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Dryad

Colorectal cancer interleukin-10 blockade scRNA-seq

Abstract

Objective: PD-1 checkpoint inhibition and adoptive cellular therapy have limited success in patients with microsatellite stable colorectal cancer liver metastases (CRLM). We demonstrate that interleukin-10 (IL-10) blockade enhances endogenous T cell and chimeric antigen receptor T (CAR-T) cell anti-tumor function in CRLM slice cultures.

Design: We created organotypic slice cultures from human CRLM (n = 38) and tested the anti-tumor effects of a neutralizing antibody against IL-10 (αIL-10). We evaluated slice cultures with single and multiplex immunohistochemistry, in situ hybridization, single cell RNA sequencing, and time-lapse fluorescent microscopy. In addition, we studied the effects of αIL-10 on carcinoembryonic antigen (CEA)-specific CAR-T cells exogenously administered to both human CRLM slice cultures and a CRLM murine model.

Results: There was little effect of PD-1 blockade in CRLM slice cultures. In contrast, αIL-10 generated 1.8-fold increase in T cell-mediated carcinoma cell death, and increased proportion of CD8+ T cells and inflammatory polarization of macrophages. In addition to effects on endogenous immune cells in human CRLM, αIL-10 also rescued murine CAR-T cell proliferation and cytotoxicity from myeloid cell-mediated immunosuppression. In human CRLM slices, αIL-10 dramatically improved CEA-specific CAR-T cell cytotoxicity, generating nearly 70% carcinoma apoptosis across multiple human tumors. We saw a less dramatic, but similar effect of pretreatment of CAR-T cells with an IL-10 receptor blocking antibody, demonstrating that IL-10 inhibits CAR-T function in the CRLM tumor microenvironment.

Conclusion: Neutralizing the effects of IL-10 in human CRLM has therapeutic potential as a stand-alone treatment and to augment the function of adoptively transferred CAR-T cells.