Transient receptor potential canonical-3 channel-dependent fibroblast regulation in atrial fibrillation

Masahide Harada, Xiaobin Luo, Xiao Yan Qi, Artavazd Tadevosyan, Ange Maguy, Balazs Ordog, Jonathan Ledoux, Takeshi Kato, Patrice Naud, Niels Voigt, Yanfen Shi, Kaichiro Kamiya, Toyoaki Murohara, Itsuo Kodama, Jean Claude Tardif, Ulrich Schotten, David R. Van Wagoner, Dobromir Dobrev, Stanley Nattel

Research output: Contribution to journalArticle

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Abstract

BACKGROUND-: Fibroblast proliferation and differentiation are central in atrial fibrillation (AF)-promoting remodeling. Here, we investigated fibroblast regulation by Ca-permeable transient receptor potential canonical-3 (TRPC3) channels. METHODS AND RESULTS-: Freshly isolated rat cardiac fibroblasts abundantly expressed TRPC3 and had appreciable nonselective cation currents (INSC) sensitive to a selective TPRC3 channel blocker, pyrazole-3 (3 μmol/L). Pyrazole-3 suppressed angiotensin II-induced Ca influx, proliferation, and α-smooth muscle actin protein expression in fibroblasts. Ca removal and TRPC3 blockade suppressed extracellular signal-regulated kinase phosphorylation, and extracellular signal-regulated kinase phosphorylation inhibition reduced fibroblast proliferation. TRPC3 expression was upregulated in atria from AF patients, goats with electrically maintained AF, and dogs with tachypacing-induced heart failure. TRPC3 knockdown (based on short hairpin RNA [shRNA]) decreased canine atrial fibroblast proliferation. In left atrial fibroblasts freshly isolated from dogs kept in AF for 1 week by atrial tachypacing, TRPC3 protein expression, currents, extracellular signal-regulated kinase phosphorylation, and extracellular matrix gene expression were all significantly increased. In cultured left atrial fibroblasts from AF dogs, proliferation rates, α-smooth muscle actin expression, and extracellular signal-regulated kinase phosphorylation were increased and were suppressed by pyrazole-3. MicroRNA-26 was downregulated in canine AF atria; experimental microRNA-26 knockdown reproduced AF-induced TRPC3 upregulation and fibroblast activation. MicroRNA-26 has NFAT (nuclear factor of activated T cells) binding sites in the 5′ promoter region. NFAT activation increased in AF fibroblasts, and NFAT negatively regulated microRNA-26 transcription. In vivo pyrazole-3 administration suppressed AF while decreasing fibroblast proliferation and extracellular matrix gene expression. CONCLUSIONS-: TRPC3 channels regulate cardiac fibroblast proliferation and differentiation, likely by controlling the Ca influx that activates extracellular signal-regulated kinase signaling. AF increases TRPC3 channel expression by causing NFAT-mediated downregulation of microRNA-26 and causes TRPC3-dependent enhancement of fibroblast proliferation and differentiation. In vivo, TRPC3 blockade prevents AF substrate development in a dog model of electrically maintained AF. TRPC3 likely plays an important role in AF by promoting fibroblast pathophysiology and is a novel potential therapeutic target.

Original languageEnglish
Pages (from-to)2051-2064
Number of pages14
JournalCirculation
Volume126
Issue number17
DOIs
Publication statusPublished - 23-10-2012

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Atrial Fibrillation
Fibroblasts
Extracellular Signal-Regulated MAP Kinases
NFATC Transcription Factors
MicroRNAs
Phosphorylation
Dogs
Extracellular Matrix
Smooth Muscle
Canidae
Actins
Down-Regulation
Gene Expression
Muscle Proteins
Genetic Promoter Regions
Goats
Angiotensin II
Small Interfering RNA
Cations
Up-Regulation

All Science Journal Classification (ASJC) codes

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Harada, Masahide ; Luo, Xiaobin ; Qi, Xiao Yan ; Tadevosyan, Artavazd ; Maguy, Ange ; Ordog, Balazs ; Ledoux, Jonathan ; Kato, Takeshi ; Naud, Patrice ; Voigt, Niels ; Shi, Yanfen ; Kamiya, Kaichiro ; Murohara, Toyoaki ; Kodama, Itsuo ; Tardif, Jean Claude ; Schotten, Ulrich ; Van Wagoner, David R. ; Dobrev, Dobromir ; Nattel, Stanley. / Transient receptor potential canonical-3 channel-dependent fibroblast regulation in atrial fibrillation. In: Circulation. 2012 ; Vol. 126, No. 17. pp. 2051-2064.
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abstract = "BACKGROUND-: Fibroblast proliferation and differentiation are central in atrial fibrillation (AF)-promoting remodeling. Here, we investigated fibroblast regulation by Ca-permeable transient receptor potential canonical-3 (TRPC3) channels. METHODS AND RESULTS-: Freshly isolated rat cardiac fibroblasts abundantly expressed TRPC3 and had appreciable nonselective cation currents (INSC) sensitive to a selective TPRC3 channel blocker, pyrazole-3 (3 μmol/L). Pyrazole-3 suppressed angiotensin II-induced Ca influx, proliferation, and α-smooth muscle actin protein expression in fibroblasts. Ca removal and TRPC3 blockade suppressed extracellular signal-regulated kinase phosphorylation, and extracellular signal-regulated kinase phosphorylation inhibition reduced fibroblast proliferation. TRPC3 expression was upregulated in atria from AF patients, goats with electrically maintained AF, and dogs with tachypacing-induced heart failure. TRPC3 knockdown (based on short hairpin RNA [shRNA]) decreased canine atrial fibroblast proliferation. In left atrial fibroblasts freshly isolated from dogs kept in AF for 1 week by atrial tachypacing, TRPC3 protein expression, currents, extracellular signal-regulated kinase phosphorylation, and extracellular matrix gene expression were all significantly increased. In cultured left atrial fibroblasts from AF dogs, proliferation rates, α-smooth muscle actin expression, and extracellular signal-regulated kinase phosphorylation were increased and were suppressed by pyrazole-3. MicroRNA-26 was downregulated in canine AF atria; experimental microRNA-26 knockdown reproduced AF-induced TRPC3 upregulation and fibroblast activation. MicroRNA-26 has NFAT (nuclear factor of activated T cells) binding sites in the 5′ promoter region. NFAT activation increased in AF fibroblasts, and NFAT negatively regulated microRNA-26 transcription. In vivo pyrazole-3 administration suppressed AF while decreasing fibroblast proliferation and extracellular matrix gene expression. CONCLUSIONS-: TRPC3 channels regulate cardiac fibroblast proliferation and differentiation, likely by controlling the Ca influx that activates extracellular signal-regulated kinase signaling. AF increases TRPC3 channel expression by causing NFAT-mediated downregulation of microRNA-26 and causes TRPC3-dependent enhancement of fibroblast proliferation and differentiation. In vivo, TRPC3 blockade prevents AF substrate development in a dog model of electrically maintained AF. TRPC3 likely plays an important role in AF by promoting fibroblast pathophysiology and is a novel potential therapeutic target.",
author = "Masahide Harada and Xiaobin Luo and Qi, {Xiao Yan} and Artavazd Tadevosyan and Ange Maguy and Balazs Ordog and Jonathan Ledoux and Takeshi Kato and Patrice Naud and Niels Voigt and Yanfen Shi and Kaichiro Kamiya and Toyoaki Murohara and Itsuo Kodama and Tardif, {Jean Claude} and Ulrich Schotten and {Van Wagoner}, {David R.} and Dobromir Dobrev and Stanley Nattel",
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Harada, M, Luo, X, Qi, XY, Tadevosyan, A, Maguy, A, Ordog, B, Ledoux, J, Kato, T, Naud, P, Voigt, N, Shi, Y, Kamiya, K, Murohara, T, Kodama, I, Tardif, JC, Schotten, U, Van Wagoner, DR, Dobrev, D & Nattel, S 2012, 'Transient receptor potential canonical-3 channel-dependent fibroblast regulation in atrial fibrillation', Circulation, vol. 126, no. 17, pp. 2051-2064. https://doi.org/10.1161/CIRCULATIONAHA.112.121830

Transient receptor potential canonical-3 channel-dependent fibroblast regulation in atrial fibrillation. / Harada, Masahide; Luo, Xiaobin; Qi, Xiao Yan; Tadevosyan, Artavazd; Maguy, Ange; Ordog, Balazs; Ledoux, Jonathan; Kato, Takeshi; Naud, Patrice; Voigt, Niels; Shi, Yanfen; Kamiya, Kaichiro; Murohara, Toyoaki; Kodama, Itsuo; Tardif, Jean Claude; Schotten, Ulrich; Van Wagoner, David R.; Dobrev, Dobromir; Nattel, Stanley.

In: Circulation, Vol. 126, No. 17, 23.10.2012, p. 2051-2064.

Research output: Contribution to journalArticle

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T1 - Transient receptor potential canonical-3 channel-dependent fibroblast regulation in atrial fibrillation

AU - Harada, Masahide

AU - Luo, Xiaobin

AU - Qi, Xiao Yan

AU - Tadevosyan, Artavazd

AU - Maguy, Ange

AU - Ordog, Balazs

AU - Ledoux, Jonathan

AU - Kato, Takeshi

AU - Naud, Patrice

AU - Voigt, Niels

AU - Shi, Yanfen

AU - Kamiya, Kaichiro

AU - Murohara, Toyoaki

AU - Kodama, Itsuo

AU - Tardif, Jean Claude

AU - Schotten, Ulrich

AU - Van Wagoner, David R.

AU - Dobrev, Dobromir

AU - Nattel, Stanley

PY - 2012/10/23

Y1 - 2012/10/23

N2 - BACKGROUND-: Fibroblast proliferation and differentiation are central in atrial fibrillation (AF)-promoting remodeling. Here, we investigated fibroblast regulation by Ca-permeable transient receptor potential canonical-3 (TRPC3) channels. METHODS AND RESULTS-: Freshly isolated rat cardiac fibroblasts abundantly expressed TRPC3 and had appreciable nonselective cation currents (INSC) sensitive to a selective TPRC3 channel blocker, pyrazole-3 (3 μmol/L). Pyrazole-3 suppressed angiotensin II-induced Ca influx, proliferation, and α-smooth muscle actin protein expression in fibroblasts. Ca removal and TRPC3 blockade suppressed extracellular signal-regulated kinase phosphorylation, and extracellular signal-regulated kinase phosphorylation inhibition reduced fibroblast proliferation. TRPC3 expression was upregulated in atria from AF patients, goats with electrically maintained AF, and dogs with tachypacing-induced heart failure. TRPC3 knockdown (based on short hairpin RNA [shRNA]) decreased canine atrial fibroblast proliferation. In left atrial fibroblasts freshly isolated from dogs kept in AF for 1 week by atrial tachypacing, TRPC3 protein expression, currents, extracellular signal-regulated kinase phosphorylation, and extracellular matrix gene expression were all significantly increased. In cultured left atrial fibroblasts from AF dogs, proliferation rates, α-smooth muscle actin expression, and extracellular signal-regulated kinase phosphorylation were increased and were suppressed by pyrazole-3. MicroRNA-26 was downregulated in canine AF atria; experimental microRNA-26 knockdown reproduced AF-induced TRPC3 upregulation and fibroblast activation. MicroRNA-26 has NFAT (nuclear factor of activated T cells) binding sites in the 5′ promoter region. NFAT activation increased in AF fibroblasts, and NFAT negatively regulated microRNA-26 transcription. In vivo pyrazole-3 administration suppressed AF while decreasing fibroblast proliferation and extracellular matrix gene expression. CONCLUSIONS-: TRPC3 channels regulate cardiac fibroblast proliferation and differentiation, likely by controlling the Ca influx that activates extracellular signal-regulated kinase signaling. AF increases TRPC3 channel expression by causing NFAT-mediated downregulation of microRNA-26 and causes TRPC3-dependent enhancement of fibroblast proliferation and differentiation. In vivo, TRPC3 blockade prevents AF substrate development in a dog model of electrically maintained AF. TRPC3 likely plays an important role in AF by promoting fibroblast pathophysiology and is a novel potential therapeutic target.

AB - BACKGROUND-: Fibroblast proliferation and differentiation are central in atrial fibrillation (AF)-promoting remodeling. Here, we investigated fibroblast regulation by Ca-permeable transient receptor potential canonical-3 (TRPC3) channels. METHODS AND RESULTS-: Freshly isolated rat cardiac fibroblasts abundantly expressed TRPC3 and had appreciable nonselective cation currents (INSC) sensitive to a selective TPRC3 channel blocker, pyrazole-3 (3 μmol/L). Pyrazole-3 suppressed angiotensin II-induced Ca influx, proliferation, and α-smooth muscle actin protein expression in fibroblasts. Ca removal and TRPC3 blockade suppressed extracellular signal-regulated kinase phosphorylation, and extracellular signal-regulated kinase phosphorylation inhibition reduced fibroblast proliferation. TRPC3 expression was upregulated in atria from AF patients, goats with electrically maintained AF, and dogs with tachypacing-induced heart failure. TRPC3 knockdown (based on short hairpin RNA [shRNA]) decreased canine atrial fibroblast proliferation. In left atrial fibroblasts freshly isolated from dogs kept in AF for 1 week by atrial tachypacing, TRPC3 protein expression, currents, extracellular signal-regulated kinase phosphorylation, and extracellular matrix gene expression were all significantly increased. In cultured left atrial fibroblasts from AF dogs, proliferation rates, α-smooth muscle actin expression, and extracellular signal-regulated kinase phosphorylation were increased and were suppressed by pyrazole-3. MicroRNA-26 was downregulated in canine AF atria; experimental microRNA-26 knockdown reproduced AF-induced TRPC3 upregulation and fibroblast activation. MicroRNA-26 has NFAT (nuclear factor of activated T cells) binding sites in the 5′ promoter region. NFAT activation increased in AF fibroblasts, and NFAT negatively regulated microRNA-26 transcription. In vivo pyrazole-3 administration suppressed AF while decreasing fibroblast proliferation and extracellular matrix gene expression. CONCLUSIONS-: TRPC3 channels regulate cardiac fibroblast proliferation and differentiation, likely by controlling the Ca influx that activates extracellular signal-regulated kinase signaling. AF increases TRPC3 channel expression by causing NFAT-mediated downregulation of microRNA-26 and causes TRPC3-dependent enhancement of fibroblast proliferation and differentiation. In vivo, TRPC3 blockade prevents AF substrate development in a dog model of electrically maintained AF. TRPC3 likely plays an important role in AF by promoting fibroblast pathophysiology and is a novel potential therapeutic target.

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