Chemokine Receptors CCR6 and PD1 Blocking scFv E27 Enhances Anti-EGFR CAR-T Therapeutic Efficacy in a Preclinical Model of Human Non-Small Cell Lung Carcinoma

• Published in: International journal of molecular sciences 2023-03, Vol.24 (6), p.5424
• Main Authors: Wang, Jing, Wang, Yanan, Pan, Hanyu, Zhao, Lin, Yang, Xinyi, Liang, Zhiming, Shen, Xiaoting, Zhang, Jing, Yang, Jinlong, Zhu, Yuqi, Xun, Jingna, Liang, Yue, Lin, Qinru, Liang, Huitong, Li, Min, Zhu, Huanzhang
• Format: Article
• Language: English
• Subjects: Animals; Antigens; Apoptosis; Breast cancer; Cancer; Cancer therapies; CAR T cell therapy; Carcinoma; Carcinoma, Non-Small-Cell Lung - pathology; CCR6 protein; Cell adhesion & migration; Cell growth; Cell Line, Tumor; checkpoint; chemokine; Chemokine receptors; Chemokines; Chimeric antigen receptors; Clinical trials; Cytotoxicity; EGFR; Epidermal growth factor; Epidermal growth factor receptors; ErbB Receptors - metabolism; Genotype & phenotype; Growth factors; Health aspects; Humans; Immunodeficiency; Immunotherapy; Immunotherapy, Adoptive - methods; Inflammation; Interleukin 2; Kinases; Lung cancer; Lung cancer, Non-small cell; Lung carcinoma; Lung Neoplasms - pathology; Lungs; Lymphocytes; Lymphocytes T; Medical prognosis; Metastases; Mice; Mice, Inbred NOD; migration; Non-small cell lung carcinoma; NSCLC; PD-1 protein; Pharmacology; Programmed Cell Death 1 Receptor - metabolism; Receptors, CCR6; Receptors, Chemokine; Receptors, Chimeric Antigen; Solid tumors; T cells; Tumor cells; Tumor necrosis factor-α; Tumors; Xenograft Model Antitumor Assays; Xenografts; Xenotransplantation; γ-Interferon
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms24065424

Chimeric antigen receptor (CAR)-T cells, a therapeutic agent for solid tumors, are not completely effective due to a lack of infiltration of T cells into the tumor site and immunity caused by Programmed Death Receptor 1(PD1). Here, an epidermal growth factor receptor (EGFR) CAR-T cell was engineered to express the chemokine receptor CCR6 and secrete PD1 blocking Single-chain antibody fragment (scFv) E27 to enhance their anti-tumor effects. The findings showed that CCR6 enhanced the migration of EGFR CAR-E27-CCR6 T cells in vitro by the Transwell migration assay. When incubated with tumor cells, EGFR CAR-E27-CCR6 T cells specifically exerted potent cytotoxicity and produced high levels of pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), and interferon-γ (IFN-γ). A non-small cell lung carcinoma (NSCLC) cell line-derived xenograft model was constructed by implanting modified A549 cell lines into immunodeficient NOD. (NSG) mice. In comparison with traditional EGFR CAR-T cells, live imaging indicated that EGFR CAR-E27-CCR6 T cells displayed superior anti-tumor function. In addition, the histopathological examination of mouse organs showed no obvious organic damage. Our findings confirmed that PD1 blocking and CCR6 can enhance the anti-tumor function of EGFR CAR-T cells in an NSCLC xenograft model, providing an effective treatment strategy to improve the efficacy of CAR-T in NSCLC.