title: Delivery of ENaC siRNA to epithelial cells mediated by a targeted nanocomplex: a therapeutic strategy for cystic fibrosis.
abstract: The inhibition of ENaC may have therapeutic potential in CF airways by reducing sodium hyperabsorption, restoring lung epithelial surface fluid levels, airway hydration and mucociliary function. The challenge has been to deliver siRNA to the lung with sufficient efficacy for a sustained therapeutic effect. We have developed a self-assembling nanocomplex formulation for siRNA delivery to the airways that consists of a liposome (DOTMA/DOPE; L), an epithelial targeting peptide (P) and siRNA (R). LPR formulations were assessed for their ability to silence expression of the transcript of the gene encoding the α-subunit of the sodium channel ENaC in cell lines and primary epithelial cells, in submerged cultures or grown in air-liquid interface conditions. LPRs, containing 50 nM or 100 nM siRNA, showed high levels of silencing, particularly in primary airway epithelial cells. When nebulised these nanocomplexes still retained their biophysical properties and transfection efficiencies. The silencing ability was determined at protein level by confocal microscopy and western blotting. In vivo data demonstrated that these nanoparticles had the ability to silence expression of the α-ENaC subunit gene. In conclusion, these findings show that LPRs can modulate the activity of ENaC and this approach might be promising as co-adjuvant therapy for cystic fibrosis.
authors.affiliation: Experimental and Personalised Medicine Section, UCL Great Ormond Street Institute of Child Health, London, UK.
authors.forename: Maria D I
authors.identifier: http://orcid.org/0000-0002-1875-5335
authors.initials: MDI
authors.lastname: Manunta
authors.affiliation: Experimental and Personalised Medicine Section, UCL Great Ormond Street Institute of Child Health, London, UK.
authors.forename: Aristides D
authors.initials: AD
authors.lastname: Tagalakis
authors.affiliation: Experimental and Personalised Medicine Section, UCL Great Ormond Street Institute of Child Health, London, UK.|Centre for Cellular and Molecular Physiology, University of Oxford, Oxford, UK.
authors.forename: Martin
authors.initials: M
authors.lastname: Attwood
authors.affiliation: Experimental and Personalised Medicine Section, UCL Great Ormond Street Institute of Child Health, London, UK.
authors.forename: Ahmad M
authors.initials: AM
authors.lastname: Aldossary
authors.affiliation: UCL Respiratory Centre for Inflammation and Tissue Repair, University College London, London, UK.
authors.forename: Josephine L
authors.initials: JL
authors.lastname: Barnes
authors.affiliation: Experimental and Personalised Medicine Section, UCL Great Ormond Street Institute of Child Health, London, UK.
authors.forename: Mustafa M
authors.initials: MM
authors.lastname: Munye
authors.affiliation: Experimental and Personalised Medicine Section, UCL Great Ormond Street Institute of Child Health, London, UK.|Institute of Pharmacy, Freie Universität Berlin, Berlin, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Weng
authors.affiliation: UCL Respiratory Centre for Inflammation and Tissue Repair, University College London, London, UK.
authors.forename: Robin J
authors.identifier: http://orcid.org/0000-0003-1936-4245
authors.initials: RJ
authors.lastname: McAnulty
authors.affiliation: Experimental and Personalised Medicine Section, UCL Great Ormond Street Institute of Child Health, London, UK. s.hart@ucl.ac.uk.
authors.forename: Stephen L
authors.identifier: http://orcid.org/0000-0001-8254-376X
authors.initials: SL
authors.lastname: Hart
mesh_terms: D002460:Cell Line; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000235:genetics / Q000628:therapy; D004847:Epithelial Cells / Q000378:metabolism*; D053503:Epithelial Sodium Channels / Q000235:genetics*; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D008081:Liposomes / Q000737:chemistry; D018613:Microscopy, Confocal; D053758:Nanoparticles; D010455:Peptides / Q000737:chemistry; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics*; D014161:Transduction, Genetic; D014162:Transfection
chemical_list: D053503:Epithelial Sodium Channels; D008081:Liposomes; D010455:Peptides; D034741:RNA, Small Interfering