title: Highly compacted biodegradable DNA nanoparticles capable of overcoming the mucus barrier for inhaled lung gene therapy.
abstract: Gene therapy has emerged as an alternative for the treatment of diseases refractory to conventional therapeutics. Synthetic nanoparticle-based gene delivery systems offer highly tunable platforms for the delivery of therapeutic genes. However, the inability to achieve sustained, high-level transgene expression in vivo presents a significant hurdle. The respiratory system, although readily accessible, remains a challenging target, as effective gene therapy mandates colloidal stability in physiological fluids and the ability to overcome biological barriers found in the lung. We formulated highly stable DNA nanoparticles based on state-of-the-art biodegradable polymers, poly(β-amino esters) (PBAEs), possessing a dense corona of polyethylene glycol. We found that these nanoparticles efficiently penetrated the nanoporous and highly adhesive human mucus gel layer that constitutes a primary barrier to reaching the underlying epithelium. We also discovered that these PBAE-based mucus-penetrating DNA nanoparticles (PBAE-MPPs) provided uniform and high-level transgene expression throughout the mouse lungs, superior to several gold standard gene delivery systems. PBAE-MPPs achieved robust transgene expression over at least 4 mo following a single administration, and their transfection efficiency was not attenuated by repeated administrations, underscoring their clinical relevance. Importantly, PBAE-MPPs demonstrated a favorable safety profile with no signs of toxicity following intratracheal administration.
authors.affiliation: Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21231; Department of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21297;
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Mastorakos
authors.affiliation: Laboratory of Cellular and Molecular Physiology, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941902, Brazil;
authors.forename: Adriana L
authors.initials: AL
authors.lastname: da Silva
authors.affiliation: Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21231; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218;
authors.forename: Jane
authors.initials: J
authors.lastname: Chisholm
authors.affiliation: Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21231; Center for Biotechnology Education, Krieger School of Arts and Sciences, Johns Hopkins University, Baltimore, MD 21218;
authors.forename: Eric
authors.initials: E
authors.lastname: Song
authors.affiliation: Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21231; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218;
authors.forename: Won Kyu
authors.initials: WK
authors.lastname: Choi
authors.affiliation: Adult Cystic Fibrosis Program, Division of Pulmonary and Critical Care Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Boyle
authors.affiliation: Laboratory of Cellular and Molecular Physiology, Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, RJ 21941902, Brazil;
authors.forename: Marcelo M
authors.initials: MM
authors.lastname: Morales
authors.affiliation: Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21231; Department of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21297; Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD 21218; hanes@jhmi.edu jsuk@jhmi.edu.
authors.forename: Justin
authors.initials: J
authors.lastname: Hanes
authors.affiliation: Center for Nanomedicine, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21231; Department of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21297; hanes@jhmi.edu jsuk@jhmi.edu.
authors.forename: Jung Soo
authors.initials: JS
authors.lastname: Suk
mesh_terms: D000280:Administration, Inhalation; D000818:Animals; D003550:Cystic Fibrosis / Q000628:therapy*; D004247:DNA / Q000627:therapeutic use*; D015316:Genetic Therapy; D051379:Mice; D009093:Mucus; D053758:Nanoparticles / Q000627:therapeutic use*
chemical_list: D004247:DNA
keywords: biodegradable polymer; lung gene therapy; mucus-penetrating particles; nanotechnology; nonviral gene delivery