Allogeneic double-negative CAR-T cells inhibit tumor growth without off-tumor toxicities

Abstract

The development of autologous chimeric antigen receptor T (CAR-T) cell therapies has revolutionized cancer treatment. Nevertheless, the delivery of CAR-T cell therapy faces challenges, including high costs, lengthy production times, and manufacturing failures. To overcome this, attempts have been made to develop allogeneic CAR-T cells using donor-derived conventional CD4 + or CD8 + T cells (T convs ), but severe graft-versus-host disease (GvHD) and host immune rejection have made this challenging. CD3 + CD4 − CD8 − double-negative T cells (DNTs) are a rare subset of mature T cells shown to fulfill the requirements of an off-the-shelf cellular therapy, including scalability, cryopreservability, donor-independent anticancer function, resistance to rejection, and no observed off-tumor toxicity including GvHD. To overcome the challenges faced with CAR-T convs , we evaluated the feasibility, safety, and efficacy of using healthy donor–derived allogeneic DNTs as a CAR-T cell therapy platform. We successfully transduced DNTs with a second-generation anti–CD19-CAR (CAR19) without hampering their endogenous characteristics or off-the-shelf properties. CAR19-DNTs induced antigen-specific cytotoxicity against B cell acute lymphoblastic leukemia (B-ALL). In addition, CAR19-DNTs showed effective infiltration and tumor control against lung cancer genetically modified to express CD19 in xenograft models. CAR19-DNT efficacy was comparable with that of CAR19-T convs . However, unlike CAR19-T convs , CAR19-DNTs did not cause alloreactivity or xenogeneic GvHD-related mortality in xenograft models. These studies demonstrate the potential of using allogeneic DNTs as a platform for CAR technology to provide a safe, effective, and patient-accessible CAR-T cell treatment option.

Exosomes in malignant pleural effusion from lung cancer patients impaired the cytotoxicity of double-negative T cells

“…Recently, DNT cells have been successfully applied in lung cancer immunotherapy [5] . Chimeric antigen receptor DNT (CAR-DNT) cells are effective in infiltration into tumor lung cancers [6] . However, DNT cells in lung cancer MPE remain unexplored.…”

Single-cell profiling identifies a spectrum of human unconventional intraepithelial T lineage cells

“…The therapeutic potential of DN T cells as Chimeric antigen receptor (CAR)-T cell is recently explored. It is evidenced that DN CAR-T cells are as effective as conventional CAR-T cells, without inducing toxicity, demonstrating the potential of using allogeneic DN T cells 51 . A relationship between these DN T cells and the uIELs has not yet been conclusively addressed.…”

“…Target cells (W6/32 or OKT3 hybridoma) were labelled with 51 Chromium (Perkin Elmer) for 90 min at 37 °C, washed and added at 10 Table S1. Significantly differentially expressed between the PD-1 + and PD-1population, in both PNT and CB Table S2.…”

CAR-NK Cells: A Chimeric Hope or a Promising Therapy?

“…However, these lead to an increased risk of graft rejection (even failure) or GVHD. Others genetic modifications—such as TCR suppression or the introduction of a suicide gene—are necessary; some researchers even tried using double-negative CAR-T cells (CD4-CD8-) with no rejection or GVHD [ 26 ]. Some clinical studies are underway to test these allogenic CAR-T cells—mostly in B malignancies—with the usual fludarabine–cyclophosphamide (FluCy) lymphodepletion, to which alemtuzumab is added to decrease the risk of immune rejection.…”