{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T15:20:56Z","timestamp":1780500056267,"version":"3.54.1"},"posted":{"date-parts":[[2023,3,10]]},"group-title":"Immunology","reference-count":50,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,3,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Generating stem memory T cells (T\n                  <jats:sub>SCM<\/jats:sub>\n                  ) is a key goal for improving cancer immunotherapy. Yet, the optimal way to modulate signaling pathways that enrich T\n                  <jats:sub>SCM<\/jats:sub>\n                  properties remains elusive. Here, we discovered that the degree to which the PI3K\u03b4 pathway is blocked pharmaceutically can generate T cells with differential levels of stemness properties. This observation was based on the progressive enrichment of transcriptional factors of stemness (Tcf-1 and Lef-1). Additional investigation revealed that T cells with high stemness features had enhanced metabolic plasticity, marked by heightened mitochondrial function and glucose uptake. Conversely, T cells with low or medium features of stemness expressed more inhibitory checkpoint receptors (Tim-3, CD39) and were vulnerable to antigen-induced cell death. Only TCR-antigen specific T cells with high stemness persisted following adoptive transfer\n                  <jats:italic>in vivo<\/jats:italic>\n                  and mounted protective immunity to melanoma tumors. Likewise, the strongest level of PI3K\u03b4 blockade\n                  <jats:italic>in vitro<\/jats:italic>\n                  generated human tumor infiltrating lymphocytes (TILs) and CAR T cells with heightened stemness properties, in turn bolstering their capacity to regress human mesothelioma tumors. We find that the level of stemness T cells possess\n                  <jats:italic>in vitro<\/jats:italic>\n                  differentially impacts their potency upon transfer in three tumor models. Mechanistically, both Lef-1 and Tcf-1 sustain anti-tumor protection by high T\n                  <jats:sub>SCM<\/jats:sub>\n                  , as deletion of either one compromised cellular therapy. Collectively, these findings highlight the therapeutic potential of carefully modulating PI3K\u03b4 signaling in T cells to confer high stemness and mediate protective responses to solid tumors.\n                <\/jats:p>","DOI":"10.1101\/2023.03.08.531589","type":"posted-content","created":{"date-parts":[[2023,3,10]],"date-time":"2023-03-10T20:10:43Z","timestamp":1678479043000},"source":"Crossref","is-referenced-by-count":3,"title":["The degree of T cell stemness differentially impacts the potency of adoptive cancer immunotherapy in a Lef-1 and Tcf-1 dependent manner"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9364-6385","authenticated-orcid":false,"given":"Guillermo O.","family":"Rangel Rivera","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2306-0570","authenticated-orcid":false,"given":"Connor J.","family":"Dwyer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0644-123X","authenticated-orcid":false,"given":"Hannah M.","family":"Knochelmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3693-4262","authenticated-orcid":false,"given":"Aubrey S.","family":"Smith","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arman","family":"Aksoy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna C.","family":"Cole","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1316-260X","authenticated-orcid":false,"given":"Megan 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