通用型CD19 CAR-T的体外构建及初步功能鉴定

In Vitro Construction and Preliminary Functional Identification of Universal CD19 CAR-T

CAR-T免疫细胞治疗已经在血液肿瘤领域取得突破性进展。然而,目前上市和国内临床试验的CAR-T细胞均来自肿瘤患者自身,即自体型CAR-T。因受制于患者T细胞的质量和数量、制备周期长且价格昂贵等原因,很难将其进行大规模临床应用。该研究利用CRISPR/Cas9基因编辑技术敲除健康人脐带血来源T细胞的TCR分子和HLA-I类分子,避免异基因细胞治疗引起的免疫排斥,通过慢病毒载体转导CAR基因,制备通用型CD19 CAR-T细胞药物。体外验证可减少免疫排斥,并在体外证明有较强的靶细胞杀伤作用。该方法可提供一种制备通用型CAR-T细胞的途径,有望使更多患者得到及时治疗,使重复治疗成为可能;另外,它降低了CAR-T疗法的制造成本,从而减轻了患者的治疗负担;最后,它可为临床治疗提供实验依据。

CAR-T immune cell therapy has made breakthrough progress in the field of hematological tumors.However,the CAR-T cells currently on the market and in domestic clinical trials are all derived from tumor patients themselves,that is,autologous CAR-T.Restricted by the quality and quantity of patient T cells,long preparation cycle and high price,it is difficult to apply them in large-scale clinical applications.In this study,CRISPR/Cas9 gene editing technology was used to knock out TCR molecules and HLA-I molecules in T cells derived from healthy human umbilical cord blood to avoid immune rejection caused by allogeneic cell therapy.CAR gene was transduced by lentiviral vector to prepare universal CD19 CAR-T cell drugs.It is proved that it can reduce immune rejection in vitro and has a strong killing effect on target cells in vitro.This method provides a way to produce universal CAR-T cells.Firstly,it is expected to make more patients to receive timely treatment and makes it possible to repeat treatment.Secondly,it reduces the manufacturing cost of CAR-T therapy,thereby reducing the burden of patients.Finally,it provides experimental basis for clinical treatment.