Inhaled Nanoformulated mRNA Polyplexes for Protein Production in Lung Epithelium

Adv Mater. 2019 Feb;31(8):e1805116. doi: 10.1002/adma.201805116. Epub 2019 Jan 4.

Abstract

Noninvasive aerosol inhalation is an established method of drug delivery to the lung, and remains a desirable route for nucleic-acid-based therapeutics. In vitro transcribed (IVT) mRNA has broad therapeutic applicability as it permits temporal and dose-dependent control of encoded protein expression. Inhaled delivery of IVT-mRNA has not yet been demonstrated and requires development of safe and effective materials. To meet this need, hyperbranched poly(beta amino esters) (hPBAEs) are synthesized to enable nanoformulation of stable and concentrated polyplexes suitable for inhalation. This strategy achieves uniform distribution of luciferase mRNA throughout all five lobes of the lung and produces 101.2 ng g-1 of luciferase protein 24 h after inhalation of hPBAE polyplexes. Importantly, delivery is localized to the lung, and no luminescence is observed in other tissues. Furthermore, using an Ai14 reporter mouse model it is identified that 24.6% of the total lung epithelial cell population is transfected after a single dose. Repeat dosing of inhaled hPBAE-mRNA generates consistent protein production in the lung, without local or systemic toxicity. The results indicate that nebulized delivery of IVT-mRNA facilitated by hPBAE vectors may provide a clinically relevant delivery system to lung epithelium.

Keywords: biomaterials; gene delivery; inhalation; messenger RNA; topology.

MeSH terms

  • Administration, Inhalation
  • Animals
  • Drug Compounding / methods
  • Drug Liberation
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism*
  • Female
  • Gene Transfer Techniques
  • Genetic Therapy / methods
  • Hydrogen-Ion Concentration
  • Luciferases / genetics*
  • Lung / drug effects
  • Mice
  • Mice, Inbred C57BL
  • Models, Animal
  • Nanoparticles / chemistry*
  • Polymers / chemistry*
  • RNA, Messenger / administration & dosage
  • RNA, Messenger / adverse effects
  • RNA, Messenger / chemistry*
  • RNA, Messenger / metabolism
  • Tissue Distribution
  • Transfection / methods

Substances

  • Polymers
  • RNA, Messenger
  • poly(beta-amino ester)
  • Luciferases