Clinical Grade Manufacture of CYAD-101, a NKG2D-based, First in Class, Non–Gene-edited Allogeneic CAR T-Cell Therapy

Abstract

Allogeneic chimeric antigen receptor (CAR) T holds the promise of taking this therapeutic approach to broader patient populations while avoiding the intensive manufacturing demands of autologous cell products. One limitation to delivering an allogeneic CAR T is T-cell receptor (TCR) driven toxicity. In this work, the expression of a peptide to interfere with TCR signaling was assessed for the generation of allogeneic CAR T cells. The expression of a truncated CD3ζ peptide was shown to incorporate into the TCR complex and to result in blunted TCR responses. When coexpressed with a natural killer group 2D (NKG2D) CAR, the allogeneic T cells (called CYAD-101) failed to induce graft-versus-host disease in mouse models while maintaining antitumor activity driven by the CAR in vitro and in vivo. Two clinical grade discrete batches of CYAD-101 cells were produced of single donor apheresis resulting in 48 billion CAR T cells sufficient for the entire dose-escalation phase of the proposed clinical trial. The 2 batches showed high consistency producing a predominantly CD4 + T-cell population that displayed an effector/central memory phenotype with no evidence of exhaustion markers expression. These clinical grade CYAD-101 cells secreted cytokines and chemokines in response to ligands expressing target cells in vitro, demonstrating effector function through the CAR. Moreover, CYAD-101 cells failed to respond to TCR stimulation, indicating a lack of allogeneic potential. This bank of clinical grade, non-gene-edited, allogeneic CYAD-101 cells are used in the alloSHRINK clinical trial (NCT03692429).

NKG2D Fine-Tunes the Local Inflammatory Response in Colorectal Cancer

“…NKG2D ligands can stimulate adaptive and innate immune responses by engaging NKG2D on αβ T cells, γδ T cells, iNKT cells, and innate lymphoid cells (ILCs), including natural killer (NK) cells. Targeting NKG2D in CAR cell therapies employing NK cells, γδ T cells or αβ T cells has proved to be a viable approach by several preclinical studies [ 12 , 13 , 14 ] and is currently in its early clinical phase (NCT04623944, NCT05247957), including in patients with refractory metastatic CRC (NCT05213195, NCT05248048) [ 15 , 16 , 17 , 18 , 19 ]. While encouraging anti-tumor activity was recorded against hematological malignancies [ 20 ], limited data are available in the case of solid tumor and metastatic CRC in particular [ 21 ].…”

Metabolic profiling reveals altered tryptophan metabolism in patients with kawasaki disease

“…On the other hand, in the kynurenine pathway, IDO1 is the primary rate-limiting enzyme. Furthermore, several recent studies have demonstrated that the expression of IDO1 is elevated in response to inflammatory stimuli, such as type I and II interferons ( Puccetti and Grohmann, 2007 ), prostaglandins ( Jones et al, 2015 ), or microbial stimuli, such as lipopolysaccharides ( Michaux et al, 2022 ). Meanwhile, IDO1 is regarded as a target gene to regulate overactive immune responses in human autoimmune diseases ( Pan et al, 2008 ; Platten et al, 2012 ; Kasper et al, 2016 ).…”

Allosteric Inhibition of the T Cell Receptor by a Designed Membrane Ligand

“…PITCR could also be used to overcome a critical limitation of allogeneic CAR T therapy, since TCR inhibition is required to prevent graft-versus-host disease resulting from side effects of this therapy. The use of PITCR would additionally overcome the risk resulting from genetic manipulation (i.e., viral transfection) of allogeneic T cells before injection into patients (Michaux et al, 2022).…”