Vasopressin V2R-targeting peptide carrier mediates siRNA delivery into collecting duct cells.

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 A...

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Main Authors: Hyun Jun Jung, Jung-Suk Lim, Hyo-Jung Choi, Mi Suk Lee, Jong-Ho Kim, Sang-Yeob Kim, Soyoun Kim, Eunjung Kim, Tae-Hwan Kwon
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2012-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3386242?pdf=render
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spelling doaj-ec113532237a49f89d7b0236bc6141282020-11-24T21:35:23ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0176e4001010.1371/journal.pone.0040010Vasopressin V2R-targeting peptide carrier mediates siRNA delivery into collecting duct cells.Hyun Jun JungJung-Suk LimHyo-Jung ChoiMi Suk LeeJong-Ho KimSang-Yeob KimSoyoun KimEunjung KimTae-Hwan KwonInternalization 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 cells; and 5) AQP2-siRNA/dDAVP-9r polyplex effectively delivered siRNA into the IMCD cells, resulting in the significant decrease of protein abundance of AQP2, but not AQP4. Therefore, for the first time to our knowledge, we demonstrated that V2R-mediated siRNA delivery could be exploited to deliver specific siRNA to regulate abnormal expression of target proteins in V2R-expressing kidney cells. The methods could be potentially used in vivo to regulate abnormal expression of proteins associated with disease conditions in the V2R-expressing kidney cells.http://europepmc.org/articles/PMC3386242?pdf=render
collection DOAJ
language English
format Article
sources DOAJ
author Hyun Jun Jung
Jung-Suk Lim
Hyo-Jung Choi
Mi Suk Lee
Jong-Ho Kim
Sang-Yeob Kim
Soyoun Kim
Eunjung Kim
Tae-Hwan Kwon
spellingShingle Hyun Jun Jung
Jung-Suk Lim
Hyo-Jung Choi
Mi Suk Lee
Jong-Ho Kim
Sang-Yeob Kim
Soyoun Kim
Eunjung Kim
Tae-Hwan Kwon
Vasopressin V2R-targeting peptide carrier mediates siRNA delivery into collecting duct cells.
PLoS ONE
author_facet Hyun Jun Jung
Jung-Suk Lim
Hyo-Jung Choi
Mi Suk Lee
Jong-Ho Kim
Sang-Yeob Kim
Soyoun Kim
Eunjung Kim
Tae-Hwan Kwon
author_sort Hyun Jun Jung
title Vasopressin V2R-targeting peptide carrier mediates siRNA delivery into collecting duct cells.
title_short Vasopressin V2R-targeting peptide carrier mediates siRNA delivery into collecting duct cells.
title_full Vasopressin V2R-targeting peptide carrier mediates siRNA delivery into collecting duct cells.
title_fullStr Vasopressin V2R-targeting peptide carrier mediates siRNA delivery into collecting duct cells.
title_full_unstemmed Vasopressin V2R-targeting peptide carrier mediates siRNA delivery into collecting duct cells.
title_sort vasopressin v2r-targeting peptide carrier mediates sirna delivery into collecting duct cells.
publisher Public Library of Science (PLoS)
series PLoS ONE
issn 1932-6203
publishDate 2012-01-01
description 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 cells; and 5) AQP2-siRNA/dDAVP-9r polyplex effectively delivered siRNA into the IMCD cells, resulting in the significant decrease of protein abundance of AQP2, but not AQP4. Therefore, for the first time to our knowledge, we demonstrated that V2R-mediated siRNA delivery could be exploited to deliver specific siRNA to regulate abnormal expression of target proteins in V2R-expressing kidney cells. The methods could be potentially used in vivo to regulate abnormal expression of proteins associated with disease conditions in the V2R-expressing kidney cells.
url http://europepmc.org/articles/PMC3386242?pdf=render
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