Lipid nanoparticle formulations for optimal RNA-based topical delivery to murine airways.

Autor: Tam A; NanoVation Therapeutics Inc. Vancouver, British Columbia, Canada; Michael Cuccione Childhood Cancer Research Program, BC Children's Hospital Research Institute, University of British Columbia Vancouver, British Columbia, Canada; University of British Columbia (UBC) Center for Heart Lung Innovation, St. Paul's Hospital, Vancouver, British Columbia, Canada., Kulkarni J; NanoVation Therapeutics Inc. Vancouver, British Columbia, Canada; NanoMedicines Innovation Network, Vancouver, British Columbia, Canada; University of British Columbia (UBC), Department of Biochemistry and Molecular Biology, Vancouver, British Columbia, Canada., An K; NanoVation Therapeutics Inc. Vancouver, British Columbia, Canada; University of British Columbia (UBC), Department of Biochemistry and Molecular Biology, Vancouver, British Columbia, Canada., Li L; Michael Cuccione Childhood Cancer Research Program, BC Children's Hospital Research Institute, University of British Columbia Vancouver, British Columbia, Canada., Dorscheid DR; University of British Columbia (UBC) Center for Heart Lung Innovation, St. Paul's Hospital, Vancouver, British Columbia, Canada., Singhera GK; University of British Columbia (UBC) Center for Heart Lung Innovation, St. Paul's Hospital, Vancouver, British Columbia, Canada; Department of Medicine (Division of Respirology), UBC, Vancouver, British Columbia, Canada., Bernatchez P; University of British Columbia (UBC) Center for Heart Lung Innovation, St. Paul's Hospital, Vancouver, British Columbia, Canada; Department of Medicine (Division of Respirology), UBC, Vancouver, British Columbia, Canada; Department of Anesthesiology, Pharmacology & Therapeutics, University of British Columbia, 217-2176 Health Sciences Mall, Vancouver, British Columbia V6T 1Z3, Canada., Reid G; Michael Cuccione Childhood Cancer Research Program, BC Children's Hospital Research Institute, University of British Columbia Vancouver, British Columbia, Canada., Chan K; NanoMedicines Innovation Network, Vancouver, British Columbia, Canada; University of British Columbia (UBC), Department of Biochemistry and Molecular Biology, Vancouver, British Columbia, Canada., Witzigmann D; NanoVation Therapeutics Inc. Vancouver, British Columbia, Canada; NanoMedicines Innovation Network, Vancouver, British Columbia, Canada; University of British Columbia (UBC), Department of Biochemistry and Molecular Biology, Vancouver, British Columbia, Canada., Cullis PR; NanoVation Therapeutics Inc. Vancouver, British Columbia, Canada; NanoMedicines Innovation Network, Vancouver, British Columbia, Canada; University of British Columbia (UBC), Department of Biochemistry and Molecular Biology, Vancouver, British Columbia, Canada., Sin DD; University of British Columbia (UBC) Center for Heart Lung Innovation, St. Paul's Hospital, Vancouver, British Columbia, Canada., Lim CJ; Michael Cuccione Childhood Cancer Research Program, BC Children's Hospital Research Institute, University of British Columbia Vancouver, British Columbia, Canada. Electronic address: cjlim@mail.ubc.ca.
Jazyk: angličtina
Zdroj: European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences [Eur J Pharm Sci] 2022 Sep 01; Vol. 176, pp. 106234. Date of Electronic Publication: 2022 Jun 08.
DOI: 10.1016/j.ejps.2022.106234
Abstrakt: Introduction: Lipid nanoparticles (LNP) have been successfully used as a platform technology for delivering nucleic acids to the liver. To broaden the application of LNPs in targeting non-hepatic tissues, we developed LNP-based RNA therapies (siRNA or mRNA) for the respiratory tract. Such optimized LNP systems could offer an early treatment strategy for viral respiratory tract infections such as COVID-19.
Methods: We generated a small library of six LNP formulations with varying helper lipid compositions and characterized their hydrodynamic diameter, size distribution and cargo entrapment properties. Next, we screened these LNP formulations for particle uptake and evaluated their potential for transfecting mRNA encoding green fluorescence protein (GFP) or SARS-CoV2 nucleocapsid-GFP fusion reporter gene in a human airway epithelial cell line in vitro. Following LNP-siGFP delivery, GFP protein knockdown efficiency was assessed by flow cytometry to determine %GFP+ cells and median fluorescence intensity (MFI) for GFP. Finally, lead LNP candidates were validated in Friend leukemia virus B (FVB) male mice via intranasal delivery of an mRNA encoding luciferase, using in vivo bioluminescence imaging.
Results: Dynamic light scattering revealed that all LNP formulations contained particles with an average diameter of <100 nm and a polydispersity index of <0.2. Human airway epithelial cell lines in culture internalized LNPs with differential GFP transfection efficiencies (73-97%). The lead formulation LNP6 entrapping GFP or Nuc-GFP mRNA demonstrated the highest transfection efficiency (97%). Administration of LNP-GFP siRNA resulted in a significant reduction of GFP protein expression. For in vivo studies, intranasal delivery of LNPs containing helper lipids (DSPC, DOPC, ESM or DOPS) with luciferase mRNA showed significant increase in luminescence expression in nasal cavity and lungs by at least 10 times above baseline control.
Conclusion: LNP formulations enable the delivery of RNA payloads into human airway epithelial cells, and in the murine respiratory system; they can be delivered to nasal mucosa and lower respiratory tract via intranasal delivery. The composition of helper lipids in LNPs crucially modulates transfection efficiencies in airway epithelia, highlighting their importance in effective delivery of therapeutic products for airways diseases.
(Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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