Encapsulating Polyethyleneimine-DNA Nanoplexes into PEGylated Biodegradable Microparticles Increases Transgene Expression In Vitro and Reduces Inflammatory Responses In Vivo

Treniece L. Terry, Brittany E. Givens, Andrea Adamcakova-Dodd, Peter S. Thorne, Victor G.J. Rodgers, Aliasger K. Salem

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Encapsulating genetic material into biocompatible polymeric microparticles is a means to improving gene transfection while simultaneously decreasing the tendency for inflammatory responses; and can be advantageous in terms of delivering material directly to the lungs via aerosolization for applications such as vaccinations. In this study, we investigated the advantages of using polymeric microparticles carrying the luciferase reporter gene in increasing transfection efficiency in the readily transfectable HEK293 cell line and the difficult to transfect RAW264.7 cell line. The results indicated that there was a limit to the ratio of nitrogen in polyethylenimine (PEI) to phosphate in DNA (N/P ratio) beyond which further increases in transgene expression no longer, or only marginally, occurred. Microparticles encapsulating PEI:DNA nanoplexes induced cellular toxicity in a dose-dependent manner. PEGylation increased transgene expression, likely related to enhanced degradation of particles. Furthermore, intra-tracheal instillation in rats allowed us to investigate the inflammatory response in the lung as a function of PEGylation, porosity, and size. Porosity did not influence cell counts in bronchoalveolar lavage fluid in the absence of PEG, but in particles containing PEG, non-porous particles recruited fewer inflammatory cells than their porous counterparts. Finally, both 1 μm and 10 μm porous PLA-PEG particles recruited more neutrophils than 4 μm particles. Thus, we have shown that PEGylation and lack of porosity are advantageous for faster release of genetic cargo from microparticles and a reduced inflammatory response, respectively.

Original languageEnglish
Article number69
JournalAAPS PharmSciTech
Volume22
Issue number2
DOIs
StatePublished - Feb 2021

Bibliographical note

Funding Information:
Histology was performed in the Central Microscopy Research Facility (CMRF) at the University of Iowa. In vivo studies were conducted in laboratory facilities supported by the Environmental Health Sciences Research Center funded by NIH P30 ES005605.

Funding Information:
Histology was performed in the Central Microscopy Research Facility (CMRF) at the University of Iowa. In vivo studies were conducted in laboratory facilities supported by the Environmental Health Sciences Research Center funded by NIH P30 ES005605.

Funding Information:
A.K.S acknowledges support from the NIH P30 CA086862 grant and the Lyle and Sharon Bighley Chair of Pharmaceutical Sciences. T.L.T. acknowledges support from the Department of Education GAANN Fellowship program. B.E.G. acknowledges funding support from the Alfred P. Sloan Foundation, the University of Iowa Graduate College, and the National GEM Consortium. V. G. J. R. acknowledges support from the Jacques S. Yeager, Sr. endowment.

Publisher Copyright:
© 2021, American Association of Pharmaceutical Scientists.

Keywords

  • Gene transfection
  • PEGylation
  • PLA-PEG
  • PLGA
  • Porosity

ASJC Scopus subject areas

  • Pharmaceutical Science

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