Crucially, functional assays demonstrated that NKG2D-CAR T cell phenotype, activity, and cytokine secretion remained unaffected and its activity against three-dimensional OS-spheroids was preserved under in vitro hypoxic conditions. Although these findings challenge the idea that hypoxia alone compromises in vitro NKG2D-CAR T efficacy in OS, further studies are needed on how hypoxia interacts with multiple factors of the TME that could modulate CAR T cell behavior.

CYAD-02 presented an higher engraftment and an improved clinical activity (17% objective response rate) compared to CYAD-01 (no objective response). Altogether, our data provide proof of principle that knock-down of MICA/B can enhance CAR T-cell persistence and efficacy while maintaining a good safety profile.

LIN28 overexpression maintained human T cell phenotype markers and functionality but impaired the antitumoral cytotoxicity of NKG2D-CAR T cells both in vitro and in vivo. These findings highlight the intricate relationship between LIN28/let-7 axis and human T cell functionality, including in CAR T cell therapy.

The results of immunohistochemistry and immunofluorescence assays showed that the combined HA-Mn-CAP Gel and NKG2D/CAR-T significantly increased the proliferation and infiltration of T cells, especially for CD8+ cells. Therefore, the new promising strategy of increasing the target ligand expression and adjusting TME before administering CAR-T cells is suitable for treating solid tumors.

P=N/A, N=9, Recruiting, Cancer Institute and Hospital, Chinese Academy of Medical Sciences

P1, N=18, Recruiting, Antonio Pérez Martínez | Not yet recruiting --> Recruiting

P1, N=1, Terminated, Celyad Oncology SA | Unknown status --> Terminated; Difficulties in patient recruitment and clinical strategy modification based on new clinical data generated in the NKR-2 early development.

P1, N=12, Not yet recruiting, Peking University

In sum, our study reveals the role of DKK1 in remodeling GC TIME and regulating the expression levels of NKG2DLs in GC. We also provide a promising treatment strategy of combining DKK1 inhibition with NKG2D-CAR-T cell therapy, which could bring new breakthroughs for GC immunotherapy.

In vitro, T cells transduced with these constructs displayed cytotoxic activity, proliferated, and secreted cytokines in presence of WT and CD19 KO B-ALL cancer cells. Since NKG2DL are expressed in many cancer indications and absent from healthy tissues, use of NKG2DL-based dual CAR T cells is an attractive approach to improve anti-tumor efficacy in solid indications.

Treatment with a multiple CYAD-01 infusion schedule without preconditioning is well tolerated and shows anti-leukaemic activity, although without durability outside of patients bridged to allogeneic HSCT. These phase 1 data support the proof-of-concept of targeting NKG2D ligands by CAR T-cell therapy. Further clinical studies with NKG2D-based CAR T-cells are warranted, potentially via combinatorial antigen targeted approaches, to improve anti-tumour activity.

P1, N=18, Recruiting, jianming xu