design and application of the CAR-M cells described in this project for tumor immunotherapy with National Intellectual Property Administration of China. The authors declare no other competing interests.

Ethics approval

All experiments involving animals were approved by the Institute Research Ethics Committee of Peking Union Medical College (2021.9.2/ACVC-A01-2021–039) and carried out under the guidelines on laboratory animals.

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Duan, Z., Li, Z., Wang, Z. et al. Chimeric antigen receptor macrophages activated through TLR4 or IFN-γ receptors suppress breast cancer growth by targeting VEGFR2. Cancer Immunol Immunother 72, 3243–3257 (2023). https://doi.org/10.1007/s00262-023-03490-8

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