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However, rapamycin has potential off-target and side effects that warrant the discovery of additional TOR inhibitors. TOR was initially discovered in <jats:italic>Saccharomyces cerevisiae<\/jats:italic> (yeast) which contains two TOR paralogs, <jats:italic>TOR1<\/jats:italic> and <jats:italic>TOR2<\/jats:italic>. Yeast lacking functional Tor1 are viable but are hypersensitive to growth inhibition by TORC1 inhibitors, which is a property of yeast that can be exploited to identify TOR inhibitors. Additionally, yeast lacking FK506-sensitive proline rotamase (<jats:italic>FPR1<\/jats:italic>) or containing a <jats:italic>tor1-1<\/jats:italic> allele (a mutation in the Fpr1-rapamycin binding domain of Tor1) are robustly and selectively resistant to rapamycin and analogs that allosterically inhibit TOR activity via an <jats:italic>FPR1<\/jats:italic>-dependent mechanism. To facilitate the identification of TOR inhibitors, we generated a panel of yeast strains with mutations in TOR pathway genes combined with the removal of 12 additional genes involved in drug efflux. This creates a drug-sensitive strain background that can sensitively and effectively identify TOR inhibitors. In a wild-type yeast strain background, 25\u00a0\u00b5M of Torin1 and 100\u00a0\u00b5M of GSK2126458 (omipalisib) are necessary to observe <jats:italic>TOR1<\/jats:italic>-dependent growth inhibition by these known TOR inhibitors. In contrast, 100\u00a0nM Torin1 and 500\u00a0nM GSK2126458 (omipalisib) are sufficient to identify <jats:italic>TOR1-<\/jats:italic>dependent growth inhibition in the drug-sensitized background. This represents a 200-fold and 250-fold increase in detection sensitivity for Torin1 and GSK2126458, respectively. Additionally, for the TOR inhibitor AZD8055, the drug-sensitive system resolves that the compound results in <jats:italic>TOR1-<\/jats:italic>dependent growth sensitivity at 100\u00a0\u00b5M, whereas no growth inhibition is observed in a wild-type yeast strain background. Our platform also identifies the caffeine analog aminophylline as a <jats:italic>TOR1<\/jats:italic>-dependent growth inhibitor via selective <jats:italic>tor1<\/jats:italic> growth sensitivity. We also tested nebivolol, isoliquiritigenin, canagliflozin, withaferin A, ganoderic acid A, and taurine and found no evidence for TOR inhibition using our yeast growth-based model. Our results demonstrate that this system is highly effective at identifying compounds that inhibit the TOR pathway. It offers a rapid, cost-efficient, and sensitive tool for drug discovery, with the potential to expedite the identification of new TOR inhibitors that could serve as geroprotective and\/or anti-cancer agents.<\/jats:p>","DOI":"10.1007\/s11357-025-01534-8","type":"journal-article","created":{"date-parts":[[2025,1,30]],"date-time":"2025-01-30T18:17:21Z","timestamp":1738261041000},"page":"5605-5617","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["An mTOR inhibitor discovery system using drug-sensitized yeast"],"prefix":"10.1007","volume":"47","author":[{"given":"Anna K.","family":"Breen","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sarah","family":"Thomas","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David","family":"Beckett","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew","family":"Agsalud","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Graham","family":"Gingras","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Judd","family":"Williams","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7090-086X","authenticated-orcid":false,"given":"Brian M.","family":"Wasko","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,1,30]]},"reference":[{"key":"1534_CR1","doi-asserted-by":"publisher","first-page":"2245","DOI":"10.18632\/aging.101319","volume":"9","author":"A Aliper","year":"2017","unstructured":"Aliper A, Jellen L, Cortese F, et al. 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