Abstract
Despite striking efficacy against hematologic malignancies, the cost and complexity of CAR T manufacturing present significant barriers to broader patient access. Beyond manufacturing challenges, ex vivo expansion of T cells may be detrimental to their function and persistence. Thus, delivery of CARs to reprogram host cells in vivo would represent a significant advance towards a readily available therapy, but has been limited by low efficiency, low specificity, and immunogenicity of viral vectors. Here, we describe the design of pseudotyped lentiviral vectors (LV) with superior functionality and high target specificity. We show that LV pseudotyped with chimeric envelope glycoproteins from dolphin morbillivirus (DMV) can be engineered to selectively infect human T cells and evade neutralizing antibody responses in measles-vaccinated human serum. We further demonstrate that camelid-derived nanobodies are a superior retargeting domain, overcoming limitations inherent to the use of single-chain variable fragment antibodies. Using a chimeric DMV-pseudotyped virus targeting the CD7 receptor, we demonstrate efficient and highly specific infection of T cells both in vitro and in vivo, generating functional CAR T cells and inducing therapeutic efficacy in a preclinical B cell lymphoma model.
Data availability
Data supporting the findings of this work are available within the paper and the Source Data file. The immunofluorescence data generated in this study have been deposited in the BioImage Archive under accession code S-BIAD2901. Source data are provided with this paper.
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Acknowledgements
This work was supported by the Merkin Institute for Transformative Technologies in Healthcare and Calico Life Sciences. This work was supported in part by funding from Calico Life Sciences, LLC. We thank Branka Horvat and the Measles Virus Biobank for the generous gift of the CL55 and Y503 antibodies, INSERM, France, Measles Virus Biobank https://ciri.ens-lyon.fr/teams/IbIV/measles-biobank. We thank Natan Pirete and the Broad Institute Flow Cytometry Core for their help, training, and assistance in our flow cytometry experiments, and the Broad Institute mouse facilities for providing training and assistance with in vivo experiments. We would like to thank Professor Fritz Melcher from the German Rheumatology Research Center Leibniz-Institute for providing the FGK4.5 hybridoma, Dr. Hasaya Akiba for providing the RMT3-23 hybridoma, and Dr. Yoji Murata for providing the MY-1 and Miap301 hybridomas.
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K.S.M., K.B.Y., and R.T.M. conceived the project. K.E.I., K.S.M., and P.M.A. designed and conducted the majority of experiments, analyzed experimental data, and wrote the manuscript. O.I.A., A.J.M., S.K.L.R., A.R., and N.H.K. contributed to mouse and viral experiments. C.C. conducted preliminary experiments. J.M.C. conducted liver histology and analysis and contributed to manuscript preparation. K.B.Y. and R.T.M. supervised the work and wrote the manuscript.
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R.T.M., K.B.Y., K.S.M., K.E.I., and P.M.A. are inventors on patent applications filed by the Broad Institute relating to retargeted retroviral vector technologies and the use of morbillivirus-derived and VHH-targeted vectors described in this work (U.S. Patent Application Publications US 2024/0294942 A1 and US 2024/0307555 A1). R.T.M. has received speaking or consulting fees from Bristol Myers Squibb, Gilead Sciences, Kumquat Biosciences, Immunai Therapeutics, and BioNTech and has equity ownership in OncoRev, LLC, and Jumble Therapeutics. The remaining authors declare no competing interests.
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Ibrahim, K.E., Mears, K.S., Allen, P.M. et al. In vivo CAR T cell generation using retargeted and functionalized lentiviral vectors with reduced immunogenicity. Nat Commun (2026). https://doi.org/10.1038/s41467-026-71395-y
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DOI: https://doi.org/10.1038/s41467-026-71395-y
