LAUSR

Simple Summary One promising avenue for the treatment of colorectal cancer is the reinvigoration of pre-existing anti-tumor T-cell responses by overcoming inhibitions that arise from the T-cell expression of immune checkpoints. This study aims to perform an in-depth assessment of human colorectal cancer, the immune checkpoint landscape in tumor-infiltrating T cells and their respective ligands in tumor cells and myeloid cells; this is achieved using multiparametric flow cytometry on dissociated fresh tumors from 40 patients and in situ immunohistochemistry. Our results highlight the strong expression of TIGIT and its ligand, CD155, in tumors, suggesting the role of the TIGIT/CD155 axis in colorectal cancer. Moreover, unsupervised clustering allowed us to identify two distinct subgroups of colorectal cancer patients according to the co-expression of several immune checkpoints on T-lymphocyte subpopulations. Altogether, our findings support the development of colorectal cancer immunotherapies targeting TIGIT, which could be used in combination immune checkpoint therapy in colorectal cancer. Abstract While immune checkpoint (IC) therapies, particularly those targeting the PD-1/PD-L1 axis, have revolutionized the treatment of melanoma and several other cancers, their effect remains very limited in colorectal cancer (CRC). To define a comprehensive landscape of ICs in the human CRC tumor microenvironment (TME), we evaluated, using multiparametric flow cytometry, their ex vivo expression via tumor-infiltrating lymphocytes (TILs) (n = 40 CRCs) as well as that of their respective ligands on tumor and myeloid cells (n = 29). Supervised flow cytometry analyses showed that (i) most CD3+ TILs expressed PD-1 and TIGIT and, to a lesser extent, Tim-3, Lag3 and NKG2A, and (ii) EpCAM+ tumor cells and CD11b+ myeloid cells differed in their IC ligand expression profile, with a strikingly high expression of CD155 by tumor cells. An in situ analysis of IC and their ligands using immunohistochemistry on paraffin sections of CRC confirmed the overexpression of TIGIT and its ligand, CD155, in the TME. Most interestingly, an unsupervised clustering analysis of IC co-expression on CD4+ and CD8+ TILs identified two tumor subgroups, named IChigh and IClow. Altogether, our findings highlight the TIGIT/CD155 axis as a potential target that could be used in combination IC therapy in CRC.