Enhanced mRNA delivery into lymphocytes enabled by lipid-varied libraries of charge-altering releasable transporters.

Autor:	McKinlay CJ; Department of Chemistry, Stanford University, Stanford, CA 94305., Benner NL; Department of Chemistry, Stanford University, Stanford, CA 94305., Haabeth OA; Division of Oncology, Department of Medicine, Stanford Cancer Institute, Stanford University, Stanford, CA 94305., Waymouth RM; Department of Chemistry, Stanford University, Stanford, CA 94305; waymouth@stanford.edu wenderp@stanford.edu., Wender PA; Department of Chemistry, Stanford University, Stanford, CA 94305; waymouth@stanford.edu wenderp@stanford.edu.; Department of Chemical and Systems Biology, Stanford University, Stanford, CA 94305.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2018 Jun 26; Vol. 115 (26), pp. E5859-E5866. Date of Electronic Publication: 2018 Jun 11.
DOI:	10.1073/pnas.1805358115
Abstrakt:	We report a strategy for generating a combinatorial library of oligonucleotide transporters with varied lipid domains and their use in the efficient transfection of lymphocytes with mRNA in vitro and in vivo. This library is based on amphiphilic charge-altering releasable transporters (CARTs) that contain a lipophilic block functionalized with various side-chain lipids and a polycationic α-amino ester mRNA-binding block that undergoes rearrangement to neutral small molecules, resulting in mRNA release. We show that certain binary mixtures of these lipid-varied CARTs provide up to a ninefold enhancement in mRNA translation in lymphocytes in vitro relative to either a single-lipid CART component alone or the commercial reagent Lipofectamine 2000, corresponding to a striking increase in percent transfection from 9-12% to 80%. Informed by the results with binary mixtures, we further show that CARTs consisting of optimized ratios of the two lead lipids incorporated into a single hybrid-lipid transporter molecule maintain the same delivery efficacy as the noncovalent mixture of two CARTs. The lead lipid CART mixtures and hybrid-lipid CARTs show enhanced lymphocyte transfection in primary T cells and in vivo in mice. This combinatorial approach for rapidly screening mRNA delivery vectors has provided lipid-varied CART mixtures and hybrid-lipid CARTs that exhibit significant improvement in mRNA delivery to lymphocytes, a finding of potentially broad value in research and clinical applications. Competing Interests: The authors declare no conflict of interest.
Databáze:	MEDLINE
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