期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 115, 期 26, 页码 E5859-E5866出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1805358115
关键词
gene therapy; nanoparticle; polyplex; combinatorial; immunotherapy
资金
- Department of Energy [DE-SC0018168]
- NSF [CHE-1607092]
- NIH [NIH-CA031841, NIH-CA031845]
- Stanford Center for Molecular Analysis and Design
- Child Health Research Institute
- SPARK program at Stanford University
- Norwegian Cancer Society
- NIH S10 Shared Instrument Grant [S10RR027431-01]
- U.S. Department of Energy (DOE) [DE-SC0018168] Funding Source: U.S. Department of Energy (DOE)
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 a-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.
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