A Vδ1+γδ T cell therapy (DOT) effectively lysed CRC lines and organoids, and exhibited efficacy in a xenograft CRC model. Infiltrating DOTs showed reductions in activating NK receptors (NKG2D, DNAM1) and cytotoxic markers, with increases in TIGIT+PD-1+ cells. These features were reflected in scRNAseq of endogenous tumor-infiltrating γδ T cells in patient samples. Co-exposure to TIGIT and PD-1 ligands decreased cytotoxicity, and dual ICB strikingly improved the efficacy of DOT in vivo. DOT cytotoxicity was partially dependent on NKG2D, and enhancing NKG2DL expression on tumor cells via butyrate treatment improved cell killing.
Contributed by Morgan Janes
ABSTRACT: Colorectal cancer (CRC) remains a challenge for current immunotherapies. Vδ1+ γδ T cells offer a promising alternative because of their HLA-I-independent cytotoxicity and natural tissue tropism. We developed Delta One T (DOT) cells, a Vδ1+ γδ T cell-based adoptive cell therapy clinically explored for hematological malignancies but not yet for solid tumors. Here we demonstrate the capacity of DOT cells to target CRC cell lines and patient-derived specimens and organoids in vitro and to control tumor growth in an orthotopic xenograft model of CRC. Notwithstanding, we found tumor-infiltrating DOT cells to exhibit a dysregulated balance of cytotoxic and inhibitory receptors that paralleled that of endogenous Vδ1+ tumor-infiltrating lymphocytes and limited their cytotoxicity. To maximize efficacy, we unveil two strategies, increasing targeting through upregulation of NKG2D ligands upon butyrate administration and blocking the checkpoints TIGIT and PD1, which synergistically unleashed DOT cell cytotoxicity. These findings support DOT cell-based combinatorial approaches for CRC treatment.
Author Info: (1) Gulbenkian Institute for Molecular Medicine, Lisbon, Portugal. rafael.blanco@gimm.pt. (2) Gulbenkian Institute for Molecular Medicine, Lisbon, Portugal. (3) Gulbenkian Institut
Author Info: (1) Gulbenkian Institute for Molecular Medicine, Lisbon, Portugal. rafael.blanco@gimm.pt. (2) Gulbenkian Institute for Molecular Medicine, Lisbon, Portugal. (3) Gulbenkian Institute for Molecular Medicine, Lisbon, Portugal. (4) Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands. (5) Gulbenkian Institute for Molecular Medicine, Lisbon, Portugal. (6) University of Bordeaux, CNRS, ImmunoConcEpT, UMR 5164, Bordeaux, France. (7) Hospital de Santa Maria, Centro Hospitalar Lisboa Norte, Lisbon, Portugal. (8) Hospital de Santa Maria, Centro Hospitalar Lisboa Norte, Lisbon, Portugal. (9) Hospital de Santa Maria, Centro Hospitalar Lisboa Norte, Lisbon, Portugal. (10) University of Bordeaux, CNRS, ImmunoConcEpT, UMR 5164, Bordeaux, France. Equipe labelise LIGUE Contre le Cancer, Bordeaux, France. (11) Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands. (12) Gulbenkian Institute for Molecular Medicine, Lisbon, Portugal. (13) Gulbenkian Institute for Molecular Medicine, Lisbon, Portugal. bssantos@medicina.ulisboa.pt. Faculdade de Medicina da Universidade de Lisboa, Lisbon, Portugal. bssantos@medicina.ulisboa.pt.
