title: Uptake, efficacy, and systemic distribution of naked, inhaled short interfering RNA (siRNA) and locked nucleic acid (LNA) antisense.
abstract: Antisense oligonucleotides (ASOs) and small interfering RNA (siRNA) promise specific correction of disease-causing gene expression. Therapeutic implementation, however, has been forestalled by poor delivery to the appropriate tissue, cell type, and subcellular compartment. Topical administration is considered to circumvent these issues. The availability of inhalation devices and unmet medical need in lung disease has focused efforts in this tissue. We report the development of a novel cell sorting method for quantitative, cell type-specific analysis of siRNA, and locked nucleic acid (LNA) ASO uptake and efficacy after intratracheal (i.t.) administration in mice. Through fluorescent dye labeling, we compare the utility of this approach to whole animal and whole tissue analysis, and examine the extent of tissue distribution. We detail rapid systemic access and renal clearance for both therapeutic classes and lack of efficacy at the protein level in lung macrophages, epithelia, or other cell types. We nevertheless observe efficient redirection of i.t. administered phosphorothioate (PS) LNA ASO to the liver and kidney leading to targeted gene knockdown. These data suggest delivery remains a key obstacle to topically administered, naked oligonucleotide efficacy in the lung and introduce inhalation as a potentially viable alternative to injection for antisense administration to the liver and kidneys.
authors.affiliation: Biotherapeutics, Pfizer Global Research and Development, Sandwich, UK. s.moschos@wmin.ac.uk
authors.forename: Sterghios A
authors.initials: SA
authors.lastname: Moschos
authors.forename: Manfred
authors.initials: M
authors.lastname: Frick
authors.forename: Bruce
authors.initials: B
authors.lastname: Taylor
authors.forename: Paul
authors.initials: P
authors.lastname: Turnpenny
authors.forename: Helen
authors.initials: H
authors.lastname: Graves
authors.forename: Karen G
authors.initials: KG
authors.lastname: Spink
authors.forename: Kevin
authors.initials: K
authors.lastname: Brady
authors.forename: David
authors.initials: D
authors.lastname: Lamb
authors.forename: David
authors.initials: D
authors.lastname: Collins
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Rockel
authors.forename: Markus
authors.initials: M
authors.lastname: Weber
authors.forename: Ovadia
authors.initials: O
authors.lastname: Lazari
authors.forename: Luis
authors.initials: L
authors.lastname: Perez-Tosar
authors.forename: Sally A
authors.initials: SA
authors.lastname: Fancy
authors.forename: Chris
authors.initials: C
authors.lastname: Lapthorn
authors.forename: Martin X
authors.initials: MX
authors.lastname: Green
authors.forename: Steve
authors.initials: S
authors.lastname: Evans
authors.forename: Matthew
authors.initials: M
authors.lastname: Selby
authors.forename: Gareth
authors.initials: G
authors.lastname: Jones
authors.forename: Lyn
authors.initials: L
authors.lastname: Jones
authors.forename: Sarah
authors.initials: S
authors.lastname: Kearney
authors.forename: Houria
authors.initials: H
authors.lastname: Mechiche
authors.forename: Diana
authors.initials: D
authors.lastname: Gikunju
authors.forename: Romesh
authors.initials: R
authors.lastname: Subramanian
authors.forename: Eugen
authors.initials: E
authors.lastname: Uhlmann
authors.forename: Marion
authors.initials: M
authors.lastname: Jurk
authors.forename: Jörg
authors.initials: J
authors.lastname: Vollmer
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Ciaramella
authors.forename: Michael
authors.initials: M
authors.lastname: Yeadon
mesh_terms: D000280:Administration, Inhalation; D000818:Animals; D001055:Apolipoproteins B / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D020868:Gene Silencing; D018390:Gene Targeting; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008156:Luciferases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009841:Oligonucleotides / Q000008:administration & dosage / Q000493:pharmacokinetics*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000493:pharmacokinetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D014018:Tissue Distribution
chemical_list: D001055:Apolipoproteins B; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; C477371:locked nucleic acid; D049452:Green Fluorescent Proteins; D008156:Luciferases