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Ionizable Lipid Nanoparticle-Mediated mRNA Delivery for Human CAR T Cell Engineering
journal contribution
posted on 2020-02-05, 18:35 authored by Margaret
M. Billingsley, Nathan Singh, Pranali Ravikumar, Rui Zhang, Carl H. June, Michael J. MitchellChimeric antigen
receptor (CAR) T cell therapy relies on the ex vivo manipulation of patient T cells to create potent,
cancer-targeting therapies, shown to be capable of inducing remission
in patients with acute lymphoblastic leukemia and large B cell lymphoma.
However, current CAR T cell engineering methods use viral delivery
vectors, which induce permanent CAR expression and could lead to severe
adverse effects. Messenger RNA (mRNA) has been explored as a promising
strategy for inducing transient CAR expression in T cells to mitigate
the adverse effects associated with viral vectors, but it most commonly
requires electroporation for T cell mRNA delivery, which can be cytotoxic.
Here, ionizable lipid nanoparticles (LNPs) were designed for ex vivo mRNA delivery to human T cells. A library of 24
ionizable lipids was synthesized, formulated into LNPs, and screened
for luciferase mRNA delivery to Jurkat cells, revealing seven formulations
capable of enhanced mRNA delivery over lipofectamine. The top-performing
LNP formulation, C14–4, was selected for CAR mRNA delivery
to primary human T cells. This platform induced CAR expression at
levels equivalent to electroporation, with substantially reduced cytotoxicity.
CAR T cells engineered via C14–4 LNP treatment were then compared
to electroporated CAR T cells in a coculture assay with Nalm-6 acute
lymphoblastic leukemia cells, and both CAR T cell engineering methods
elicited potent cancer-killing activity. These results demonstrate
the ability of LNPs to deliver mRNA to primary human T cells to induce
functional protein expression, and indicate the potential of LNPs
to enhance mRNA-based CAR T cell engineering methods.
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24 ionizable lipidsmRNA-based CAR T cell engineering methodsCAR T cell engineering methods useT cell mRNA deliveryCAR T cellselectroporated CAR T cellsT cell therapylymphoblastic leukemia cellsionizable lipid nanoparticlesluciferase mRNA deliverypatient T cellsLNPB cell lymphomaCAR mRNA deliveryvivo mRNA deliveryCAR expressionIonizable Lipid Nanoparticle-Mediated mRNA DeliveryHuman CAR T Cell Engineering Chimeric antigen receptorT cellsCAR T cell engineering methods
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