关键词: 间皮素, 人源化scFv, CAR-T, 实体瘤

Abstract: Despite that CAR-T cell (chimeric antigen receptor-modified T cell) therapy has achieved remarkable outcomes in the treatment of hematological malignancies, there are still lots of obstructions in the treatment of solid tumors. Mesothelin is a tumor associated antigen over-expressing on tumor cell surface such as mesothelioma, ovarian, pancreatic. Most of the current studies about CAR-T cells targeting Mesothelin use scFv (single-chain variable fragment) of mouse are to build expression vector by gene editing, which shows some shortcomings in simulating clinical treatment and obstacles of translation of research to clinical treatment, so it is very necessary to develop humanized study. We constructed a CAR vector with a humanized anti-mesothelin scFv called P4, packaged the lenti-virus and used them to transfect human peripheral blood CD3+ T cells. We successfully produced P4 CAR-T cells with high and steady transfection efficiency. PCR results indicated that Mesothelin was expressed in human cervical cancer tumor cell line HeLa. In vitro experiments demonstrated that our P4 CAR-T cells could effectively kill HeLa cells. In vivo experiments showed that infusion of P4 CAR-T cells slowed the growth of established tumors slightly and prolonged the survival of tumor-bearing mice in comparison to the control group. Here we reported that the P4 CAR-T cell can be used as a potential therapeutic strategy against Mesothelin positive tumor, and it provided a basis for further research on synergistic strategies to improve the therapeutic effect in vivo and clinical treatment.

Key words: Mesothelin, humanized scFv, CAR-T, solid tumor