Inhibition of VEGF signaling prevents exhaustion and enhances anti-leukemia efficacy of CAR-T cells via Wnt/β-catenin pathway
Abstract Background Current challenges in Chimeric Antigen Receptor (CAR) -T cell therapy for hematological cancers include T cell exhaustion and limited persistence, which contribute to cancer relapse. Methods The effects of Axitinib, a VEGFR inhibitor, on the biological functions of CAR-T cells in...
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BMC
2025-04-01
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| Series: | Journal of Translational Medicine |
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| Online Access: | https://doi.org/10.1186/s12967-024-05907-z |
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| author | Suwan Wu Yiyi Wei Yingqi Qiu Kexin Ai Mu Chen Hao Wang Honghao Zhang Qingyan Cen Peiyun Liao Xiangyang Ding Xiaoling Xie Yuhua Li |
| author_facet | Suwan Wu Yiyi Wei Yingqi Qiu Kexin Ai Mu Chen Hao Wang Honghao Zhang Qingyan Cen Peiyun Liao Xiangyang Ding Xiaoling Xie Yuhua Li |
| author_sort | Suwan Wu |
| collection | DOAJ |
| description | Abstract Background Current challenges in Chimeric Antigen Receptor (CAR) -T cell therapy for hematological cancers include T cell exhaustion and limited persistence, which contribute to cancer relapse. Methods The effects of Axitinib, a VEGFR inhibitor, on the biological functions of CAR-T cells in vitro and in vivo were investigated by comparing CAR-T cells pre-treated ex vivo with Axitinib, as well as utilizing a B-ALL mouse model. Real-time quantitative PCR and Western blotting were employed to detect the expression of molecules related to differentiation, exhaustion, and the Wnt pathway in CAR-T cells. Flow cytometry was used to assess changes in CAR-T cell differentiation, exhaustion, activation, apoptosis, proliferation, and cytokine secretion. Western blotting and flow cytometry were used to assess changes in VEGFR expression. Bioluminescence imaging, flow cytometry, and immunohistochemistry (IHC) analysis were used to evaluate changes in tumor burden in mice receiving different treatments, while hematoxylin and eosin (H&E) staining were used to monitor histological changes in the liver and spleen of mice. Results Axitinib treatment notably reduced CAR-T cell exhaustion and terminal differentiation both under tonic signaling and tumor antigen exposure scenarios. Furthermore, CAR-T cells pretreated with Axitinib demonstrated enhanced anti-tumor efficacy and prolonged survival in vivo. Mechanistically, Axitinib treatment upregulated the Wnt/β-catenin signaling pathway in CAR-T cells. Using agonists/inhibitors of the Wnt/β-catenin pathway could respectively mimic or counteract the effects of Axitinib on CAR-T cell exhaustion and differentiation. CAR-T cells treated with Axitinib can inhibit the VEGFR2 pathway. CAR-T cells treated with anti-VEGFR2 antibody can activate the Wnt/β-catenin pathway and prevent CAR-T cell exhaustion. Conclusion Axitinib confers resistance to exhaustion in CAR-T cells by modulating the Wnt/β-catenin signaling pathway. |
| format | Article |
| id | doaj-art-b6b9b8ad157e4140b7968c18d18c4607 |
| institution | Kabale University |
| issn | 1479-5876 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj-art-b6b9b8ad157e4140b7968c18d18c46072025-08-20T03:52:19ZengBMCJournal of Translational Medicine1479-58762025-04-0123111710.1186/s12967-024-05907-zInhibition of VEGF signaling prevents exhaustion and enhances anti-leukemia efficacy of CAR-T cells via Wnt/β-catenin pathwaySuwan Wu0Yiyi Wei1Yingqi Qiu2Kexin Ai3Mu Chen4Hao Wang5Honghao Zhang6Qingyan Cen7Peiyun Liao8Xiangyang Ding9Xiaoling Xie10Yuhua Li11Department of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityDepartment of Hematology, Zhujiang Hospital, Southern Medical UniversityAbstract Background Current challenges in Chimeric Antigen Receptor (CAR) -T cell therapy for hematological cancers include T cell exhaustion and limited persistence, which contribute to cancer relapse. Methods The effects of Axitinib, a VEGFR inhibitor, on the biological functions of CAR-T cells in vitro and in vivo were investigated by comparing CAR-T cells pre-treated ex vivo with Axitinib, as well as utilizing a B-ALL mouse model. Real-time quantitative PCR and Western blotting were employed to detect the expression of molecules related to differentiation, exhaustion, and the Wnt pathway in CAR-T cells. Flow cytometry was used to assess changes in CAR-T cell differentiation, exhaustion, activation, apoptosis, proliferation, and cytokine secretion. Western blotting and flow cytometry were used to assess changes in VEGFR expression. Bioluminescence imaging, flow cytometry, and immunohistochemistry (IHC) analysis were used to evaluate changes in tumor burden in mice receiving different treatments, while hematoxylin and eosin (H&E) staining were used to monitor histological changes in the liver and spleen of mice. Results Axitinib treatment notably reduced CAR-T cell exhaustion and terminal differentiation both under tonic signaling and tumor antigen exposure scenarios. Furthermore, CAR-T cells pretreated with Axitinib demonstrated enhanced anti-tumor efficacy and prolonged survival in vivo. Mechanistically, Axitinib treatment upregulated the Wnt/β-catenin signaling pathway in CAR-T cells. Using agonists/inhibitors of the Wnt/β-catenin pathway could respectively mimic or counteract the effects of Axitinib on CAR-T cell exhaustion and differentiation. CAR-T cells treated with Axitinib can inhibit the VEGFR2 pathway. CAR-T cells treated with anti-VEGFR2 antibody can activate the Wnt/β-catenin pathway and prevent CAR-T cell exhaustion. Conclusion Axitinib confers resistance to exhaustion in CAR-T cells by modulating the Wnt/β-catenin signaling pathway.https://doi.org/10.1186/s12967-024-05907-zChimeric antigen receptor T cellsAcute lymphoblastic leukemiaVascular endothelial growth factor receptorDifferentiationExhaustion |
| spellingShingle | Suwan Wu Yiyi Wei Yingqi Qiu Kexin Ai Mu Chen Hao Wang Honghao Zhang Qingyan Cen Peiyun Liao Xiangyang Ding Xiaoling Xie Yuhua Li Inhibition of VEGF signaling prevents exhaustion and enhances anti-leukemia efficacy of CAR-T cells via Wnt/β-catenin pathway Journal of Translational Medicine Chimeric antigen receptor T cells Acute lymphoblastic leukemia Vascular endothelial growth factor receptor Differentiation Exhaustion |
| title | Inhibition of VEGF signaling prevents exhaustion and enhances anti-leukemia efficacy of CAR-T cells via Wnt/β-catenin pathway |
| title_full | Inhibition of VEGF signaling prevents exhaustion and enhances anti-leukemia efficacy of CAR-T cells via Wnt/β-catenin pathway |
| title_fullStr | Inhibition of VEGF signaling prevents exhaustion and enhances anti-leukemia efficacy of CAR-T cells via Wnt/β-catenin pathway |
| title_full_unstemmed | Inhibition of VEGF signaling prevents exhaustion and enhances anti-leukemia efficacy of CAR-T cells via Wnt/β-catenin pathway |
| title_short | Inhibition of VEGF signaling prevents exhaustion and enhances anti-leukemia efficacy of CAR-T cells via Wnt/β-catenin pathway |
| title_sort | inhibition of vegf signaling prevents exhaustion and enhances anti leukemia efficacy of car t cells via wnt β catenin pathway |
| topic | Chimeric antigen receptor T cells Acute lymphoblastic leukemia Vascular endothelial growth factor receptor Differentiation Exhaustion |
| url | https://doi.org/10.1186/s12967-024-05907-z |
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