Journal article
The absence of CpG in plasmid DNA-chitosan polyplexes enhances transfection efficiencies and reduces inflammatory responses in murine lungs
Molecular pharmaceutics, Vol.11(3), pp.1022-1031
03/03/2014
DOI: 10.1021/mp400689r
PMCID: PMC3993893
PMID: 24494979
Abstract
Chitosan polyplexes containing plasmid DNA (pDNA) have significant potential for pulmonary gene delivery applications. However, prior to using chitosan/pDNA polyplexes (CSpp) in clinical applications, their potential cytotoxicity needs to be investigated. In this study, we formulated 200-400 nm CSpp with amine to phosphate (N/P) ratios that ranged from 1 to 100. We compared two types of plasmids within CSpp: pDNA that was free of CpG sequences (CpG(-)) and pDNA that contained CpG sequences (CpG(+)). Both forms of CSpp showed low cytotoxicity when cultured with A549 and HEK293 cell lines in vitro. CSpp(CpG(-)) generated higher luciferase expression both in vitro, for A549 cells, and in vivo, compared with CSpp(CpG(+)). In addition, CSpp(CpG(-)) elicited milder inflammatory responses in mice one day subsequent to nasal instillation, as determined by proinflammatory cytokine levels within the bronchoalveolar lavage fluid. Our findings suggest that to achieve optimal gene expression with minimal cytotoxicity, inflammation, and oxidative stress, the N/P ratios and CpG sequences in the pDNA of CSpp need to be considered. These findings will inform the preclinical safety assessments of CSpp in pulmonary gene delivery systems.
Details
- Title: Subtitle
- The absence of CpG in plasmid DNA-chitosan polyplexes enhances transfection efficiencies and reduces inflammatory responses in murine lungs
- Creators
- Amaraporn Wongrakpanich - Division of Pharmaceutics and Translational Therapeutics, College of Pharmacy, ‡Department of Occupational and Environmental Health, College of Public Health, and §Department of Radiation Oncology, Carver College of Medicine, University of Iowa , Iowa City, Iowa 52242, United StatesAndrea Adamcakova-DoddWei XieVijaya B JoshiKranti A MapuskarSean M GearyDouglas R SpitzPeter S ThorneAliasger K Salem
- Resource Type
- Journal article
- Publication Details
- Molecular pharmaceutics, Vol.11(3), pp.1022-1031
- DOI
- 10.1021/mp400689r
- PMID
- 24494979
- PMCID
- PMC3993893
- NLM abbreviation
- Mol Pharm
- ISSN
- 1543-8392
- eISSN
- 1543-8392
- Publisher
- United States
- Grant note
- P30 ES005605 / NIEHS NIH HHS 1R21CA13345-01 / NCI NIH HHS NIH P30ES005605 / NIEHS NIH HHS 1R21CA128414-01A2/UI / NCI NIH HHS R21 CA128414 / NCI NIH HHS P30-CA086862 / NCI NIH HHS P30 DK054759 / NIDDK NIH HHS P30 CA086862 / NCI NIH HHS
- Language
- English
- Date published
- 03/03/2014
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Civil and Environmental Engineering; Occupational and Environmental Health; Stead Family Department of Pediatrics; Pathology; Pharmaceutical Sciences and Experimental Therapeutics; Radiation Oncology; Craniofacial Anomalies Research Center; Dental Research; Chemical and Biochemical Engineering
- Record Identifier
- 9983985964602771
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