A Novel Dual CAR-T Cells Specifically Recognize MUC1 and HER2 on Human Pancreatic Cancer with Their Activation

Ning Liu; Qi Luo; Feiyue Xing

doi:10.54097/h62tgg30

Authors

Ning Liu
Qi Luo
Feiyue Xing

DOI:

https://doi.org/10.54097/h62tgg30

Keywords:

Dual CAR-T Cells, Pancreatic Cancer, Specifically Recognize, MUC1 and HER2

Abstract

Aim: The objective of this study was to develop a novel tandem chimeric antigen receptor T (CAR-T) cells targeting both Mucin1 (Muc1) and Human Epidermal Growth Factor Receptor 2 (HER2) on the surface of pancreatic tumor to enhance CAR-T cell specificity and efficacy against this challenging malignancy. Methods: To establish dual-CAR-Jurkat T cells, lentivirus carrying the designed CAR genes was used to infect Jurkat T cells and validated under an inverted fluorescence microscope. Then, in situ immuno-fluorescence staining was utilized to confirm recognition and binding of dual-CAR-Jurkat T cells to Capan-2 cells. Finally, activation of dual-CAR-Jurkat T cells was verified by ELISA after stimulation of Capan-2 cells. Results: The M1H2 CAR-Jurkat T cells targeting Muc1 and HER2 were validated through in situ immuno-fluorescence staining and ELISA. The results showed that compared with the control cells that lack the M1H2 CAR molecules, the many M1H2 CAR-Jurkat T cells adhered tightly to Capan-2 cells, suggesting that the M1H2 CAR-Jurkat cells specifically recognize pancreatic cancer cells. Additionally, the M1H2 CAR-Jurkat cells released higher levels of T cell-activating cytokine IL-2 and cytotoxic cytokine IFN-γ, indicating that the both may maintain the activation and function of the M1H2 CAR-Jurkat T cells. Conclusion: The genetically engineered and modified tandem M1H2 CAR-T cells are successfully established. They can specifically identify and effectively bind pancreatic cancer cells expressing Muc1 and HER2. This might enable men to overcome limitation of current therapeutic approaches and increase therapeutic effects on refractory pancreatic cancer.

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