Fig. 1: G9a/GLP inhibitors improve the cytotoxicity of TCR-engineered T cells.

A Workflow for enhancing engineered TCR-T cell therapy formation using epigenetic drugs and cytotoxicity validation in 2D and 3D. Created with BioRender.com. B Cytotoxicity of T cells towards HepG2-preS1 target cells, after 5 days of treatment with a drug panel from the Structural Genomics Consortium (SGC) Open Chemistry Networks platform (Open Chem Networks) targeting epigenetic factors. Data at 48 h after co-culture of T cells and target cells are shown. Data are shown as arbitrary units of fluorescence, corresponding to cytotoxicity, following manufacturer protocol for the CellTox assay. N = 3 biologically independent donors. C Engineered TCR-T cell cytotoxicity towards the target cell line after drug treatment in a 3D cytotoxicity assay. UNC0642-treated T cells resulted in a significant increase in target cell death compared to untreated T cells (p = 0.0342). N = 3 biologically independent donors. D Representative image of 2D cell cytotoxicity assay, tracked using xCelligence. Cell index was normalized to time when T cells were added, and T cells were added at a 1:1 effector: target ratio. Cell index at E 3.5-h, F 10-h, and G 17-h after T cell addition. N = 3 biologically independent donors. H Representative images showing live (green) and dead (red) cells in 3D cytotoxicity assay, with quantification of I normalized live target cell count after addition of untreated or UNC0642-treated TCR⁺ T cells, TCR^- mock-electroporated T cells (TCR^-) and naïve T cells (no EP [electroporation]), normalized to no T cell controls and J number of invading T cells within the 3D matrix, normalized to no EP controls. TCR^- and no EP: N = 3 biologically independent donors; TCR⁺: N = 6 biologically independent donors. Data are analyzed with two-way ANOVA with matching within donors. All data are shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns not significant. Source data are provided as a Source Data file.