Abstract
Chimeric antigen receptor-modified T cell (CAR-T) therapy is a newly developed adoptive treatment for cancer in recent years that has shown potential for curative therapeutic efficacy in patients with hematological malignancies. However, the efficacy against solid tumors was very limited, the reasons are complex and have not been fully understood. In this study, CAR-T cells with 4-1BB or CD28 co-stimulatory molecules targeting carbonic anhydrase IX (CAIX) in the presence of 0, 10, 50, and 100 µM Docosahexaenoic acid (C22:6, DHA) has been used to treat renal cancer cells to illustrate the anti-tumor efficacy to solid tumor cells. The cytotoxicity of CAIX-CAR-T cells against cancer cells in the presence of DHA was determined by real-time cell assay (RTCA). Cytokines released by CAR-T cells were analyzed by Enzyme linked immunosorbent assay (ELISA). CAR-T cell apoptosis and exhaustion induced by DHA were determined by flow cytometry. Furthermore, signaling molecules involved in cell apoptosis and exhaustion were examined by Western blot analysis. This study showed that both 4-1BB and CD28-derived CAIX-CAR-T cells displayed efficient anti-tumor ability against renal carcinoma OSRC-2 cells. DHA attenuated cytotoxicity of CAR-T cells by inducing cell apoptosis and exhaustion in an opposite regulatory manner of AKT/PKB (protein kinase B) activation, which may provide a strategy to ameliorate CAR-T efficacy against solid tumors.
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Acknowledgements
The authors thank Qianqian Dai, Cancer Institute, Xuzhou Medical University, for helping with flow cytometry operation, and data acquisition.
Funding
This work was supported by the National Natural Science Foundation of China [81773253, 81972242]; the Jiangsu Province Social Development Projects [BE2020641]; the Natural Science Foundation of Jiangsu Province [BK20211057]; the Natural Science Project of Jiangsu Provincial Education Department [19KJB310018]; the Research Foundation of Xuzhou Medical University [D2019023]; the Xuzhou Science and Technology Bureau projects [KC19058]; Youth Technology Innovation Team of Xuzhou Medical University [TD202003]; the Project of Invigorating Health Care through Science, Technology and Education [ZDXKA2016014]; National Demonstration Center for Experimental Basic Medical Science Education (Xuzhou Medical University) [202010313049Y].
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Conceptualization, SS.S., and JN.Z.; methodology, XG.G. and Q.Z.; software, XG.G.; validation, YF.W., YX.Z. and YZ.Z.; formal analysis, Q.Z.; investigation, YF.W., YX.Z., YZ.Z., H.X., WX.Z. and C.H., BX.W.; resources, YF.W., SS.S., JN.Z. and Q.Z.; data curation, YF.W.; writing—original draft preparation, YF.W.; writing—review and editing, YF.W., and SS.S.; visualization, JN.Z.; supervision, JN.Z. and SS.S.; project administration, JN.Z. and Q.Z.; funding acquisition, SS.S., YF.W., JN.Z. and Q.Z. All authors have read and agreed to the published version of the manuscript.
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Wang, Y., Zhao, Y., Zhu, Y. et al. DHA attenuates CAR-T cell efficacy through aggravating exhaustion and inducing apoptosis. Biologia 78, 1141–1152 (2023). https://doi.org/10.1007/s11756-023-01326-5
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DOI: https://doi.org/10.1007/s11756-023-01326-5