原创力文档vasopressin v2r-targeting peptide carrier mediates sirna delivery into collecting duct cells后叶加压素v2r-targeting肽载体介导核交付为集合管细胞.pdfVIP
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vasopressin v2r-targeting peptide carrier mediates sirna delivery into collecting duct cells后叶加压素v2r-targeting肽载体介导核交付为集合管细胞
Vasopressin V2R-Targeting Peptide Carrier Mediates
siRNA Delivery into Collecting Duct Cells
1 1 1 1 1,2 1
Hyun Jun Jung , Jung-Suk Lim , Hyo-Jung Choi , Mi Suk Lee , Jong-Ho Kim , Sang-Yeob Kim ,
1 1 1
Soyoun Kim , Eunjung Kim , Tae-Hwan Kwon *
1 Department of Biochemistry and Cell Biology, School of Medicine, Kyungpook National University, Taegu Korea, 2 Department of Pharmaceutical Science, College of
Pharmacy, Kyung Hee University, Seoul, Korea
Abstract
Internalization of receptor proteins after interacting with specific ligands has been proposed to facilitate siRNA delivery into
the target cells via receptor-mediated siRNA transduction. In this study, we demonstrated a novel method of vasopressin V2
receptor (V2R)-mediated siRNA delivery against AQP2 in primary cultured inner medullary collecting duct (IMCD) cells of rat
kidney. We synthesized the dDAVP conjugated with nine D-arginines (dDAVP-9r) as a peptide carrier for siRNA delivery. The
structure of synthetic peptide carrier showed two regions (i.e., ligand domain to V2R (dDAVP) and siRNA carrying domain
(nine D-arginine)) bisected with a spacer of four glycines. The results revealed that 1) synthesized dDAVP-9r peptides
formed a stable polyplex with siRNA; 2) siRNA/dDAVP-9r polyplex could bind to the V2R of IMCD cells and induced AQP2
phosphorylation (Ser 256); 3) siRNA/dDAVP-9r polyplex was stable in response to the wide range of different osmolalities,
pH levels, or to the RNases; 4) fluorescein-labeled siRNA was delivered into V2R-expressing MDCK and LLC-PK1 cells by
siRNA/dDAVP-9r polyplex, but not into the V2R-negative Cos-7 cel
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