title: Efficient in vivo delivery of siRNA into brain capillary endothelial cells along with endogenous lipoprotein.
abstract: The brain capillary endothelial cell (BCEC) is a major functional component of the blood-brain barrier and is an underlying factor in the pathophysiology of various diseases, including brain ischemia, multiple sclerosis, and neurodegenerative disorders. We examined gene silencing in BCECs by using endogenous lipoprotein to introduce short-interfering RNA (siRNA) in vivo. A cholesterol-conjugated 21/23-mer siRNA targeting organic anion transporter 3 (OAT3) mRNA (Chol-siOAT3) was intravenously injected into mice after its incorporation into extracted endogenous lipoproteins. Chol-siOAT3 was not delivered to neurons or glia, but was successfully delivered into BCECs and resulted in a significant reduction of OAT3 mRNA levels when injected after its incorporation into high-density lipoprotein (HDL). Efficient delivery was not achieved, however, when Chol-siOAT3 was injected without any lipoproteins, or after its incorporation into low-density lipoprotein (LDL). Investigations in apolipoprotein E (ApoE)-deficient and LDL receptor (LDLR)-deficient mice revealed that the uptake of HDL-containing Chol-siOAT3 was mainly mediated by ApoE and LDLR in mice. These findings indicate that siRNA can be delivered into BCECs in vivo by using endogenous lipoprotein, which could make this strategy useful as a new gene silencing therapy for diseases involving BCECs.
authors.affiliation: Department of Neurology and Neurological Science, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.
authors.forename: Hiroya
authors.initials: H
authors.lastname: Kuwahara
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Nishina
authors.forename: Kie
authors.initials: K
authors.lastname: Yoshida
authors.forename: Tomoko
authors.initials: T
authors.lastname: Nishina
authors.forename: Mariko
authors.initials: M
authors.lastname: Yamamoto
authors.forename: Yukari
authors.initials: Y
authors.lastname: Saito
authors.forename: Wenying
authors.initials: W
authors.lastname: Piao
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yoshida
authors.forename: Hidehiro
authors.initials: H
authors.lastname: Mizusawa
authors.forename: Takanori
authors.initials: T
authors.lastname: Yokota
mesh_terms: D000818:Animals; D001057:Apolipoproteins E / Q000502:physiology; D001812:Blood-Brain Barrier; D015152:Blotting, Northern; D001921:Brain / Q000098:blood supply / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism*; D016503:Drug Delivery Systems; D024202:Electrophoretic Mobility Shift Assay; D042783:Endothelial Cells / Q000378:metabolism*; D005260:Female; D020868:Gene Silencing; D007124:Immunoenzyme Techniques; D007275:Injections, Intravenous; D008075:Lipoproteins, HDL / Q000378:metabolism*; D008077:Lipoproteins, LDL / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D029362:Organic Anion Transporters, Sodium-Independent / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics*; D011973:Receptors, LDL / Q000502:physiology
chemical_list: D001057:Apolipoproteins E; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D029362:Organic Anion Transporters, Sodium-Independent; D034741:RNA, Small Interfering; D011973:Receptors, LDL; C114338:organic anion transport protein 3; D002784:Cholesterol