IFNγ-induced antigen loss in chimeric antigen receptor-T cell therapy

TL;DR

This study reveals how IFNγ, a molecule that usually helps T cells fight cancer, can instead cause cancer cells to hide from treatment in colorectal cancer, and suggests ways to counteract this effect.

Contribution

The study identifies a novel IFNγ-mediated antigen loss mechanism in colorectal cancer cells that limits CAR-T and BiTE therapy efficacy.

Findings

IFNγ secreted by activated CAR-T cells causes bystander colorectal cancer cells to lose GUCY2C antigen.

Antigen loss is mediated through IFNγ receptor, JAK, and cellular stress signaling pathways.

Antigen loss can be reversed using anti-IFNγ antibody, JAK inhibitors, or ER stress relievers.

Abstract

FDA-approved chimeric antigen receptor (CAR)-expressing T cell therapies (CARTs) have revolutionized the treatment of blood cancers. Yet none have been successful for "solid" tumors, such as colorectal cancer (CRC), the 2nd leading cause of cancer deaths. Guanylyl cyclase C (GUCY2C) has emerged as a clinical-stage target for CART and bispecific T-cell engager (BiTE) therapies in CRC. IFNγ has been canonically recognized as beneficial for the effector functions of T cells by enhancing antigen processing and HLA presentation and is essential for CART targeting of solid malignancies by inducing adhesion molecule expression for synapse stabilization.

Using in vitro co-culture systems, conditioned media experiments, cytokine screening, pharmacologic inhibition, CRISPR-Cas9 knockout, and transcriptomic analyses, we investigated mechanisms of GUCY2C antigen loss in CRC cells exposed to activated CART cells.

We identified a novel antigen loss mechanism that limits the efficacy of CART in CRC, in which IFNγ secreted by activated CART cells causes bystander cancer cells to lose GUCY2C. This previously unexplored antigen loss mechanism is mediated through IFNγ receptor, JAK, and cellular stress signaling pathways. This mechanism of antigen loss can be rescued with anti-IFNγ neutralizing antibody, the JAK inhibitor ruxolitinib, or 4-phenylbutyrate (an ER stress reliever).

We revealed a negative effect of IFNγ that uniquely interferes with immunotherapies targeting native surface antigens, such as CART and BiTE therapies, which may be reversed by disrupting stress signaling pathways to enhance solid tumor CART and BiTE immunotherapies.

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