Construction and Characterization of Cadherin 6 (CDH6)-Targeting Chimeric Antigen Receptor (CAR) Modified T Cells.

Abstract

In this study, we constructed cadherin 6 (CDH6)-targeting chimeric antigen receptor (CAR) modified T cells (CAR-T cells) and investigated their target-specific recognition and tumor-specific cytocidal effect through in vitro approach. CDH6 expression at the transcriptional level and its correlation with clinicopathological parameters in various tumor types were analyzed using online bioinformatics tool. Conventional molecular cloning method was used to construct the lentiviral vector encoding CDH6-specific CAR and the lentivirus was prepared using 3-plasmid transient cotransfection method. CDH6-targeting CAR-T cells were prepared using centrifugal infection method, and the specific recognition and cytocidal effects of CAR-T cell targets were investigated through in vitro co-culture experiments. At the transcription level, CDH6 was significantly overexpressed in ovarian cancer tissues (P < 0.05). Although it was not correlated with tumor stage and patient's prognosis, the overexpression of CDH6 was positively associated with the expression of paired-box 8 (PAX8), a lineage-specific transcription factor. In the present study, we successfully established CDH6-targeting CAR-T cells that can secrete effector cytokines and produce specific cytocidal effects after being co-cultured with CDH6-positive ovarian cancer cells in vitro. Thus, CDH6, as a lineage-specific factor of ovarian tissue, may be an ideal target for CAR-T cell therapy of ovarian cancer.