In vivo delivery of small interfering RNA targeting brain capillary endothelial cells.
Brain capillary endothelial cells (BCECs) play an important role in blood-brain barrier (BBB) functions and pathophysiologic mechanisms in brain ischemia and inflammation. We try to suppress gene expression in BCECs by intravenous application of small interfering RNA (siRNA). After injection of large dose siRNA with hydrodynamic technique to mouse, suppression of endogenous protein and the BBB function of BCECs was investigated. The brain-to-blood transport function of organic anion transporter 3 (OAT3) that expressed in BCECs was evaluated by Brain Efflux Index method in mouse. The siRNA could be delivered to BCECs and efficiently inhibited endogenously expressed protein of BCECs. The suppression effect of siRNA to OAT3 is enough to reduce the brain-to-blood transport of OAT3 substrate, benzylpenicillin at BBB. The in vivo siRNA-silencing method with hydrodynamic technique may be useful for the study of BBB function and gene therapy targeting BCECs.
Hino
Taro
T
Department of Neurology and Neurological Science, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.
Yokota
Takanori
T
Ito
Shingo
S
Nishina
Kazutaka
K
Kang
Young-Sook
YS
Mori
Shinobu
S
Hori
Satoko
S
Kanda
Takashi
T
Terasaki
Tetsuya
T
Mizusawa
Hidehiro
H
D000818:Animals; D001812:Blood-Brain Barrier / Q000502:physiology*; D001921:Brain / Q000098:blood supply* / Q000502:physiology; D002460:Cell Line; D016503:Drug Delivery Systems / Q000379:methods; D042783:Endothelial Cells / Q000502:physiology*; D020868:Gene Silencing; D018390:Gene Targeting / Q000379:methods*; D006801:Humans; D007668:Kidney / Q000502:physiology; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D029362:Organic Anion Transporters, Sodium-Independent / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D000072105:Superoxide Dismutase-1
D029362:Organic Anion Transporters, Sodium-Independent; D034741:RNA, Small Interfering; C000606290:SOD1 protein, human; C114338:organic anion transport protein 3; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1