title: N-acetylcysteine enhances cystic fibrosis sputum penetration and airway gene transfer by highly compacted DNA nanoparticles.
abstract: For effective airway gene therapy of cystic fibrosis (CF), inhaled gene carriers must first penetrate the hyperviscoelastic sputum covering the epithelium. Whether clinically studied gene carriers can penetrate CF sputum remains unknown. Here, we measured the diffusion of a clinically tested nonviral gene carrier, composed of poly-l-lysine conjugated with a 10 kDa polyethylene glycol segment (CK(30)PEG(10k)). We found that CK(30)PEG(10k)/DNA nanoparticles were trapped in CF sputum. To improve gene carrier diffusion across sputum, we tested adjuvant regimens consisting of N-acetylcysteine (NAC), recombinant human DNase (rhDNase) or NAC together with rhDNase. While rhDNase alone did not enhance gene carrier diffusion, NAC and NAC + rhDNase increased average effective diffusivities by 6-fold and 13-fold, respectively, leading to markedly greater fractions of gene carriers that may penetrate sputum layers. We further tested the adjuvant effects of NAC in the airways of mice with Pseudomonas aeruginosa lipopolysaccharide (LPS)-induced mucus hypersecretion. Intranasal dosing of NAC prior to CK(30)PEG(10k)/DNA nanoparticles enhanced gene expression by up to ~12-fold compared to saline control, reaching levels observed in the lungs of mice without LPS challenge. Our findings suggest that a promising synthetic nanoparticle gene carrier may transfer genes substantially more effectively to lungs of CF patients if administered following adjuvant mucolytic therapy with NAC or NAC + rhDNase.
authors.affiliation: Department of Ophthalmology, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21231, USA.
authors.forename: Jung Soo
authors.initials: JS
authors.lastname: Suk
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Boylan
authors.forename: Kanika
authors.initials: K
authors.lastname: Trehan
authors.forename: Benjamin C
authors.initials: BC
authors.lastname: Tang
authors.forename: Craig S
authors.initials: CS
authors.lastname: Schneider
authors.forename: Jung-Ming G
authors.initials: JM
authors.lastname: Lin
authors.forename: Michael P
authors.initials: MP
authors.lastname: Boyle
authors.forename: Pamela L
authors.initials: PL
authors.lastname: Zeitlin
authors.forename: Samuel K
authors.initials: SK
authors.lastname: Lai
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Cooper
authors.forename: Justin
authors.initials: J
authors.lastname: Hanes
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D000328:Adult; D000818:Animals; D001704:Biopolymers / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000628:therapy; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004058:Diffusion / Q000187:drug effects; D005100:Expectorants / Q000494:pharmacology*; D005260:Female; D015316:Genetic Therapy; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009077:Mucins / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry / Q000378:metabolism; D011107:Polylysine / Q000737:chemistry / Q000378:metabolism; D012137:Respiratory System / Q000187:drug effects / Q000378:metabolism; D013183:Sputum / Q000187:drug effects*; D014161:Transduction, Genetic / Q000379:methods*; D014783:Viscosity / Q000187:drug effects; D055815:Young Adult
chemical_list: D001704:Biopolymers; D005100:Expectorants; D009077:Mucins; D011107:Polylysine; D011092:Polyethylene Glycols; D004247:DNA; D000111:Acetylcysteine