Research ArticlePolyethyleneimine and DNA nanoparticles-based gene therapy for acute lung injury
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.
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
References (45)
- et al.
Effects of beta2-adrenergic receptor overexpression on alveolar epithelial active transport
J Allergy Clin Immunol
(2002) - et al.
Lifelong reporter gene imaging in the lungs of mice following polyethyleneimine-mediated sleeping-beauty transposon delivery
Biomaterials
(2011) - et al.
MAD2B, a novel TCF4-binding protein, modulates TCF4-mediated epithelial–mesenchymal transdifferentiation
J Biol Chem
(2009) Beta-agonists for ARDS: the dark side of adrenergic stimulation?
Lancet
(2012)- et al.
Endocytosis of nanomedicines
J Control Release
(2010) - et al.
Biodistribution and tissue expression kinetics of plasmid DNA complexed with polyethylenimines of different molecular weight and structure
J Control Release
(2007) - et al.
Endotoxin removal from protein solutions
J Biotechnol
(2000) - et al.
Modulation of eIF5A expression using SNS01 nanoparticles inhibits NF-kappaB activity and tumor growth in murine models of multiple myeloma
Mol Ther
(2012) - et al.
The acute respiratory distress syndrome: pathogenesis and treatment
Annu Rev Pathol
(2011) - et al.
Pharmacotherapy of acute lung injury and acute respiratory distress syndrome
Curr Med Chem
(2008)
Incidence and outcomes of acute lung injury
N Engl J Med
Effect of intravenous beta-2 agonist treatment on clinical outcomes in acute respiratory distress syndrome (BALTI-2): a multicentre, randomised controlled trial
Lancet
Randomized, placebo-controlled clinical trial of an aerosolized beta-2 agonist for treatment of acute lung injury
Am J Respir Crit Care Med
Non-viral and viral vectors for gene therapy
Cell Mol Biol
Polyethylenimine-based intravenous delivery of transgenes to mouse lung
Gene Ther
Systemic linear polyethylenimine (L-PEI)-mediated gene delivery in the mouse
J Gene Med
Comparison between cationic polymers and lipids in mediating systemic gene delivery to the lungs
Gene Ther
Systemic delivery of DNA or sirna mediated by linear polyethylenimine (L-PEI) does not induce an inflammatory response
Pharm Res
Alveolar epithelial beta2-adrenergic receptors
Am J Respir Cell Mol Biol
In vitro and in vivo effects of salbutamol on neutrophil function in acute lung injury
Thorax
Antiinflammatory effects of salmeterol after inhalation of lipopolysaccharide by healthy volunteers
Am J Respir Crit Care Med
In vivo and in vitro effects of salbutamol on alveolar epithelial repair in acute lung injury
Thorax
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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).