title: Optimized siRNA-PEG conjugates for extended blood circulation and reduced urine excretion in mice.
abstract: Some of the main concerns with in vivo application of naked small interfering RNA are rapid degradation and urinary excretion resulting in a short plasma half-life. In this study we investigated how conjugation of polyethylene glycol (PEG) with variable chain length affects siRNA pharmacokinetics and biodistribution. The PEG chains were conjugated to chemically stabilized siRNA at the 5' terminal end of the passenger strand using click chemistry. The siRNA conjugate remained functionally active and showed significantly prolonged circulation in the blood stream after intravenous injection. siRNA conjugated with 20kDa PEG (PEG20k-siRNA) was most persistent, approximately 50% PEG20k-siRNA remained 1h post-injection, while the uncoupled siRNA was rapidly removed >90% at 15min. In vivo fluorescent imaging of the living animal showed increased concentration of siRNA in peripheral tissue and delayed urine excretion when coupled to PEG 20k. Biodistribution studies by northern blotting revealed equal distribution of conjugated siRNA in liver, kidney, spleen and lung without significant degradation 24 h post-injection. Our study demonstrates that PEG conjugated siRNA can be applied as a delivery system to improve siRNA bioavailability in vivo and may potentially increase the efficiency of siRNA in therapeutic applications.
authors.affiliation: Interdisciplinary Nanoscience Center (iNANO) and Department of Molecular Biology and Genetics, C.F. Møllers Alle, Building 1130, 8000 Aarhus C, Aarhus University, Aarhus, Denmark.
authors.forename: Frank
authors.initials: F
authors.lastname: Iversen
authors.forename: Chuanxu
authors.initials: C
authors.lastname: Yang
authors.forename: Frederik
authors.initials: F
authors.lastname: Dagnæs-Hansen
authors.forename: David H
authors.initials: DH
authors.lastname: Schaffert
authors.forename: Jørgen
authors.initials: J
authors.lastname: Kjems
authors.forename: Shan
authors.initials: S
authors.lastname: Gao
mesh_terms: D000825:Animal Structures / Q000000981:diagnostic imaging; D000818:Animals; D001688:Biological Products / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics*; D015152:Blotting, Northern; D006207:Half-Life; D007275:Injections, Intravenous; D051379:Mice; D061848:Optical Imaging; D010949:Plasma / Q000737:chemistry; D011092:Polyethylene Glycols / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics*; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry / Q000493:pharmacokinetics*; D011859:Radiography; D014556:Urine / Q000737:chemistry
chemical_list: D001688:Biological Products; D034741:RNA, Small Interfering; D011092:Polyethylene Glycols
keywords: PEGylation; blood circulation; in vivo imaging.; siRNA; urine excretion