Lipophilic Polycation Vehicles Display High Plasmid DNA Delivery to Multiple Cell Types

Yaoying Wu, Adam E. Smith, Theresa M. Reineke

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

A class of cationic poly(alkylamidoamine)s (PAAAs) containing lipophilic methylene linkers were designed and examined as in vitro plasmid DNA (pDNA) delivery agents. The PAAAs were synthesized via step-growth polymerization between a diamine monomer and each of four different diacid chloride monomers with varying methylene linker lengths, including glutaryl chloride, adipoyl chloride, pimeloyl chloride, and suberoyl chloride, which served to systematically increase the lipophilicity of the polymers. The synthesized polymers successfully complexed with pDNA in reduced serum medium at N/P ratios of 5 and greater, resulting in polyplexes with hydrodynamic diameters of approximately 1 μm. These polyplexes were tested for in vitro transgene expression and cytotoxicity using HDFa (human dermal fibroblast), HeLa (human cervical carcinoma), HMEC (human mammary epithelial), and HUVEC (human umbilical vein endothelial) cells. Interestingly, select PAAA polyplex formulations were found to be more effective than Lipofectamine 2000 at promoting transgene expression (GFP) while maintaining comparable or higher cell viability. Transgene expression was highest in HeLa cells (∼90% for most formulations) and lowest in HDFa cells (up to ∼20%) as measured by GFP fluorescence. In addition, the cytotoxicity of PAAA polyplex formulations was significantly increased as the molecular weight, N/P ratio, and methylene linker length were increased. The PAAA vehicles developed herein provide a new delivery vehicle design strategy of displaying attributes of both polycations and lipids, which show promise as a tunable scaffold for refining the structure-activity-toxicity profiles for future genome editing studies.

Original languageEnglish (US)
Pages (from-to)2035-2040
Number of pages6
JournalBioconjugate Chemistry
Volume28
Issue number8
DOIs
StatePublished - Aug 16 2017

Bibliographical note

Funding Information:
We thank Young Joon Kim and Luis Ugozzoli at Bio-Rad, Inc. for their support of the flow cytometry experiments and the National Institutes of Health (NIH) Directors New Innovator Award Program (DP2OD006669-01) and Bio-Rad, Inc. for financial support of this project.

Publisher Copyright:
© 2017 American Chemical Society.

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