BiTE-Secreting CAR-γδT as a Dual Targeting Strategy for the Treatment of Solid Tumors

HLA-G is considered as an immune checkpoint protein and a tumor-associated antigen. In the previous work, it is reported that CAR-NK targeting of HLA-G can be used to treat certain solid tumors. However, the frequent co-expression of PD-L1 and HLA-G) and up-regulation of PD-L1 after adoptive immunotherapy may decrease the effectiveness of HLA-G-CAR. Therefore, simultaneous targeting of HLA-G and PD-L1 by multi-specific CAR could represent an appropriate solution. Furthermore, gamma-delta T (gamma delta T) cells exhibit MHC-independent cytotoxicity against tumor cells and possess allogeneic potential. The utilization of nanobodies offers flexibility for CAR engineering and the ability to recognize novel epitopes. In this study, V delta 2 gamma delta T cells are used as effector cells and electroporated with an mRNA-driven, nanobody-based HLA-G-CAR with a secreted PD-L1/CD3 epsilon Bispecific T-cell engager (BiTE) construct (Nb-CAR.BiTE). Both in vivo and in vitro experiments reveal that the Nb-CAR.BiTE-gamma delta T cells could effectively eliminate PD-L1 and/or HLA-G-positive solid tumors. The secreted PD-L1/CD3 epsilon Nb-BiTE can not only redirect Nb-CAR-gamma delta T but also recruit un-transduced bystander T cells against tumor cells expressing PD-L1, thereby enhancing the activity of Nb-CAR-gamma delta T therapy. Furthermore, evidence is provided that Nb-CAR.BiTE redirectes gamma delta T into tumor-implanted tissues and that the secreted Nb-BiTE is restricted to the tumor site without apparent toxicity.

Automated summary

HLA-G is an immune checkpoint protein and a tumor-associated antigen. Previous research has shown the potential of using CAR-NK targeting HLA-G to treat certain solid tumors. However, co-expression of PD-L1 and HLA-G and up-regulation of PD-L1 after immunotherapy may reduce the effectiveness of HLA-G-CAR. Therefore, a multi-specific CAR targeting both HLA-G and PD-L1 could provide a solution.