GPC3-CAR-T Cells for Immunotherapy of Cancer With GPC3 Expression

• Choose appropriate patients with advanced hepatocellular carcinoma,with written consent for this study;
• Perform biopsy to determine the expression of GPC3 of the tumor by western blotting or IHC;
• Collect blood from the patients and isolate mononuclear cells, activate the T cells and transfect the T cells with GPC3/TGFβ targeting CAR (or/and scfv/cytokines-secreting), amplify the number of transfected T cells as needed, test the quality and killing activity of the GPC3/TGFβ-CART cells and then transplant back the patients via systemic or local infusions (via artery or intra-tumor), and follow up closely to collect related results as needed;
• To enhance the killing capability, CD4+ T cells are genetically engineered to express TGFβ-CAR and secret IL7/CCL19 and/or SCFVs against PD1/CTLA4/Tigit; CD8+T cells are constructed to express GPC3-DAP10-CAR with knockdown of PD1/HPK1;
• Evaluate the clinical results as needed.
• Will also perform the similar clinical trial on lung squamous carcinoma with the GPC3 expression.

Inclusion criteria

• patients with advanced HCC,which express GPC3 protein.
• Life expectancy >12 weeks
• Child-Pugh-Turcotte score <7
• Adequate heart,lung,liver,kidney function
• Available autologous transduced T cells with greater than or equal to 20% expression of GPC3 CAR determined by flow-cytometry and killing of GPC3-positive targets greater than or equal to 20% in cytotoxicity assay
• Informed consent explained to, understood by and signed by patient/guardian. Patient/guardian given copy of informed consent. -

Exclusion criteria

• Had accepted gene therapy before;
• Tumor size more than 25cm;
• Severe virus infection such as HBV,HCV,HIV,et al
• Known HIV positivity
• History of liver transplantation
• Active infectious disease related to bacteria, virus,fungi,et al
• Other severe diseases that the investigators consider not appropriate;
• Pregnant or lactating women
• Systemic steroid treatment (greater than or equal to 0.5 mg prednisone equivalent/kg/day)
• Other conditions that the investigators consider not appropriate. -