Lung gene therapy with highly compacted DNA nanoparticles that overcome the mucus barrier.
Inhaled gene carriers must penetrate the highly viscoelastic and adhesive mucus barrier in the airway in order to overcome rapid mucociliary clearance and reach the underlying epithelium; however, even the most widely used viral gene carriers are unable to efficiently do so. We developed two polymeric gene carriers that compact plasmid DNA into small and highly stable nanoparticles with dense polyethylene glycol (PEG) surface coatings. These highly compacted, densely PEG-coated DNA nanoparticles rapidly penetrate human cystic fibrosis (CF) mucus ex vivo and mouse airway mucus ex situ. Intranasal administration of the mucus penetrating DNA nanoparticles greatly enhanced particle distribution, retention and gene transfer in the mouse lung airways compared to conventional gene carriers. Successful delivery of a full-length plasmid encoding the cystic fibrosis transmembrane conductance regulator protein was achieved in the mouse lungs and airway cells, including a primary culture of mucus-covered human airway epithelium grown at air-liquid interface, without causing acute inflammation or toxicity. Highly compacted mucus penetrating DNA nanoparticles hold promise for lung gene therapy.
The Center for Nanomedicine, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA; Department of Ophthalmology, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA; Department of Biomedical Engineering, The Johns Hopkins University School of Medicine, 720 Rutland Avenue, Baltimore 21205, USA.
Jung Soo
JS
Suk
The Center for Nanomedicine, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA.
Anthony J
AJ
Kim
The Center for Nanomedicine, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA; Department of Molecular & Cellular Biology, The Johns Hopkins University, 3400 North Charles Street, Baltimore 21218, USA.
Kanika
K
Trehan
The Center for Nanomedicine, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA; Department of Chemical & Biomolecular Engineering, The Johns Hopkins University, 3400 North Charles Street, Baltimore 21218, USA.
Craig S
CS
Schneider
The Center for Nanomedicine, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA; Department of Ophthalmology, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA.
Liudmila
L
Cebotaru
Department of Physiology, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore 21205, USA.
Owen M
OM
Woodward
Department of Chemical & Biomolecular Engineering, The Johns Hopkins University, 3400 North Charles Street, Baltimore 21218, USA.
Nicholas J
NJ
Boylan
Johns Hopkins Adult Cystic Fibrosis Program, Division of Pulmonary and Critical Care Medicine, The Johns Hopkins University School of Medicine, 1830 East Monument Street, Baltimore 21205, USA.
Michael P
MP
Boyle
The Center for Nanomedicine, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA; Department of Chemical & Biomolecular Engineering, The Johns Hopkins University, 3400 North Charles Street, Baltimore 21218, USA.
Samuel K
SK
Lai
The Center for Nanomedicine, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA; Department of Physiology, The Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore 21205, USA.
William B
WB
Guggino
The Center for Nanomedicine, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA; Department of Ophthalmology, The Johns Hopkins University School of Medicine, 400 North Broadway, Baltimore 21231, USA; Department of Biomedical Engineering, The Johns Hopkins University School of Medicine, 720 Rutland Avenue, Baltimore 21205, USA; Department of Chemical & Biomolecular Engineering, The Johns Hopkins University, 3400 North Charles Street, Baltimore 21218, USA. Electronic address: hanes@jhmi.edu.
Justin
J
Hanes
D000281:Administration, Intranasal; D000328:Adult; D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D003550:Cystic Fibrosis / Q000378:metabolism / Q000628:therapy; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry; D004947:Esophagus / Q000378:metabolism; D005260:Female; D005753:Gastric Mucosa / Q000378:metabolism; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006801:Humans; D008168:Lung / Q000378:metabolism*; D008807:Mice, Inbred BALB C; D009093:Mucus / Q000378:metabolism*; D053758:Nanoparticles / Q000008:administration & dosage* / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011094:Polyethyleneimine / Q000737:chemistry; D011107:Polylysine / Q000737:chemistry; D055815:Young Adult
D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D011107:Polylysine; D011092:Polyethylene Glycols; D011094:Polyethyleneimine; D004247:DNA
Airway gene therapy; DNA nanoparticle; Mucus barrier; Multiple particle tracking