Directed evolution-based discovery of ligands for in vivo restimulation of chimeric antigen receptor T cells
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s41551-025-01470-0.pdf
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Published version
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15.72 MB
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Checksum (MD5)
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Author(s)
Grzywa, Tomasz M
Neeser, Alexandra
Ramasubramanian, Ranjani
Romanov, Anna
Tannir, Ryan
Mehta, Naveen K
Cossette, Benjamin
Morgan, Duncan M
Goncalves, Beatriz
Sukaj, Ina
Date Issued
August 25, 2025
Journal
Nature Biomedical Engineering
Publisher
Springer Science and Business Media LLC
Citation
Grzywa, T.M., Neeser, A., Ramasubramanian, R. et al. Directed evolution-based discovery of ligands for in vivo restimulation of chimeric antigen receptor T cells. Nat. Biomed. Eng (2025).
Version
Final published version
Abstract
Chimeric antigen receptor (CAR) T cell therapy targeting CD19 elicits remarkable clinical efficacy in B cell malignancies, but many patients relapse owing to failed expansion and/or progressive loss of CAR-T cells. We recently reported a strategy to potently restimulate CAR-T cells in vivo, enhancing their functionality by administration of a vaccine-like stimulus comprised of surrogate peptide ligands for a CAR linked to a lymph node-targeting amphiphilic PEG-lipid (amph-vax). Here we demonstrate a general strategy to discover and optimize peptide mimotopes enabling amph-vax generation for any CAR. We use yeast surface display to identify peptide binders to FMC63 (the scFv used in clinical CD19 CARs), which are then subsequently affinity matured by directed evolution. CAR-T vaccines using these optimized mimotopes triggered marked expansion and memory development of CD19 CAR-T cells in both syngeneic and humanized mouse models of B-acute lymphoblastic leukaemia/lymphoma, and enhanced control of disease progression compared with CD19 CAR-T-only-treated mice. This approach enables amph-vax boosting to be applied to any clinically relevant CAR-T cell product.
MIT Department
Koch Institute for Integrative Cancer Research at MIT
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Chemical Engineering
Ragon Institute of MGH, MIT and Harvard
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DOI of Published Version
10.1038/s41551-025-01470-0
