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Here we describe an innovative approach where a stable tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) chimera, referred to as membrane-bound (MB)-TRAIL, is expressed on the cell surface of MSLN-specific (M28z) CAR-T cells. This potentiates their overall anticancer activity by overcoming heterogeneous antigen expression in tumor lesions.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We engineered MB-TRAIL-armored MSLN-directed M28z CAR-T cells and assessed their in vitro toxicity using flow cytometry against tumor target cells with heterogeneous MSLN expression. Triple-multiplex bioluminescence imaging using three different reporter genes enabled simultaneous in vivo non-invasive and longitudinal assessment of CAR-T cell dynamics, as well as MSLN+ and MSLN\u2212 tumor responses in xenograft models of MSTO-211H pleural mesothelioma (PM) and A549 non-small cell-lung cancer (NSCLC) with heterogeneous antigen expression. We further tested the therapeutic benefit of combining MB-TRAIL-armored CAR-T cells with the clinically used TRAIL sensitizer, doxorubicin, in an A549 xenograft model.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>MB-TRAIL M28z CAR-T cell therapy provides pro-apoptotic stimuli to both MSLN+ and MSLN\u2212 tumors, significantly improving survival in models of MSTO-211H PM and A549 NSCLC with heterogeneous antigen expression. We also demonstrated that our treatment confers sustained protection against metastatic spread in the MSTO-211H model, whereas doxorubicin overcomes TRAIL resistance rendering the A549 model more susceptible to MB-TRAIL M28z CAR-T cell therapy and reducing metastatic growth.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>Our strategy confers superior antitumor efficacy relative to clinically trialed M28z CAR-T cells by directly targeting heterogeneous tumor antigen expression. These results constitute the foundation for developing a new class of clinically translatable TRAIL-armored CAR-T cell immunotherapies for the treatment of a wider range of malignancies.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1136\/jitc-2025-014330","type":"journal-article","created":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T15:40:37Z","timestamp":1779378037000},"page":"e014330","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":0,"title":["Membrane-bound TRAIL-armoring augments CAR-T cells in mesothelin-positive solid malignancies"],"prefix":"10.1136","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3087-9178","authenticated-orcid":false,"given":"Alessia","family":"Volpe","sequence":"first","affiliation":[{"name":"Department of Radiology, Memorial Sloan Kettering Cancer Center, New York, New York, 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