Extracellular ATP is involved in dsRNA-induced MUC5AC production via P2Y2R in human airway epithelium

Yutaka Shishikura, Akira Koarai, Hiroyuki Aizawa, Mutsuo Yamaya, Hisatoshi Sugiura, Mika Watanabe, Yuichiro Hashimoto, Tadahisa Numakura, Tomonori Makiguti, Kyoko Abe, Mituhiro Yamada, Toshiaki Kikuchi, Yasushi Hoshikawa, Yoshinori Okada, Masakazu Ichinose

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: In response to tissue damage or inflammation, adenosine-5'-triphosphate (ATP) is released into the extracellular compartment and has been demonstrated to augment inflammation via purinergic P2 receptors (P2Rs). Recently, ATP has been shown to be increased in the airways of COPD patients. In the present study, we examined the possible involvement of extracellular ATP in airway mucus hypersecretion during viral-induced COPD exacerbations. Methods: The involvement of extracellular ATP in the release of a major airway mucin, MUC5AC, and its signal pathway was examined after stimulation with polyinosine-polycytidylic acid [poly(I:C)], a synthetic analog of dsRNA to mimic viral infection, and rhinovirus (RV) infection in NCI-H292 cells and differentiated airway epithelial cells from COPD patients. Results: Treatment with poly(I:C) significantly increased the amount of extracellular ATP and induced MUC5AC release in NCI-H292 cells. Pre-treatment with a pannexin channel inhibitor, carbenoxolone (CBX), reduced the amount of extracellular ATP and suppressed MUC5AC release from poly(I:C)-treated cells. Pre-treatment with the P2R antagonist suramin significantly reduced the expression and release of MUC5AC. The inhibitory effects of CBX and suramin on the release of ATP and/or MUC5AC were replicated with RV infection. Pre-treatment with suramin also significantly reduced the expression and amount of extracellular EGFR ligands and the phosphorylation of EGFR and ERK in poly(I:C)-treated cells. In addition, pre-treatment with a P2Y2 receptor siRNA significantly suppressed the poly(I:C)-potentiated EGFR ligands and MUC5AC release. After poly(I:C) stimulation, the expression of MUC5AC in the differentiated cells from COPD patients was significantly higher than those from healthy subjects and the values of MUC5AC expression were inversely related with forced expiratory volume in 1 s (FEV1) % predicted. The inhibitory effects of CBX and suramin on poly(I:C)-potentiated MUC5AC expression were confirmed in differentiated airway epithelium from COPD patients. Conclusions: These results demonstrate that dsRNA induces the release of ATP via pannexin channel and that the extracellular ATP is involved in the expression and release of MUC5AC, mainly via P2Y2R, in an autocrine manner. Modulation of this pathway could be a therapeutic target for viral-induced mucus hypersecretion in COPD exacerbations.

Original languageEnglish
Article number121
JournalRespiratory Research
Volume17
Issue number1
DOIs
Publication statusPublished - 27-09-2016

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Poly C
Epithelium
Adenosine Triphosphate
Chronic Obstructive Pulmonary Disease
Suramin
Carbenoxolone
Rhinovirus
Mucus
Therapeutics
Purinergic P2Y2 Receptors
Ligands
Inflammation
Purinergic P2 Receptors
Forced Expiratory Volume
Mucins
Virus Diseases
Infection
Small Interfering RNA
Signal Transduction
Healthy Volunteers

All Science Journal Classification (ASJC) codes

  • Pulmonary and Respiratory Medicine

Cite this

Shishikura, Y., Koarai, A., Aizawa, H., Yamaya, M., Sugiura, H., Watanabe, M., ... Ichinose, M. (2016). Extracellular ATP is involved in dsRNA-induced MUC5AC production via P2Y2R in human airway epithelium. Respiratory Research, 17(1), [121]. https://doi.org/10.1186/s12931-016-0438-0
Shishikura, Yutaka ; Koarai, Akira ; Aizawa, Hiroyuki ; Yamaya, Mutsuo ; Sugiura, Hisatoshi ; Watanabe, Mika ; Hashimoto, Yuichiro ; Numakura, Tadahisa ; Makiguti, Tomonori ; Abe, Kyoko ; Yamada, Mituhiro ; Kikuchi, Toshiaki ; Hoshikawa, Yasushi ; Okada, Yoshinori ; Ichinose, Masakazu. / Extracellular ATP is involved in dsRNA-induced MUC5AC production via P2Y2R in human airway epithelium. In: Respiratory Research. 2016 ; Vol. 17, No. 1.
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abstract = "Background: In response to tissue damage or inflammation, adenosine-5'-triphosphate (ATP) is released into the extracellular compartment and has been demonstrated to augment inflammation via purinergic P2 receptors (P2Rs). Recently, ATP has been shown to be increased in the airways of COPD patients. In the present study, we examined the possible involvement of extracellular ATP in airway mucus hypersecretion during viral-induced COPD exacerbations. Methods: The involvement of extracellular ATP in the release of a major airway mucin, MUC5AC, and its signal pathway was examined after stimulation with polyinosine-polycytidylic acid [poly(I:C)], a synthetic analog of dsRNA to mimic viral infection, and rhinovirus (RV) infection in NCI-H292 cells and differentiated airway epithelial cells from COPD patients. Results: Treatment with poly(I:C) significantly increased the amount of extracellular ATP and induced MUC5AC release in NCI-H292 cells. Pre-treatment with a pannexin channel inhibitor, carbenoxolone (CBX), reduced the amount of extracellular ATP and suppressed MUC5AC release from poly(I:C)-treated cells. Pre-treatment with the P2R antagonist suramin significantly reduced the expression and release of MUC5AC. The inhibitory effects of CBX and suramin on the release of ATP and/or MUC5AC were replicated with RV infection. Pre-treatment with suramin also significantly reduced the expression and amount of extracellular EGFR ligands and the phosphorylation of EGFR and ERK in poly(I:C)-treated cells. In addition, pre-treatment with a P2Y2 receptor siRNA significantly suppressed the poly(I:C)-potentiated EGFR ligands and MUC5AC release. After poly(I:C) stimulation, the expression of MUC5AC in the differentiated cells from COPD patients was significantly higher than those from healthy subjects and the values of MUC5AC expression were inversely related with forced expiratory volume in 1 s (FEV1) {\%} predicted. The inhibitory effects of CBX and suramin on poly(I:C)-potentiated MUC5AC expression were confirmed in differentiated airway epithelium from COPD patients. Conclusions: These results demonstrate that dsRNA induces the release of ATP via pannexin channel and that the extracellular ATP is involved in the expression and release of MUC5AC, mainly via P2Y2R, in an autocrine manner. Modulation of this pathway could be a therapeutic target for viral-induced mucus hypersecretion in COPD exacerbations.",
author = "Yutaka Shishikura and Akira Koarai and Hiroyuki Aizawa and Mutsuo Yamaya and Hisatoshi Sugiura and Mika Watanabe and Yuichiro Hashimoto and Tadahisa Numakura and Tomonori Makiguti and Kyoko Abe and Mituhiro Yamada and Toshiaki Kikuchi and Yasushi Hoshikawa and Yoshinori Okada and Masakazu Ichinose",
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Shishikura, Y, Koarai, A, Aizawa, H, Yamaya, M, Sugiura, H, Watanabe, M, Hashimoto, Y, Numakura, T, Makiguti, T, Abe, K, Yamada, M, Kikuchi, T, Hoshikawa, Y, Okada, Y & Ichinose, M 2016, 'Extracellular ATP is involved in dsRNA-induced MUC5AC production via P2Y2R in human airway epithelium', Respiratory Research, vol. 17, no. 1, 121. https://doi.org/10.1186/s12931-016-0438-0

Extracellular ATP is involved in dsRNA-induced MUC5AC production via P2Y2R in human airway epithelium. / Shishikura, Yutaka; Koarai, Akira; Aizawa, Hiroyuki; Yamaya, Mutsuo; Sugiura, Hisatoshi; Watanabe, Mika; Hashimoto, Yuichiro; Numakura, Tadahisa; Makiguti, Tomonori; Abe, Kyoko; Yamada, Mituhiro; Kikuchi, Toshiaki; Hoshikawa, Yasushi; Okada, Yoshinori; Ichinose, Masakazu.

In: Respiratory Research, Vol. 17, No. 1, 121, 27.09.2016.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Extracellular ATP is involved in dsRNA-induced MUC5AC production via P2Y2R in human airway epithelium

AU - Shishikura, Yutaka

AU - Koarai, Akira

AU - Aizawa, Hiroyuki

AU - Yamaya, Mutsuo

AU - Sugiura, Hisatoshi

AU - Watanabe, Mika

AU - Hashimoto, Yuichiro

AU - Numakura, Tadahisa

AU - Makiguti, Tomonori

AU - Abe, Kyoko

AU - Yamada, Mituhiro

AU - Kikuchi, Toshiaki

AU - Hoshikawa, Yasushi

AU - Okada, Yoshinori

AU - Ichinose, Masakazu

PY - 2016/9/27

Y1 - 2016/9/27

N2 - Background: In response to tissue damage or inflammation, adenosine-5'-triphosphate (ATP) is released into the extracellular compartment and has been demonstrated to augment inflammation via purinergic P2 receptors (P2Rs). Recently, ATP has been shown to be increased in the airways of COPD patients. In the present study, we examined the possible involvement of extracellular ATP in airway mucus hypersecretion during viral-induced COPD exacerbations. Methods: The involvement of extracellular ATP in the release of a major airway mucin, MUC5AC, and its signal pathway was examined after stimulation with polyinosine-polycytidylic acid [poly(I:C)], a synthetic analog of dsRNA to mimic viral infection, and rhinovirus (RV) infection in NCI-H292 cells and differentiated airway epithelial cells from COPD patients. Results: Treatment with poly(I:C) significantly increased the amount of extracellular ATP and induced MUC5AC release in NCI-H292 cells. Pre-treatment with a pannexin channel inhibitor, carbenoxolone (CBX), reduced the amount of extracellular ATP and suppressed MUC5AC release from poly(I:C)-treated cells. Pre-treatment with the P2R antagonist suramin significantly reduced the expression and release of MUC5AC. The inhibitory effects of CBX and suramin on the release of ATP and/or MUC5AC were replicated with RV infection. Pre-treatment with suramin also significantly reduced the expression and amount of extracellular EGFR ligands and the phosphorylation of EGFR and ERK in poly(I:C)-treated cells. In addition, pre-treatment with a P2Y2 receptor siRNA significantly suppressed the poly(I:C)-potentiated EGFR ligands and MUC5AC release. After poly(I:C) stimulation, the expression of MUC5AC in the differentiated cells from COPD patients was significantly higher than those from healthy subjects and the values of MUC5AC expression were inversely related with forced expiratory volume in 1 s (FEV1) % predicted. The inhibitory effects of CBX and suramin on poly(I:C)-potentiated MUC5AC expression were confirmed in differentiated airway epithelium from COPD patients. Conclusions: These results demonstrate that dsRNA induces the release of ATP via pannexin channel and that the extracellular ATP is involved in the expression and release of MUC5AC, mainly via P2Y2R, in an autocrine manner. Modulation of this pathway could be a therapeutic target for viral-induced mucus hypersecretion in COPD exacerbations.

AB - Background: In response to tissue damage or inflammation, adenosine-5'-triphosphate (ATP) is released into the extracellular compartment and has been demonstrated to augment inflammation via purinergic P2 receptors (P2Rs). Recently, ATP has been shown to be increased in the airways of COPD patients. In the present study, we examined the possible involvement of extracellular ATP in airway mucus hypersecretion during viral-induced COPD exacerbations. Methods: The involvement of extracellular ATP in the release of a major airway mucin, MUC5AC, and its signal pathway was examined after stimulation with polyinosine-polycytidylic acid [poly(I:C)], a synthetic analog of dsRNA to mimic viral infection, and rhinovirus (RV) infection in NCI-H292 cells and differentiated airway epithelial cells from COPD patients. Results: Treatment with poly(I:C) significantly increased the amount of extracellular ATP and induced MUC5AC release in NCI-H292 cells. Pre-treatment with a pannexin channel inhibitor, carbenoxolone (CBX), reduced the amount of extracellular ATP and suppressed MUC5AC release from poly(I:C)-treated cells. Pre-treatment with the P2R antagonist suramin significantly reduced the expression and release of MUC5AC. The inhibitory effects of CBX and suramin on the release of ATP and/or MUC5AC were replicated with RV infection. Pre-treatment with suramin also significantly reduced the expression and amount of extracellular EGFR ligands and the phosphorylation of EGFR and ERK in poly(I:C)-treated cells. In addition, pre-treatment with a P2Y2 receptor siRNA significantly suppressed the poly(I:C)-potentiated EGFR ligands and MUC5AC release. After poly(I:C) stimulation, the expression of MUC5AC in the differentiated cells from COPD patients was significantly higher than those from healthy subjects and the values of MUC5AC expression were inversely related with forced expiratory volume in 1 s (FEV1) % predicted. The inhibitory effects of CBX and suramin on poly(I:C)-potentiated MUC5AC expression were confirmed in differentiated airway epithelium from COPD patients. Conclusions: These results demonstrate that dsRNA induces the release of ATP via pannexin channel and that the extracellular ATP is involved in the expression and release of MUC5AC, mainly via P2Y2R, in an autocrine manner. Modulation of this pathway could be a therapeutic target for viral-induced mucus hypersecretion in COPD exacerbations.

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