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bioRxiv · 10.64898/2026.02.18.706661

Self-amplifying RNA-based CAR T cell therapy with enhanced duration and multi-genic logic function

Gu, Y.·Choi, J.·Mutha, D.·Wu, C.·Ganem, N. J.·Grinstaff, M.·Wong, W.
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Abstract

Chimeric antigen receptor T (CAR-T) cell therapy is transforming the treatment landscape of hematological malignancies. However, manufacturing with integrating viral vectors is costly, slow, and carries risks including insertional mutagenesis, prolonged B cell aplasia, and other long-term toxicities. Expression of CAR with mRNA can reduce cost, manufacturing timelines, and improve safety. However, the short-lived expression necessitates frequent repeat dosing. Here, we describe a modified self-amplifying RNA (saRNA) platform for engineering CAR T cells with prolonged CAR expression and enhanced durability of tumor control relative to mRNA CAR T cells. In an acute lymphoblastic leukemia (ALL) xenograft model, saRNA CAR T cells achieve superior tumor suppression and prolong survival. Further, a single-strand modified saRNA supports the co-expression of multiple proteins, enabling the construction of advanced CAR systems, such as OR- and AND-gated logic CAR T cells. Together, these results highlight saRNA as a powerful and versatile platform for CAR T cell engineering with favorable safety, efficacy, and accessibility.

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Gu, Y., Choi, J., Mutha, D., Wu, C., Ganem, N. J., Grinstaff, M., Wong, W.. 2026-02-20. Self-amplifying RNA-based CAR T cell therapy with enhanced duration and multi-genic logic function. https://doi.org/10.64898/2026.02.18.706661

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