Specific inhibition of epithelial Na+ channels by antisense oligonucleotides for the treatment of Na+ hyperabsorption in cystic fibrosis

Publication type:
Journal article
Metadata:
Autoren
  • Katja Sobczak
  • Andrei Segal
  • Nadine Bangel-Ruland
  • Judith Semmler
  • Willy Van Driessche
  • Hermann Lindemann
  • Ralf Heermann
  • Wolf-Michael Weber
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000270043300008&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1002/jgm.1363
eISSN
1521-2254
Externe Identifier
  • Clarivate Analytics Document Solution ID: 497FQ
  • PubMed Identifier: 19533590
ISSN
1099-498X
Ausgabe der Veröffentlichung
9
Zeitschrift
JOURNAL OF GENE MEDICINE
Schlüsselwörter
  • antisense oligonucleotides
  • cystic fibrosis
  • epithelial Na+ channel
  • Na+ hyperabsorption
Paginierung
813 - 823
Datum der Veröffentlichung
2009
Status
Published
Titel
Specific inhibition of epithelial Na<SUP>+</SUP> channels by antisense oligonucleotides for the treatment of Na<SUP>+</SUP> hyperabsorption in cystic fibrosis
Sub types
  • Article
Ausgabe der Zeitschrift
11

Data source: Web of Science (Lite)

Other metadata sources:
Abstract
<jats:title>Abstract</jats:title><jats:sec><jats:title>Background</jats:title><jats:p>Cystic fibrosis (CF) respiratory epithelia are characterized by a defect Cl<jats:sup>−</jats:sup> secretion and an increased Na<jats:sup>+</jats:sup> absorption through epithelial Na<jats:sup>+</jats:sup> channels (ENaC). The present study aimed to find an effective inhibitor of human ENaC with respect to replacing amiloride therapy for CF patients. Therefore, we developed specific antisense oligonucleotides (AON) that efficiently suppress Na<jats:sup>+</jats:sup> hyperabsorption by inhibiting the expression of the α‐ENaC subunit.</jats:p></jats:sec><jats:sec><jats:title>Methods</jats:title><jats:p>We heterologously expressed ENaC in oocytes of <jats:italic>Xenopus laevis</jats:italic> for mass screening of AON. Additionally, primary cultures of human nasal epithelia were transfected with AON and were used for Ussing chamber experiments, as well as biochemical and fluorescence optical analyses.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p>Screening of several AON by co‐injection or sequential microinjection of AON and ENaC mRNA in <jats:italic>X. laevis</jats:italic> oocytes led to a sustained decrease in amiloride‐sensitive current and conductance. Using primary cultures of human nasal epithelia, we show that AON effectively suppress amiloride‐sensitive Na<jats:sup>+</jats:sup> absorption mediated by ENaC in CF and non‐CF tissues. In western blot experiments, it could be shown that the amount of ENaC protein is effectively reduced after AON transfection.</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>Our data comprise an initial step towards a preclinical test with AON to reduce Na<jats:sup>+</jats:sup> hyperabsorption in CF epithelia. Copyright © 2009 John Wiley &amp; Sons, Ltd.</jats:p></jats:sec>
Autoren
  • Katja Sobczak
  • Andrei Segal
  • Nadine Bangel‐Ruland
  • Judith Semmler
  • Willy Van Driessche
  • Hermann Lindemann
  • Ralf Heermann
  • Wolf‐Michael Weber
DOI
10.1002/jgm.1363
eISSN
1521-2254
ISSN
1099-498X
Ausgabe der Veröffentlichung
9
Zeitschrift
The Journal of Gene Medicine
Sprache
en
Online publication date
2009
Paginierung
813 - 823
Datum der Veröffentlichung
2009
Status
Published
Herausgeber
Wiley
Herausgeber URL
http://dx.doi.org/10.1002/jgm.1363
Datum der Datenerfassung
2023
Titel
Specific inhibition of epithelial Na<sup>+</sup> channels by antisense oligonucleotides for the treatment of Na<sup>+</sup> hyperabsorption in cystic fibrosis
Ausgabe der Zeitschrift
11

Data source: Crossref

Abstract
<h4>Background</h4>Cystic fibrosis (CF) respiratory epithelia are characterized by a defect Cl(-) secretion and an increased Na(+) absorption through epithelial Na(+) channels (ENaC). The present study aimed to find an effective inhibitor of human ENaC with respect to replacing amiloride therapy for CF patients. Therefore, we developed specific antisense oligonucleotides (AON) that efficiently suppress Na(+) hyperabsorption by inhibiting the expression of the alpha-ENaC subunit.<h4>Methods</h4>We heterologously expressed ENaC in oocytes of Xenopus laevis for mass screening of AON. Additionally, primary cultures of human nasal epithelia were transfected with AON and were used for Ussing chamber experiments, as well as biochemical and fluorescence optical analyses.<h4>Results</h4>Screening of several AON by co-injection or sequential microinjection of AON and ENaC mRNA in X. laevis oocytes led to a sustained decrease in amiloride-sensitive current and conductance. Using primary cultures of human nasal epithelia, we show that AON effectively suppress amiloride-sensitive Na(+) absorption mediated by ENaC in CF and non-CF tissues. In western blot experiments, it could be shown that the amount of ENaC protein is effectively reduced after AON transfection.<h4>Conclusions</h4>Our data comprise an initial step towards a preclinical test with AON to reduce Na(+) hyperabsorption in CF epithelia.
Addresses
  • Institute of Animal Physiology, Westphalian Wilhelms-University Muenster, Muenster, Germany.
Autoren
  • Katja Sobczak
  • Andrei Segal
  • Nadine Bangel-Ruland
  • Judith Semmler
  • Willy Van Driessche
  • Hermann Lindemann
  • Ralf Heermann
  • Wolf-Michael Weber
DOI
10.1002/jgm.1363
eISSN
1521-2254
Externe Identifier
  • PubMed Identifier: 19533590
Open access
false
ISSN
1099-498X
Ausgabe der Veröffentlichung
9
Zeitschrift
The journal of gene medicine
Schlüsselwörter
  • Nasal Mucosa
  • Animals
  • Xenopus laevis
  • Humans
  • Cystic Fibrosis
  • Sodium
  • Oligonucleotides, Antisense
  • RNA, Messenger
  • DNA Primers
  • Microscopy, Fluorescence
  • Microelectrodes
  • Base Sequence
  • Epithelial Sodium Channels
  • Epithelial Sodium Channel Blockers
Sprache
eng
Medium
Print
Paginierung
813 - 823
Datum der Veröffentlichung
2009
Status
Published
Datum der Datenerfassung
2009
Titel
Specific inhibition of epithelial Na+ channels by antisense oligonucleotides for the treatment of Na+ hyperabsorption in cystic fibrosis.
Sub types
  • Research Support, Non-U.S. Gov't
  • Journal Article
Ausgabe der Zeitschrift
11

Data source: Europe PubMed Central

Abstract
BACKGROUND: Cystic fibrosis (CF) respiratory epithelia are characterized by a defect Cl(-) secretion and an increased Na(+) absorption through epithelial Na(+) channels (ENaC). The present study aimed to find an effective inhibitor of human ENaC with respect to replacing amiloride therapy for CF patients. Therefore, we developed specific antisense oligonucleotides (AON) that efficiently suppress Na(+) hyperabsorption by inhibiting the expression of the alpha-ENaC subunit. METHODS: We heterologously expressed ENaC in oocytes of Xenopus laevis for mass screening of AON. Additionally, primary cultures of human nasal epithelia were transfected with AON and were used for Ussing chamber experiments, as well as biochemical and fluorescence optical analyses. RESULTS: Screening of several AON by co-injection or sequential microinjection of AON and ENaC mRNA in X. laevis oocytes led to a sustained decrease in amiloride-sensitive current and conductance. Using primary cultures of human nasal epithelia, we show that AON effectively suppress amiloride-sensitive Na(+) absorption mediated by ENaC in CF and non-CF tissues. In western blot experiments, it could be shown that the amount of ENaC protein is effectively reduced after AON transfection. CONCLUSIONS: Our data comprise an initial step towards a preclinical test with AON to reduce Na(+) hyperabsorption in CF epithelia.
Autoren
  • Katja Sobczak
  • Andrei Segal
  • Nadine Bangel-Ruland
  • Judith Semmler
  • Willy Van Driessche
  • Hermann Lindemann
  • Ralf Heermann
  • Wolf-Michael Weber
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/19533590
DOI
10.1002/jgm.1363
eISSN
1521-2254
Ausgabe der Veröffentlichung
9
Zeitschrift
J Gene Med
Schlüsselwörter
  • Animals
  • Base Sequence
  • Cystic Fibrosis
  • DNA Primers
  • Epithelial Sodium Channel Blockers
  • Epithelial Sodium Channels
  • Humans
  • Microelectrodes
  • Microscopy, Fluorescence
  • Nasal Mucosa
  • Oligonucleotides, Antisense
  • RNA, Messenger
  • Sodium
  • Xenopus laevis
Sprache
eng
Country
England
Paginierung
813 - 823
Datum der Veröffentlichung
2009
Status
Published
Datum, an dem der Datensatz öffentlich gemacht wurde
2009
Titel
Specific inhibition of epithelial Na+ channels by antisense oligonucleotides for the treatment of Na+ hyperabsorption in cystic fibrosis.
Sub types
  • Journal Article
  • Research Support, Non-U.S. Gov't
Ausgabe der Zeitschrift
11

Data source: PubMed

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