Abstract
Most patients treated with US Food and Drug Administration (FDA)–approved chimeric antigen receptor (CAR) T cells eventually experience disease progression. Furthermore, CAR T cells have not been curative against solid cancers and several hematological malignancies such as T cell lymphomas, which have very poor prognoses. One of the main barriers to the clinical success of adoptive T cell immunotherapies is CAR T cell dysfunction and lack of expansion and/or persistence after infusion. In this study, we found that CD5 inhibits CAR T cell activation and that knockout (KO) of CD5 using CRISPR-Cas9 enhances the antitumor effect of CAR T cells in multiple hematological and solid cancer models. Mechanistically, CD5 KO drives increased T cell effector function with enhanced cytotoxicity, in vivo expansion, and persistence, without apparent toxicity in preclinical models. These findings indicate that CD5 is a critical inhibitor of T cell function and a potential clinical target for enhancing T cell therapies.
| Original language | English (US) |
|---|---|
| Article number | eadn6509 |
| Journal | Science Immunology |
| Volume | 9 |
| Issue number | 97 |
| DOIs | |
| State | Published - 2024 |
Funding
This work was funded through the NIH /National cancer Institute (R37-cA-262362-02), the Leukemia and Lymphoma Society, the Gilead Research Scholar Award in Hematology, the Emerson collective, the Laffey-McHugh Foundation, the Parker Institute for cancer Immunotherapy, the Berman and Maguire Funds for Lymphoma Research at Penn, and viTToria Biotherapeutics (M.R.). This work was also funded through the T32 Hematology Research Training Program and the American Society of Hematology Graduate Award (R.P.P.). Acknowledgments: We thank l. chen, m. eldabbas, and e. maddox of the human immunology core at the Perelman School of medicine at the university of Pennsylvania (nih P30 ai045008 and P30 ca016520; hic RRiD: ScR_022380). We would also like to acknowledge the university laboratory animal Resources, the cell and animal Radiation core (RRiD: ScR_022377), the Stem cell Xenograft core (RRiD: ScR_010035), the Flow cytometry core, the next-generation Sequencing core, the comparative Pathology core, and the Viral/molecular high Density Sequencing core at the university of Pennsylvania for services. Portions of Figs. 1, 4, and 7 and figs. S2 and S5 were created with BioRender.com (agreement number: Xu26WlFKBh). Funding: this work was funded through the nih /national cancer institute (R37-ca-262362-02), the leukemia and lymphoma Society, the gilead Research Scholar award in hematology, the emerson collective, the laffey-mchugh Foundation, the Parker institute for cancer immunotherapy, the Berman and maguire Funds for lymphoma Research at Penn, and vittoria Biotherapeutics (m.R.). this work was also funded through the t32 hematology Research training Program and the american Society of hematology graduate award (R.P.P.). Author contributions: R.P.P. planned and executed the experiments, analyzed the data, prepared the figures, conceptualized the study, and wrote the manuscript. g.g., y.Z., y.-h.c., W.X., P.g., K.h.K., i. chun, m.g.a., R.P., S.J.h., y.g.l., o.S., c.S., i. cohen, a.g., t.V., D.Q., S.y., a.n., a.R., o.u., m. Wang, m. Wysocka, a.c., t.B., c.X., K.g., and P.P. contributed to executing or analyzing both in vitro and in vivo experiments. their contributions spanned Dna preparation, lentiviral production, caR t cell expansion, conducting killing assays, and monitoring animals for tumor burden and blood processing and more. a.i. and J.h. contributed to the analysis and interpretation of tissue toxicity data. l.c. contributed by processing large single-cell sequencing datasets. g.g. and P.g. further contributed to preparing figures for publication. a.e.S., n.S., l.P., h.J.B., m.a.m., m. Wang, m. Wysocka, g.P.l., B.c., D.m.B., D.t.t., a.D.P., D.J.P.J., c.t.S., S.P., V.P., J.S., a.h.R., S.J.S., and S.K.B. provided essential resources, including materials and experimental/analytical support throughout the project. R.P.P. and m.R. acquired funding for this research. m.R. contributed in conceptualizing the study, contributing to the manuscript\u2019s writing, and supervising the study. Competing interests: the patent, titled \u201Cuse of cd2/5/7 knock-out anti-cd2/5/7 chimeric antigen receptor t cells against t cell lymphomas and leukemias,\u201Dhas been filed with m.R., a.D.P., and D.J.P.J. listed as coinventors, with the filing number Wo2020132327a1. the patent, titled \u201Cmethods of manufacturing t cells,\u201Dhas been filed with m.R. and n.S. listed as coinventors, with the filing number Pct/uS2024/031950. the patent, titled \u201CcD5 modified cells comprising chimeric antigen receptors (caRs) for treatment of solid tumors,\u201Dhas been filed with m.R. listed as the inventor, with the filing number Pct/ uS2022/081463. m.R. holds multiple patents related to caR t immunotherapy that are managed by the university of Pennsylvania. m.R. has served as a consultant for nanoString, BmS, gSK, Bayer, and abclon. m.R. receives research funding from abclon, Beckman coulter, and oxford nanoimaging. m.R. is the scientific founder of vittoria Biotherapeutics. R.P.P. and g.g. receive honoraria for consulting at vittoria Biotherapeutics. t.V., D.Q., S.y., a.n., a.e.S., and n.S. are employees of vittoria Biotherapeutics. n.S. is a cofounder and a member of the board of directors of vittoria Biotherapeutics. D.t.t. has patents and patents pending related to caR t. D.t.t. serves on advisory boards for Beam therapeutics, Sobi, Jazz, and Servier. D.t.t. receives research funding from Beam therapeutics and neoimmunetech. S.P. has served as Diatech Pharmacogenetics scientific board president and has served as bureau speaker for lilly, Beigene, takeda, Stemline, Kyowa Kirin, Roche, and nanoString. S.K.B. has served as a consultant to acrotech, affimed, Daiichi Sankyo, Kyowa Kirin, and Daiichi Sankyo and receives honoraria for participation in an independent data monitoring committee by J&J. S.J.S. serves on the scientific advisory boards for vittoria Biotherapeutics and caribou Biosciences. S.J.S. has received honoraria for consulting for Kite therapeutics, legend Biotech, mustang Biotech, Janssen, and novartis. S.J.S. holds multiple patents related to caR t immunotherapy that are managed by the university of Pennsylvania. all other authors declare no competing interests. Data and materials availability: all requests for raw and analyzed preclinical data and materials are promptly reviewed by the university of Pennsylvania to determine whether they are subject to intellectual property or confidentiality obligations. any data and materials that can be shared will be released via a material transfer agreement. Bulk Rna sequencing data are available from the gene expression omnibus using the accession number gSe237805. Single-cell Rna sequencing data are available from the gene expression omnibus using the accession number gSe248590. Raw data and full protein immunoblots are included in data files S1 and S2, respectively.
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology
