Research Article
Polyethyleneimine and DNA nanoparticles-based gene therapy for acute lung injury

https://doi.org/10.1016/j.nano.2013.05.004Get rights and content

Abstract

Acute lung injury (ALI) is a devastating clinical syndrome causing a substantial mortality, but to date without any effective pharmacological management in clinic. Here, we tested whether nanoparticles based on polyethylenimine (PEI) and DNA could be a potential treatment. In mouse model of ALI induced by lipopolysaccharide (LPS) (10 mg/kg), intravenous injection of PEI/DNA mediated a rapid (in 6 h) and short-lived transgene expression in lung, with alveolar epithelial cells as major targets. When β2-Adrenergic Receptor (β2AR) was applied as therapeutic gene, PEI/β2AR treatment significantly attenuated the severity of ALI, including alveolar fluid clearance, lung water content, histopathology, bronchioalveolar lavage cellularity, protein concentration, and inflammatory cytokines in mice with pre-existing ALI. In high-dose LPS (40 mg/kg)-induced ALI, post-injury treatment of PEI/β2AR significantly improved the 5-day survival of mice from 28% to 64%. These data suggest that PEI/DNA nanoparticles could be an effective agent in future clinical application for ALI treatment.

From the Clinical Editor

In this novel study, PEI/DNA nanoparticles are presented as an effective agent for the treatment of the devastating and currently untreatable syndrome of acute lung injury, using a rodent model system.

Graphical abstract

Gene therapy could be a promising approach for treatments of a variety of chronic and acute diseases. However, it has not been sufficiently applied in ALI. The vector can be a major obstacle because both viral and non-viral vectors have serious drawbacks that limit their efficacies. In this study, we showed that nanoparticles formed by PEI/DNA can deliver genes in mouse lung even in the presence of pre-existing ALI. The gene delivery is rapid, efficient, and short-lived, with alveolar epithelial cells as major targets. These properties potentiate PEI/DNA nanoparticle to be an effective therapeutic agent in the treatment of ALI.

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Section snippets

Plasmids

Luciferase and lacZ expression vectors pT3-luc and pT3-lacZ were kindly provided by Dr. Coll (INSERM-UJF U823, France).19 pcDNA3-flag-β2AR is purchased from Addgene (#14697). Plasmids were purified using Mega-prep endotoxin-free kit (Qiagen, Hilden, Germany).

Mouse Model of ALI

Five-week-old Bltw:CD1(ICR) mice were purchased from BioLasco Taiwan Co., Ltd. and maintained in Taiwan Mouse Clinic in Institute of Biomedical Sciences, Academia Sinica. All animal experiment protocols are approved by Academia Sinica

PEI/DNA-mediated gene delivery in mouse lung under healthy condition or ALI

The particles complexed by PEI and DNA were analyzed, which showed average sizes around 60 nm and zeta potentials around 30 mV (Figure S1). After intravenous injection of PEI/DNA in mice, the kinetics and target tissues of PEI/DNA-mediated gene delivery were analyzed. The reporter gene expression was found mainly in lung, where it was approximately 100-fold higher than in other organs (Figure S2). The reporter gene signal in lung was also followed and quantified using non-invasive bioluminescent

Discussion

This is the first study testing the use of PEI/DNA nanoparticles for the delivery of a therapeutic gene in the treatment of LPS-induced ALI in an animal model. Our findings suggest that the treatment with PEI/β2AR is safe in the context of LPS-induced ALI, and provides rapid and timely benefits in mice. Like previous studies,7, 8, 9, 10 our reporter gene assay showed that systemic administration of PEI/DNA nanoparticles in vivo mediated an efficient gene delivery mainly in alveolar epithelial

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    We are grateful to the animal housing and technical assistance of Taiwan Mouse Clinic (funded by the National Research Program for Biopharmaceuticals (NRPB) at NSC) and Pathology Core Lab (Institute of Biomedical Sciences, Academia Sinica). We appreciate the technical support and fruitful discussions coming from Dr. Patrick Erbacher and Dr. Jean-Luc Coll.

    We clarify that there is no conflict of interest with any financial organization regarding the material and method used and discussed in the manuscript.

    This work is supported by National Science Council (NSC) grants 100-2325-B-010-011 and 100-2321-B-010-021, and Aim for the Top University Plan of National Yang-Ming University (101ADP902).

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