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Stereoselective Degradation of Diacylglycerol Kinases Potentiate T cell Activation and Tumor Cell Cytotoxicity

Shaikh, M.; Mookherjee, S. P.; Weckerly, C.; Libby, A. H.; Xiao, A.; Zhao, Y.; Vaidya, S. D.; Kim, A.; Li, Z.; Ware, M. L.; Marants, M.; Murtagh, O.; Wolfe, W. J.; Bullock, T. N.; Purow, B. W.; Hammond, G. R.; Hsu, K.

2025-12-12 biochemistry
10.64898/2025.12.09.692983 bioRxiv
Show abstract

Stereoselective recognition is a powerful means to differentiate selective versus non-specific activity of small molecules in complex biological systems. Here, we disclose stereochemically defined, sulfonyl-triazole inhibitors of the lipid enzyme diacylglycerol kinase-alpha (DGK), a key metabolic checkpoint for T cell effector function. Acute treatment with the covalent DGK inhibitor AHL-7160 recruited endogenous DGK to the plasma membrane in a stereoselective and isozyme-specific manner. The membrane translocation activity of AHL-7160 correlated with blockade of cellular phosphatidic acid production and potentiation of primary T cell-mediated killing of a glioblastoma cell line. Quantitative chemoproteomics revealed Y669 and K411 as sites of AHL-7160 modification on endogenous DGK in cells. Extended treatments resulted in proteasome-dependent and proteome-wide selective degradation of DGK in T cells. Collectively, these findings establish covalent DGK ligands as potent molecular glues with translational potential in immunotherapy.

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