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Gene perturbation studies suggest that inhibiting TOX may have therapeutic implications for both leukemia and T cell exhaustion. However, due to its complex molecular mechanisms and intrinsically disordered structure, TOX has not been effectively targeted by small molecules to date. In this study, we used small molecule microarray (SMM) screening and biochemical assays to identify a series of TOX protein-protein interaction (PPI) inhibitors. We identified\n                  <jats:bold>KI-TOX-A3<\/jats:bold>\n                  as a TOX protein binder and potent TOX PPI inhibitor. In T-ALL,\n                  <jats:bold>KI-TOX-A3<\/jats:bold>\n                  revealed selective cytotoxicity and proteosome-dependent TOX degradation. In CD8\n                  <jats:sup>+<\/jats:sup>\n                  T cells,\n                  <jats:bold>KI-TOX-A3<\/jats:bold>\n                  potently reversed T cell exhaustion by decreasing surface inhibitory receptors, increasing expression of effector cytokines, and enhancing cancer cell killing activity. We also demonstrate the utility of\n                  <jats:bold>KI-TOX-A3<\/jats:bold>\n                  to probe potential epigenetic regulatory mechanisms of TOX via KAT7 acetylation in T cells.\n                <\/jats:p>","DOI":"10.1101\/2025.03.03.641115","type":"posted-content","created":{"date-parts":[[2025,3,10]],"date-time":"2025-03-10T13:55:19Z","timestamp":1741614919000},"source":"Crossref","is-referenced-by-count":1,"title":["Small molecule modulators of TOX protein re-invigorate T cell activity"],"prefix":"10.64898","author":[{"given":"Bocheng","family":"Wu","sequence":"first","affiliation":[{"name":"Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge MA 02139"},{"name":"Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA 02142"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heng Jui","family":"Chang","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge MA 02139"},{"name":"Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA 02142"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Prashant","family":"Singh","sequence":"additional","affiliation":[{"name":"Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA 02142"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander","family":"Hostetler","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge MA 02139"},{"name":"Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139"},{"name":"Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yichen","family":"Xiang","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge MA 02139"},{"name":"Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shenghao","family":"Guo","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge MA 02139"},{"name":"Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fiona Yihan","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia J.","family":"Zhong","sequence":"additional","affiliation":[{"name":"Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Becky S.","family":"Leifer","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge MA 02139"},{"name":"Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA 02142"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard P.","family":"Schiavoni","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nan","family":"Jiang","sequence":"additional","affiliation":[{"name":"Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge MA 02139"},{"name":"Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA 02142"},{"name":"Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amit","family":"Choudhary","sequence":"additional","affiliation":[{"name":"Broad Institute of Massachusetts Institute of Technology and Harvard University, Cambridge, MA 02142"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter M. 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