EZH2 Inhibition in Ewing Sarcoma Upregulates GD2 Expression for Targeting with Gene-Modified T Cells

摘要

Chimeric antigen receptor (CAR) engineering of T cells allows one to specifically target tumor cells via cell surface antigens. A candidate target in Ewing sarcoma is the ganglioside G , but heterogeneic expression limits its value. Here we report that pharmacological inhibition of Enhancer of Zeste Homolog 2 (EZH2) at doses reducing H3K27 trimethylation, but not cell viability, selectively and reversibly induces G surface expression in Ewing sarcoma cells. EZH2 in Ewing sarcoma cells directly binds to the promoter regions of genes encoding for two key enzymes of G biosynthesis, and EZH2 inhibition enhances expression of these genes. G surface expression in Ewing sarcoma cells is not associated with distinct proliferation, colony formation, chemosensitivity, or tumorigenicity. Moreover, disruption of G synthesis by gene editing does not affect its behavior. EZH2 inhibitor treatment sensitizes Ewing sarcoma cells to effective cytolysis by G -specific CAR gene-modified T cells. In conclusion, we report a clinically applicable pharmacological approach for enhancing efficacy of adoptively transferred G -redirected T cells against Ewing sarcoma, by enabling recognition of tumor cells with low or negative target expression.