Uptake, Efficacy, and Systemic Distribution of Naked, Inhaled Short Interfering RNA (siRNA) and Locked Nucleic Acid (LNA) Antisense
| Autor: | Romesh R. Subramanian, David J. Lamb, Sterghios Moschos, Houria Mechiche, Bruce M. Taylor, Steve Evans, Chris Lapthorn, Michael Yeadon, Kevin Brady, Luis Perez-Tosar, Marion Jurk, Manfred Frick, Sally A. Fancy, Eugen Uhlmann, David Collins, Ovadia Lazari, Lyn H. Jones, Karen G. Spink, Thomas Dino Rockel, Sarah Kearney, Paul Turnpenny, Martin X. Green, Helen Graves, Joerg Vollmer, Gareth Jones, Markus Weber, Matthew D. Selby, Giuseppe Ciaramella, Diana Gikunju |
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| Rok vydání: | 2011 |
| Předmět: |
Male
Small interfering RNA Cell Green Fluorescent Proteins Oligonucleotides Mice Transgenic Pharmacology Biology Kidney Mice Gene expression Administration Inhalation Drug Discovery medicine Genetics Distribution (pharmacology) Animals Tissue Distribution Gene Silencing Locked nucleic acid RNA Small Interfering Luciferases Molecular Biology Cells Cultured Apolipoproteins B Mice Knockout Gene knockdown Mice Inbred BALB C Oligonucleotide Cell sorting Oligonucleotides Antisense Molecular biology medicine.anatomical_structure Liver Gene Targeting Molecular Medicine Original Article |
| Zdroj: | Molecular Therapy |
| ISSN: | 1525-0016 |
| DOI: | 10.1038/mt.2011.206 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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