In vivo reprogramming of cytotoxic effector CD8+ T cells via fractalkine-conjugated mRNA-LNP

Corrigan, A. R.; Ngiow, S. F.; Statzu, M.; Pampena, M. B.; Nordin, J. M. L.; Albertus, A.; Carro, S. D.; Harper, J.; Stammen, R. L.; Wood, J. S. B.; Hamilton, J. T.; Ni, H.; Su, J.; Ramalingam, R.; Wu, V.; Paiardini, M.; Weissman, D.; Wherry, E. J.; Kreider, E.; Betts, M. R.

doi:10.1101/2025.10.29.685358

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bioRxiv · 10.1101/2025.10.29.685358

In vivo reprogramming of cytotoxic effector CD8+ T cells via fractalkine-conjugated mRNA-LNP

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Abstract

Selective in vivo reprogramming of cytotoxic effector CD8+ T (Teff) cells holds tremendous promise as a therapeutic tool but has not yet been accomplished. Here, we demonstrate that fractalkine-conjugated mRNA lipid nanoparticles (mRNA-LNP) can specifically target and deliver mRNA to CX3CR1+ Teff cells in vitro and in vivo. In mice, fractalkine-conjugated LNP target up to 90% of blood and splenic Teff cells, and delivery of IL-2-encoding mRNA to Teff cells enables robust exogenous IL-2 secretion. In rhesus macaques, fractalkine-conjugated mRNA-LNP target up to [~]100% of peripheral blood Teff cells and delivery of CD62L-mRNA enables transient CD62L expression. Collectively, these data demonstrate the potential of natural receptor ligand-based targeting of mRNA-LNP for effective and efficient transient in vivo modification of Teff cells.

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Corrigan, A. R., Ngiow, S. F., Statzu, M., Pampena, M. B., Nordin, J. M. L., Albertus, A., Carro, S. D., Harper, J., Stammen, R. L., Wood, J. S. B., Hamilton, J. T., Ni, H., Su, J., Ramalingam, R., Wu, V., Paiardini, M., Weissman, D., Wherry, E. J., Kreider, E., Betts, M. R.. 2025-10-30. In vivo reprogramming of cytotoxic effector CD8+ T cells via fractalkine-conjugated mRNA-LNP. https://doi.org/10.1101/2025.10.29.685358

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