Research Of Chimeric Antigen Receptor Gene Modified T Lymphocytes Redirected To Glypican-3 In Hepatocellular Carcinoma Immunotherapy

?Objective?To prepare and develop Glypican 3(GPC3)-targeted CAR modified T lymphocytes and evaluate the anti-tumor effect in vitro and vivo.?Methods?Glypican 3(GPC3)-targeted CAR modified T lymphocytes of healthy and hepatocellular carcinoma patients were developed by lentiviral vectortransduction.The expression of CARs was detected by flow cytometry.The cytotoxic activities of the genetically engineered CAR T cells in vitro were evaluated against various HCC cell lines.The anti-tumor activities of GPC3+ CAR T lymphocytes against GPC3-positive Huh-7 cells were evaluated in the early treatment in vivo.?Results?GPC3 targeted CAR modified T lymphocytes were developed by the lentiviral vector transduction.The positive transduction rates were above 50%.T lymphocytes expressing GPC3 can specifically recognize and kill the hepatocellular carcinoma cells(HCC)cells with positive GPC3,but had no effect on the HCC cell with negative GPC3 in vitro.And during the process of specific killing of tumor cells great amount cytokines including IL-2 and IFN-? were released.T lymphocytes expressing the GPC3-targeted CAR could eradicate HCC xenografts with high level of GPC3 expression in vivo.Furthermore,there was a significant difference between the GPC3+CAR T lymphocytes control and the Mock group.?Conclusions?GPC3-redirected CAR T lymphocytes of both the healthy and hepatocellular carcinoma patients can specifically target and kill GPC3-positive hepatocellular carcinoma cells,which is expected to provide a new and more effective treatment for the clinical treatment of hepatocellular carcinoma.