Repression of Connexin26 hemichannel activity protects the barrier function of respiratory airway epithelial cells against LPS-induced alteration

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Authors

  • Tina Lehrich
  • Anne Dierks
  • Masina Plenge
  • Helena Obernolte
  • Klaudia Grieger
  • Katherina Sewald
  • Frederic Rodriguez
  • Lucie Malet
  • Peter Braubach
  • Florence Bedos-Belval
  • Anaclet Ngezahayo

External Research Organisations

  • Fraunhofer Institute for Toxicology and Experimental Medicine (ITEM)
  • Hannover Medical School (MHH)
  • Centre national de la recherche scientifique (CNRS)
  • Center for Systems Neuroscience Hannover (ZSN)
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Details

Original languageEnglish
Article number226
JournalCell communication and signaling
Volume23
Issue number1
Publication statusPublished - 16 May 2025

Abstract

In respiratory airway epithelial cells, lipopolysaccharide (LPS) treatment induced an enhancement of connexin 26 (Cx26) hemichannel activity shown by dye uptake experiments after siRNA-mediated knock-down of Cx26. This effect was already observed at infection relevant concentrations (≤ 10 ng/mL LPS) and involved tumor necrosis factor alpha (TNF-α)- and Ca2+-dependent signaling. High concentrations (1 µg/mL LPS) reduced the transepithelial electrical resistance (TEER) of Calu-3 cells by 35% within an application time of 3 h followed by a recovery. Parallel to barrier alteration, a reduced tight junction organization rate (TiJOR) of claudin-4 (CLDN4) by 75% was observed within an application time of 3 h. After TEER recovery, CLDN4 TiJOR stayed reduced. Low concentrations (10 ng/mL LPS) required three times repeated application for barrier reduction and CLDN4 TiJOR reduction by 30%. The small molecule CVB4-57, newly published as a potential inhibitor of Cx26 hemichannels, mitigated the effects of LPS on the epithelial barrier function. Molecular docking studies revealed a potential interaction between CVB4-57 and Cx26 thereby reducing its hemichannel activity. We conclude that LPS-related enhancement of Cx26 hemichannel activity acts like a “molecular scar” that weakens the lung epithelium, which could be attenuated by agents targeting Cx26 hemichannels.

Keywords

    Airway epithelium, Barrier function, Calu-3 cells, Connexin channels; lipopolysaccharide, Cytokine, PCLS, Primary cells

ASJC Scopus subject areas

Cite this

Repression of Connexin26 hemichannel activity protects the barrier function of respiratory airway epithelial cells against LPS-induced alteration. / Lehrich, Tina; Dierks, Anne; Plenge, Masina et al.
In: Cell communication and signaling, Vol. 23, No. 1, 226, 16.05.2025.

Research output: Contribution to journalArticleResearchpeer review

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title = "Repression of Connexin26 hemichannel activity protects the barrier function of respiratory airway epithelial cells against LPS-induced alteration",
abstract = "In respiratory airway epithelial cells, lipopolysaccharide (LPS) treatment induced an enhancement of connexin 26 (Cx26) hemichannel activity shown by dye uptake experiments after siRNA-mediated knock-down of Cx26. This effect was already observed at infection relevant concentrations (≤ 10 ng/mL LPS) and involved tumor necrosis factor alpha (TNF-α)- and Ca2+-dependent signaling. High concentrations (1 µg/mL LPS) reduced the transepithelial electrical resistance (TEER) of Calu-3 cells by 35% within an application time of 3 h followed by a recovery. Parallel to barrier alteration, a reduced tight junction organization rate (TiJOR) of claudin-4 (CLDN4) by 75% was observed within an application time of 3 h. After TEER recovery, CLDN4 TiJOR stayed reduced. Low concentrations (10 ng/mL LPS) required three times repeated application for barrier reduction and CLDN4 TiJOR reduction by 30%. The small molecule CVB4-57, newly published as a potential inhibitor of Cx26 hemichannels, mitigated the effects of LPS on the epithelial barrier function. Molecular docking studies revealed a potential interaction between CVB4-57 and Cx26 thereby reducing its hemichannel activity. We conclude that LPS-related enhancement of Cx26 hemichannel activity acts like a “molecular scar” that weakens the lung epithelium, which could be attenuated by agents targeting Cx26 hemichannels.",
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Download

TY - JOUR

T1 - Repression of Connexin26 hemichannel activity protects the barrier function of respiratory airway epithelial cells against LPS-induced alteration

AU - Lehrich, Tina

AU - Dierks, Anne

AU - Plenge, Masina

AU - Obernolte, Helena

AU - Grieger, Klaudia

AU - Sewald, Katherina

AU - Rodriguez, Frederic

AU - Malet, Lucie

AU - Braubach, Peter

AU - Bedos-Belval, Florence

AU - Ngezahayo, Anaclet

N1 - Publisher Copyright: © The Author(s) 2025.

PY - 2025/5/16

Y1 - 2025/5/16

N2 - In respiratory airway epithelial cells, lipopolysaccharide (LPS) treatment induced an enhancement of connexin 26 (Cx26) hemichannel activity shown by dye uptake experiments after siRNA-mediated knock-down of Cx26. This effect was already observed at infection relevant concentrations (≤ 10 ng/mL LPS) and involved tumor necrosis factor alpha (TNF-α)- and Ca2+-dependent signaling. High concentrations (1 µg/mL LPS) reduced the transepithelial electrical resistance (TEER) of Calu-3 cells by 35% within an application time of 3 h followed by a recovery. Parallel to barrier alteration, a reduced tight junction organization rate (TiJOR) of claudin-4 (CLDN4) by 75% was observed within an application time of 3 h. After TEER recovery, CLDN4 TiJOR stayed reduced. Low concentrations (10 ng/mL LPS) required three times repeated application for barrier reduction and CLDN4 TiJOR reduction by 30%. The small molecule CVB4-57, newly published as a potential inhibitor of Cx26 hemichannels, mitigated the effects of LPS on the epithelial barrier function. Molecular docking studies revealed a potential interaction between CVB4-57 and Cx26 thereby reducing its hemichannel activity. We conclude that LPS-related enhancement of Cx26 hemichannel activity acts like a “molecular scar” that weakens the lung epithelium, which could be attenuated by agents targeting Cx26 hemichannels.

AB - In respiratory airway epithelial cells, lipopolysaccharide (LPS) treatment induced an enhancement of connexin 26 (Cx26) hemichannel activity shown by dye uptake experiments after siRNA-mediated knock-down of Cx26. This effect was already observed at infection relevant concentrations (≤ 10 ng/mL LPS) and involved tumor necrosis factor alpha (TNF-α)- and Ca2+-dependent signaling. High concentrations (1 µg/mL LPS) reduced the transepithelial electrical resistance (TEER) of Calu-3 cells by 35% within an application time of 3 h followed by a recovery. Parallel to barrier alteration, a reduced tight junction organization rate (TiJOR) of claudin-4 (CLDN4) by 75% was observed within an application time of 3 h. After TEER recovery, CLDN4 TiJOR stayed reduced. Low concentrations (10 ng/mL LPS) required three times repeated application for barrier reduction and CLDN4 TiJOR reduction by 30%. The small molecule CVB4-57, newly published as a potential inhibitor of Cx26 hemichannels, mitigated the effects of LPS on the epithelial barrier function. Molecular docking studies revealed a potential interaction between CVB4-57 and Cx26 thereby reducing its hemichannel activity. We conclude that LPS-related enhancement of Cx26 hemichannel activity acts like a “molecular scar” that weakens the lung epithelium, which could be attenuated by agents targeting Cx26 hemichannels.

KW - Airway epithelium

KW - Barrier function

KW - Calu-3 cells

KW - Connexin channels; lipopolysaccharide

KW - Cytokine

KW - PCLS

KW - Primary cells

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U2 - 10.1186/s12964-025-02228-6

DO - 10.1186/s12964-025-02228-6

M3 - Article

C2 - 40380298

AN - SCOPUS:105005459837

VL - 23

JO - Cell communication and signaling

JF - Cell communication and signaling

SN - 1478-811X

IS - 1

M1 - 226

ER -

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