The Cytotoxicity Of Anti-EGFR Chimeric Antigen Receptor T Cells For Glioma Cell Lines

Chimeric antigen receptor(CAR)T cell therapy is a kind of adoptive cellular immunotherapy which use genetically modified autologous T cells to kill their own tumor cells specifically.The exogenous protein gives T cells a new active function:to identify tumor associated antigen on the surface specifically and active T cells directly,without major histocompatibility complex(MHC)and stimulating molecules.Glioma is the most common and aggressive primary intracranial tumor in adults.Its pathogenesis is unclear.Patients treated with traditional surgical resection,combined postoperative chemotherapy and radiotherapy are likely to recurrence after a few months,once again,the probability of recurrence after surgery is still high.It is reported that high expression of epithelial growth factor receptor(EGFR)has a close relationship with the occurrence and development of glioma,40-50%of brain glioma have a overexpression of EGFR.Although the antineoplastic drugs which are targeted EGFR and its signaling pathway have made certain progress in colon cancer and other cancers,they are failed in treating glioma.On the other hand,the low expression of EGFR provides chimeric antigen receptor T cell therapy a good target.And brain drug delivery can take advantage of the blood brain barrier to prevent T cells to spread to other parts of the body,cause damage of tissues which are express EGFR,too.Thereore,study focuses on the anti-EGFR CAR-T cells for local treatment of glioma is very meaningful.This study designed four sequences of the second generation anti-EGFR CAR,and verify their expression in 293 T cells.This way,we selected the two best structures.After optimizing the condition of lentivirus packaging,infection,lentivirus vectors and concentration,we modified primary human T lymphocytes by the two sequences above.And finally get a transfection rate of almost 20%.In the end,in vitro cytotoxicity-tests were carried out among cell lines which have a high or lower lever expression of EGFR.The results demonstrated that our constracted CAR-T effectively killed the glioma cell lines with high EGFR expression.This study paves a fundation of local treatment of specific lesion targeting EGFR in the future.