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Mélanie Lambert - One of the best experts on this subject based on the ideXlab platform.
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KCNK3 Dysfunction Exaggerates The Development Of Pulmonary Hypertension Induced By Left Ventricular Pressure Overload.
Cardiovascular research, 2021Co-Authors: Mélanie Lambert, Catherine Rucker-martin, P. Mendes-ferreira, Maria-rosa Ghigna, Hélène Leribeuz, Rui Adão, Angèle Boet, Véronique Capuano, Carmen Brás-silva, Rozenn QuarckAbstract:Aims Pulmonary hypertension (PH) is a common complication of left heart disease (LHD, group 2 PH) leading to right ventricular (RV) failure and death. Several loss-of-function (LOF) mutations in KCNK3 were identified in pulmonary arterial hypertension (PAH, group 1 PH). Additionally, we found that KCNK3 dysfunction is a hallmark of PAH at pulmonary vascular and RV levels. However, the role of KCNK3 in the pathobiology of PH due to LHD is unknown. Methods and results We evaluated the role of KCNK3 on PH induced by ascending aortic constriction (AAC), in WT and KCNK3-LOF-mutated rats, by echocardiography, RV catheterization, histology analyses, and molecular biology experiments. We found that KCNK3-LOF-mutation had no consequence on the development of left ventricular (LV) compensated concentric hypertrophy in AAC, while left atrial (LA) emptying fraction was impaired in AAC-KCNK3-mutated rats. AAC-animals (WT and KCNK3-mutated rats) developed PH secondary to AAC and KCNK3-mutated rats developed more severe PH than WT. AAC-KCNK3-mutated rats developed RV and LV fibrosis in association with an increase of Col1a1 mRNA in RV and LV. AAC-KCNK3-mutated rats developed severe pulmonary vascular (pulmonary artery as well as pulmonary veins) remodelling with intense peri-vascular and peri-bronchial inflammation, perivascular edema, alveolar wall thickening, and exaggerated lung vascular cell proliferation compared to AAC-WT-rats. Finally, in lung, RV, LV, and LA of AAC-KCNK3-mutated rats, we found a strong increased expression of Il-6 and periostin expression and a reduction of lung Ctnnd1 mRNA (coding for p120 catenin), contributing to the exaggerated pulmonary and heart remodelling and pulmonary vascular edema in AAC-KCNK3-mutated rats. Conclusions Our results indicate that KCNK3-LOF is a key event in the pathobiology of PH due to AAC, suggesting that KCNK3 channel dysfunction could play a potential key role in the development of PH due to LHD.
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Proteomic Analysis of KCNK3 Loss of Expression Identified Dysregulated Pathways in Pulmonary Vascular Cells.
International journal of molecular sciences, 2020Co-Authors: Hélène Le Ribeuz, Mélanie Lambert, Barbara Girerd, Florent Dumont, Guillaume Ruellou, Thierry Balliau, Marceau Quatredeniers, Sylvia Cohen-kaminsky, Olaf Mercier, Stéphanie Yen-nicolaÿAbstract:The physiopathology of pulmonary arterial hypertension (PAH) is characterized by pulmonary artery smooth muscle cell (PASMC) and endothelial cell (PAEC) dysfunction, contributing to pulmonary arterial obstruction and PAH progression. KCNK3 loss of function mutations are responsible for the first channelopathy identified in PAH. Loss of KCNK3 function/expression is a hallmark of PAH. However, the molecular mechanisms involved in KCNK3 dysfunction are mostly unknown. To identify the pathological molecular mechanisms downstream of KCNK3 in human PASMCs (hPASMCs) and human PAECs (hPAECs), we used a Liquid Chromatography-Tandem Mass Spectrometry-based proteomic approach to identify the molecular pathways regulated by KCNK3. KCNK3 loss of expression was induced in control hPASMCs or hPAECs by specific siRNA targeting KCNK3. We found that the loss of KCNK3 expression in hPAECs and hPASMCs leads to 326 and 222 proteins differentially expressed, respectively. Among them, 53 proteins were common to hPAECs and hPASMCs. The specific proteome remodeling in hPAECs in absence of KCNK3 was mostly related to the activation of glycolysis, the superpathway of methionine degradation, and the mTOR signaling pathways, and to a reduction in EIF2 signaling pathways. In hPASMCs, we found an activation of the PI3K/AKT signaling pathways and a reduction in EIF2 signaling and the Purine Nucleotides De Novo Biosynthesis II and IL-8 signaling pathways. Common to hPAECs and hPASMCs, we found that the loss of KCNK3 expression leads to the activation of the NRF2-mediated oxidative stress response and a reduction in the interferon pathway. In the hPAECs and hPASMCs, we found an increased expression of HO-1 (heme oxygenase-1) and a decreased IFIT3 (interferon-induced proteins with tetratricopeptide repeats 3) (confirmed by Western blotting), allowing us to identify these axes to understand the consequences of KCNK3 dysfunction. Our experiments, based on the loss of KCNK3 expression by a specific siRNA strategy in control hPAECs and hPASMCs, allow us to identify differences in the activation of several signaling pathways, indicating the key role played by KCNK3 dysfunction in the development of PAH. Altogether, these results allow us to better understand the consequences of KCNK3 dysfunction and suggest that KCNK3 loss of expression acts in favor of the proliferation and migration of hPASMCs and promotes the metabolic shift and apoptosis resistance of hPAECs.
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In vivo miR-138-5p inhibition alleviates monocrotaline-induced pulmonary hypertension and normalizes pulmonary KCNK3 and SLC45A3 expression
Respiratory research, 2020Co-Authors: Hélène Le Ribeuz, Mélanie Lambert, Aurélie Hautefort, David Montani, Marc Humbert, Maria-rosa Ghigna, Sylvia Cohen-kaminsky, Frédéric Perros, Audrey Courboulin, Fabrice AntignyAbstract:Background The pathogenesis of pulmonary arterial hypertension (PAH) involves many signalling pathways. MicroRNAs are potential candidates involved in simultaneously coordinating multiple genes under such multifactorial conditions. Methods and results MiR-138-5p is overexpressed in pulmonary arterial smooth muscle cells (PASMCs) from PAH patients and in lungs from rats with monocrotaline-induced pulmonary hypertension (MCT-PH). MiR-138-5p is predicted to regulate the expression of the potassium channel KCNK3, whose loss is associated with the development and progression of PAH. We hypothesized that, in vivo, miR-138-5p inhibition would restore KCNK3 lung expression and subsequently alleviate PAH. Nebulization-based delivery of anti-miR-138-5p to rats with established MCT-PH significantly reduced the right ventricular systolic pressure and significantly improved the pulmonary arterial acceleration time (PAAT). These haemodynamic improvements were related to decrease pulmonary vascular remodelling, lung inflammation and pulmonary vascular cell proliferation in situ. In vivo inhibition of miR-138-5p restored KCNK3 mRNA expression and SLC45A3 protein expression in the lungs. Conclusions We confirmed that in vivo inhibition of miR-138-5p reduces the development of PH in experimental MCT-PH. The possible curative mechanisms involve at least the normalization of lung KCNK3 as well as SLC45A3 expression.
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etude de l implication de la dysfonction de KCNK3 dans le developpement de l hypertension arterielle pulmonaire
2019Co-Authors: Mélanie LambertAbstract:L’hypertension arterielle pulmonaire (HTAP) est une maladie rare resultant de l’obstruction progressive des petites arteres pulmonaires, via un remodelage de la paroi vasculaire, associee a une vasoconstriction entrainant une hypertrophie puis une defaillance cardiaque droite qui aboutit au deces du patient. Depuis 2013, 10 mutations, dans le gene KCNK3 (codant pour un canal potassique), ont ete identifiees chez des patients atteints d’HTAP. Toutes les mutations identifiees entrainent une perte de fonction du canal. De plus, notre equipe a pu demontrer que la perte de KCNK3 est commune a toutes les formes d’HTAP : idiopathique et heritable ainsi qu’experimentale. Durant ma these, mon projet principal a donc ete d’etudier l’implication de la dysfonction de KCNK3 dans le developpement de l’HTAP. Tout d’abord, nous avons pu demontrer que la perte d’expression/fonction de KCNK3 est egalement une caracteristique de l’hypertrophie / la dysfonction ventriculaire droite. Par la suite, en caracterisant un modele unique de rats mutes pour le gene KCNK3, nous avons pu demontrer que l'inactivation genetique de KCNK3 chez le rat conduit a une alteration vasculaire pulmonaire facilitant ainsi le developpement d’une hypertension pulmonaire (HTP). Faisant de ce modele un nouvel outil permettant de comprendre les mecanismes initiateurs de l’HTP et representerai un outil pertinent pour developper des cibles therapeutiques. Pour finir, nous avons montre que le developpement d’une HTP due a une insuffisance cardiaque gauche (via une ligature du l’aorte) est facilitee chez nos rats mutes pour KCNK3.
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Characterization of KCNK3 -Mutated Rat, a Novel Model of Pulmonary Hypertension
Circulation Research, 2019Co-Authors: Mélanie Lambert, Christine Péchoux, Angèle Boet, Véronique Capuano, Laurent Tesson, Thomas Bertero, Morad Nakhleh, Séverine Remy, Ignacio Anegon, Aurélie HautefortAbstract:RATIONALE: Pulmonary arterial hypertension is a severe lethal cardiopulmonary disease. Loss of function mutations in KCNK3 (potassium channel subfamily K member 3) gene, which encodes an outward rectifier K + channel, have been identified in pulmonary arterial hypertension patients. OBJECTIVE: We have demonstrated that KCNK3 dysfunction is common to heritable and nonheritable pulmonary arterial hypertension and to experimental pulmonary hypertension (PH). Finally, KCNK3 is not functional in mouse pulmonary vasculature. METHODS AND RESULTS: Using CRISPR/Cas9 technology, we generated a 94 bp out of frame deletion in exon 1 of KCNK3 gene and characterized these rats at the electrophysiological, echocardiographic, hemodynamic, morphological, cellular, and molecular levels to decipher the cellular mechanisms associated with loss of KCNK3. Using patch-clamp technique, we validated our transgenic strategy by demonstrating the absence of KCNK3 current in freshly isolated pulmonary arterial smooth muscle cells from KCNK3-mutated rats. At 4 months of age, echocardiographic parameters revealed shortening of the pulmonary artery acceleration time associated with elevation of the right ventricular systolic pressure. KCNK3-mutated rats developed more severe PH than wild-type rats after monocrotaline exposure or chronic hypoxia exposure. KCNK3-mutation induced a lung distal neomuscularization and perivascular extracellular matrix activation. Lungs of KCNK3-mutated rats were characterized by overactivation of ERK1/2 (extracellular signal-regulated kinase1-/2), AKT (protein kinase B), SRC, and overexpression of HIF1-α (hypoxia-inducible factor-1 α), survivin, and VWF (Von Willebrand factor). Linked with plasma membrane depolarization, reduced endothelial-NOS expression and desensitization of endothelial-derived hyperpolarizing factor, KCNK3-mutated rats presented predisposition to vasoconstriction of pulmonary arteries and a severe loss of sildenafil-induced pulmonary arteries relaxation. Moreover, we showed strong alteration of right ventricular cardiomyocyte excitability. Finally, KCNK3-mutated rats developed age-dependent PH associated with low serum-albumin concentration. CONCLUSIONS: We established the first KCNK3-mutated rat model of PH. Our results confirm that KCNK3 loss of function is a key event in pulmonary arterial hypertension pathogenesis. This model presents new opportunities for understanding the initiating mechanisms of PH and testing biologically relevant therapeutic molecules in the context of PH. VISUAL OVERVIEW: An online visual overview is available for this article. P ulmonary arterial hypertension (PAH) is an uncommon , progressive, and severe disease with an estimated prevalence of 15 to 50 per million. 1 PAH has been hemodynamically defined by an elevation of the mean pulmonary artery pressure >20 mm Hg and pulmonary vascular resistance >3 Wood units at rest. 2 PAH results from increased pulmonary vascular resistance because of remodeling of the small distal pulmonary arteries (PAs) and arterioles (diameter
Fabrice Antigny - One of the best experts on this subject based on the ideXlab platform.
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Implication of Potassium Channels in the Pathophysiology of Pulmonary Arterial Hypertension
Biomolecules, 2020Co-Authors: Hélène Le Ribeuz, Barbara Girerd, David Montani, Marc Humbert, Véronique Capuano, Fabrice AntignyAbstract:Pulmonary arterial hypertension (PAH) is a rare and severe cardiopulmonary disease without curative treatments. PAH is a multifactorial disease that involves genetic predisposition, epigenetic factors, and environmental factors (drugs, toxins, viruses, hypoxia, and inflammation), which contribute to the initiation or development of irreversible remodeling of the pulmonary vessels. The recent identification of loss-of-function mutations in KCNK3 (KCNK3 or TASK-1) and ABCC8 (SUR1), or gain-of-function mutations in ABCC9 (SUR2), as well as polymorphisms in KCNA5 (Kv1.5), which encode two potassium (K+) channels and two K+ channel regulatory subunits, has revived the interest of ion channels in PAH. This review focuses on KCNK3, SUR1, SUR2, and Kv1.5 channels in pulmonary vasculature and discusses their pathophysiological contribution to and therapeutic potential in PAH.
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In vivo miR-138-5p inhibition alleviates monocrotaline-induced pulmonary hypertension and normalizes pulmonary KCNK3 and SLC45A3 expression
Respiratory research, 2020Co-Authors: Hélène Le Ribeuz, Mélanie Lambert, Aurélie Hautefort, David Montani, Marc Humbert, Maria-rosa Ghigna, Sylvia Cohen-kaminsky, Frédéric Perros, Audrey Courboulin, Fabrice AntignyAbstract:Background The pathogenesis of pulmonary arterial hypertension (PAH) involves many signalling pathways. MicroRNAs are potential candidates involved in simultaneously coordinating multiple genes under such multifactorial conditions. Methods and results MiR-138-5p is overexpressed in pulmonary arterial smooth muscle cells (PASMCs) from PAH patients and in lungs from rats with monocrotaline-induced pulmonary hypertension (MCT-PH). MiR-138-5p is predicted to regulate the expression of the potassium channel KCNK3, whose loss is associated with the development and progression of PAH. We hypothesized that, in vivo, miR-138-5p inhibition would restore KCNK3 lung expression and subsequently alleviate PAH. Nebulization-based delivery of anti-miR-138-5p to rats with established MCT-PH significantly reduced the right ventricular systolic pressure and significantly improved the pulmonary arterial acceleration time (PAAT). These haemodynamic improvements were related to decrease pulmonary vascular remodelling, lung inflammation and pulmonary vascular cell proliferation in situ. In vivo inhibition of miR-138-5p restored KCNK3 mRNA expression and SLC45A3 protein expression in the lungs. Conclusions We confirmed that in vivo inhibition of miR-138-5p reduces the development of PH in experimental MCT-PH. The possible curative mechanisms involve at least the normalization of lung KCNK3 as well as SLC45A3 expression.
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Abstract 15397: KCNK3 Channel Inactivation Leads to Pulmonary Vascular Alterations in Rat
Circulation, 2017Co-Authors: Mélanie Lambert, Aurélie Hautefort, Boris Manoury, David Montani, Marc Humbert, Frédéric Perros, Fabrice AntignyAbstract:Background: Pulmonary arterial hypertension (PAH) is a severe and lethal cardio-pulmonary disease characterized by a progressive occlusion of the distal pulmonary arteries. Mutations in the KCNK3 g...
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Abstract 17279: KCNK3 Dysfunction Contributes to the Development of Pulmonary Arterial Hypertension - Characterization of KCNK3 Deficient Rats
Circulation, 2016Co-Authors: Fabrice Antigny, Mélanie Lambert, Aurélie Hautefort, Boris Manoury, Philippe Jourdon, Catherine Rucker-martin, Barbara Girerd, David Montani, Gérald Simonnneau, Marc HumbertAbstract:Background: Inactivating mutations in the KCNK3 gene (K + channel) have been identified in heritable forms of pulmonary arterial hypertension (PAH). We discovered that KCNK3 dysfunction contributes actually to the development of both heritable and non-heritable PAH, and to experimental pulmonary hypertension (PH) ( Antigny et al. Circulation 2016 ). Methods and results: We generated KCNK3 deficient rats using CRISPR-Cas9 technology and characterized at electrophysiological, hemodynamics, morphological and molecular levels the first genetically modified rat model linked to KCNK3 mutation: KCNK3 Δ94Ex1/+ (94pb deletion in exon 1 of KCNK3 gene). Using a patch-clamp technique in freshly isolated pulmonary artery smooth muscle cells (PASMCs), we found that KCNK3-current is absent in heterozygous and homozygous KCNK3 mutated rats and that isolated PASMC are significantly depolarized compared to WT rats. KCNK3 mutation induced distal neomuscularization, abnormal pulmonary arteries vasoreactivity, elevated mean right ventricular systolic pressures and upregulation of MAP kinase signaling pathways. Heterozygous KCNK3 -mutated rats developed more severe pulmonary hypertension than WT littermate under hypoxic condition. Conclusions: In this study we established the first KCNK3 deficient rat model. We demonstrated that a genetic inactivation of KCNK3 in rats promoted the early signs of pulmonary hypertension, confirming that KCNK3 loss of function is a key event in PAH pathogenesis. The development and characterization of the first KCNK3 deficient PH-rats model open new opportunities for testing relevant therapeutics molecules in context of heritable PH.
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LATE-BREAKING ABSTRACT: KCNK3 dysfunction contributes to the development of pulmonary arterial hypertension – Characterization of KCNK3 deficient rats
4.3 Pulmonary Circulation and Pulmonary Vascular Diseases, 2016Co-Authors: Fabrice Antigny, Mélanie Lambert, Aurélie Hautefort, Boris Manoury, Philippe Jourdon, Catherine Rucker-martin, Barbara Girerd, David Montani, Gérald Simonneau, Mars HumbertAbstract:Background: Inactivating mutations in the KCNK3 gene (K + channel) have been identified in heritable forms of pulmonary arterial hypertension (PAH). We discovered that KCNK3 dysfunction contributes actually to the development of both heritable and non-heritable PAH, and to experimental pulmonary hypertension (PH) ( Antigny et al. Circulation 2016 ). Methods and results: We generated KCNK3 deficient rats using CRISPR-Cas9 technology and characterized at electrophysiological, hemodynamics, morphological and molecular levels the first genetically modified rat model linked to KCNK3 mutation: KCNK3 Δ 94Ex1/+ (94pb deletion in exon 1 of KCNK3 gene). Using a patch–clamp technique in freshly isolated pulmonary artery smooth muscle cells (PASMCs), we found that KCNK3-current is absent in heterozygous and homozygous KCNK3 mutated rats and that isolated PASMC are significantly depolarized compared to WT rats. KCNK3 mutation induced distal neomuscularization, abnormal pulmonary arteries vasoreactivity, elevated mean right ventricular systolic pressures and upregulation of MAP kinase signaling pathways. Heterozygous KCNK3 -mutated rats developed more severe pulmonary hypertension than WT littermate under hypoxic condition. Conclusions: In this study we established the first KCNK3 deficient rat model. We demonstrated that a genetic inactivation of KCNK3 in rats promoted the early signs of pulmonary hypertension, confirming that KCNK3 loss of function is a key event in PAH pathogenesis. The development and characterization of the first KCNK3 deficient PH-rats model open new opportunities for testing relevant therapeutics molecules in context of heritable PH.
Aurélie Hautefort - One of the best experts on this subject based on the ideXlab platform.
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In vivo miR-138-5p inhibition alleviates monocrotaline-induced pulmonary hypertension and normalizes pulmonary KCNK3 and SLC45A3 expression
Respiratory research, 2020Co-Authors: Hélène Le Ribeuz, Mélanie Lambert, Aurélie Hautefort, David Montani, Marc Humbert, Maria-rosa Ghigna, Sylvia Cohen-kaminsky, Frédéric Perros, Audrey Courboulin, Fabrice AntignyAbstract:Background The pathogenesis of pulmonary arterial hypertension (PAH) involves many signalling pathways. MicroRNAs are potential candidates involved in simultaneously coordinating multiple genes under such multifactorial conditions. Methods and results MiR-138-5p is overexpressed in pulmonary arterial smooth muscle cells (PASMCs) from PAH patients and in lungs from rats with monocrotaline-induced pulmonary hypertension (MCT-PH). MiR-138-5p is predicted to regulate the expression of the potassium channel KCNK3, whose loss is associated with the development and progression of PAH. We hypothesized that, in vivo, miR-138-5p inhibition would restore KCNK3 lung expression and subsequently alleviate PAH. Nebulization-based delivery of anti-miR-138-5p to rats with established MCT-PH significantly reduced the right ventricular systolic pressure and significantly improved the pulmonary arterial acceleration time (PAAT). These haemodynamic improvements were related to decrease pulmonary vascular remodelling, lung inflammation and pulmonary vascular cell proliferation in situ. In vivo inhibition of miR-138-5p restored KCNK3 mRNA expression and SLC45A3 protein expression in the lungs. Conclusions We confirmed that in vivo inhibition of miR-138-5p reduces the development of PH in experimental MCT-PH. The possible curative mechanisms involve at least the normalization of lung KCNK3 as well as SLC45A3 expression.
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Characterization of KCNK3 -Mutated Rat, a Novel Model of Pulmonary Hypertension
Circulation Research, 2019Co-Authors: Mélanie Lambert, Christine Péchoux, Angèle Boet, Véronique Capuano, Laurent Tesson, Thomas Bertero, Morad Nakhleh, Séverine Remy, Ignacio Anegon, Aurélie HautefortAbstract:RATIONALE: Pulmonary arterial hypertension is a severe lethal cardiopulmonary disease. Loss of function mutations in KCNK3 (potassium channel subfamily K member 3) gene, which encodes an outward rectifier K + channel, have been identified in pulmonary arterial hypertension patients. OBJECTIVE: We have demonstrated that KCNK3 dysfunction is common to heritable and nonheritable pulmonary arterial hypertension and to experimental pulmonary hypertension (PH). Finally, KCNK3 is not functional in mouse pulmonary vasculature. METHODS AND RESULTS: Using CRISPR/Cas9 technology, we generated a 94 bp out of frame deletion in exon 1 of KCNK3 gene and characterized these rats at the electrophysiological, echocardiographic, hemodynamic, morphological, cellular, and molecular levels to decipher the cellular mechanisms associated with loss of KCNK3. Using patch-clamp technique, we validated our transgenic strategy by demonstrating the absence of KCNK3 current in freshly isolated pulmonary arterial smooth muscle cells from KCNK3-mutated rats. At 4 months of age, echocardiographic parameters revealed shortening of the pulmonary artery acceleration time associated with elevation of the right ventricular systolic pressure. KCNK3-mutated rats developed more severe PH than wild-type rats after monocrotaline exposure or chronic hypoxia exposure. KCNK3-mutation induced a lung distal neomuscularization and perivascular extracellular matrix activation. Lungs of KCNK3-mutated rats were characterized by overactivation of ERK1/2 (extracellular signal-regulated kinase1-/2), AKT (protein kinase B), SRC, and overexpression of HIF1-α (hypoxia-inducible factor-1 α), survivin, and VWF (Von Willebrand factor). Linked with plasma membrane depolarization, reduced endothelial-NOS expression and desensitization of endothelial-derived hyperpolarizing factor, KCNK3-mutated rats presented predisposition to vasoconstriction of pulmonary arteries and a severe loss of sildenafil-induced pulmonary arteries relaxation. Moreover, we showed strong alteration of right ventricular cardiomyocyte excitability. Finally, KCNK3-mutated rats developed age-dependent PH associated with low serum-albumin concentration. CONCLUSIONS: We established the first KCNK3-mutated rat model of PH. Our results confirm that KCNK3 loss of function is a key event in pulmonary arterial hypertension pathogenesis. This model presents new opportunities for understanding the initiating mechanisms of PH and testing biologically relevant therapeutic molecules in the context of PH. VISUAL OVERVIEW: An online visual overview is available for this article. P ulmonary arterial hypertension (PAH) is an uncommon , progressive, and severe disease with an estimated prevalence of 15 to 50 per million. 1 PAH has been hemodynamically defined by an elevation of the mean pulmonary artery pressure >20 mm Hg and pulmonary vascular resistance >3 Wood units at rest. 2 PAH results from increased pulmonary vascular resistance because of remodeling of the small distal pulmonary arteries (PAs) and arterioles (diameter
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Characterization of KCNK3-Mutated Rat, a Novel Model of Pulmonary Hypertension.
Circulation research, 2019Co-Authors: Mélanie Lambert, Christine Péchoux, Angèle Boet, Véronique Capuano, Laurent Tesson, Thomas Bertero, Morad Nakhleh, Séverine Remy, Ignacio Anegon, Aurélie HautefortAbstract:Rationale: Pulmonary arterial hypertension is a severe lethal cardiopulmonary disease. Loss of function mutations in KCNK3 (potassium channel subfamily K member 3) gene, which encodes an outward re...
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Loss of KCNK3 is a hallmark of RV hypertrophy/dysfunction associated with pulmonary hypertension.
Cardiovascular research, 2018Co-Authors: Mélanie Lambert, Aurélie Hautefort, Catherine Rucker-martin, P. Mendes-ferreira, Rui Adão, Angèle Boet, Véronique Capuano, Carmen Brás-silva, Stéphane N. Hatem, Jean-baptiste MichelAbstract:Aims Mutations in the KCNK3 gene, which encodes for an outward-rectifier K+ channel, have been identified in patients suffering from pulmonary arterial hypertension (PAH), and constitute the first described channelopathy in PAH. In human PAH and experimental pulmonary hypertension (PH), we demonstrated that KCNK3 expression and function are severely reduced in pulmonary vascular cells, promoting PH-like phenotype at the morphologic and haemodynamic levels. Since KCNK3 channel is also expressed in both the human and rodent heart, we aimed to elucidate the pathophysiological role of KCNK3 channel in right ventricular (RV) hypertrophy (RVH) related to PH. Methods and results Using whole-cell Patch-clamp technique, we demonstrated that KCNK3 is predominantly expressed in adult rat RV cardiomyocytes compared to the left ventricle cardiomyocytes and participates in the repolarizing phase of the RV action potential. We revealed a reduction in KCNK3 function prior to development of RVH and the rise of pulmonary vascular resistance. KCNK3 function is severely reduced in RV cardiomyocytes during the development of RVH in several rat models of PH (exposure to monocrotaline, chronic hypoxia, and Sugen/hypoxia) and chronic RV pressure overload (pulmonary artery banding). In experimental PH, we revealed a reduction in KCNK3 function before any rise in pulmonary vascular resistance and the development of RVH. KCNK3 mRNA level is also reduced in human RV tissues from PAH patients compared to non-PAH patients. In line with these findings, chronic inhibition of KCNK3 in rats with the specific inhibitor (A293) induces RV hypertrophy which is associated with the re-expression of foetal genes, RV fibrosis, RV inflammation, and subsequent loss of RV performance as assessed by echocardiography. Conclusion Our data indicate that loss of KCNK3 function and expression is a hallmark of the RV hypertrophy/dysfunction associated with PH.
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Abstract 15397: KCNK3 Channel Inactivation Leads to Pulmonary Vascular Alterations in Rat
Circulation, 2017Co-Authors: Mélanie Lambert, Aurélie Hautefort, Boris Manoury, David Montani, Marc Humbert, Frédéric Perros, Fabrice AntignyAbstract:Background: Pulmonary arterial hypertension (PAH) is a severe and lethal cardio-pulmonary disease characterized by a progressive occlusion of the distal pulmonary arteries. Mutations in the KCNK3 g...
Joseph F Cotten - One of the best experts on this subject based on the ideXlab platform.
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TASK-1 (KCNK3) and TASK-3 (KCNK9) tandem pore potassium channel antagonists stimulate breathing in isoflurane-anesthetized rats.
Anesthesia and analgesia, 2013Co-Authors: Joseph F CottenAbstract:BACKGROUND: TASK-1 and TASK-3 tandem pore potassium channel subunits provide a constitutive acidic pH- and hypoxia-inhibited potassium conductance. TASK channels are expressed in a number of tissues involved in regulation of breathing, and the TASK-1/TASK-3 heterodimer provides the predominant hypoxia-sensitive potassium conductance in carotid body type 1 glomus chemosensing cells. The carotid bodies have an important role in regulation of breathing. Doxapram is a potent TASK-1 and TASK-3 potassium channel antagonist and a carotid body and breathing stimulant. PK-THPP and A1899 are potent and selective TASK-1 and TASK-3 antagonists. I hypothesized PK-THPP and A1899 are, like doxapram, breathing stimulants. METHODS: I studied rat TASK-3 potassium channel function by Ussing chamber using Fischer rat thyroid monolayers. To quantify breathing effects, I studied male Sprague–Dawley rats spontaneously breathing 1.5% isoflurane in room air by noninvasive plethysmography and by arterial blood gas analysis. RESULTS: PK-THPP, A1899, and doxapram inhibit rat TASK-3 potassium channel function with IC50s of 42 nM (33–52), 1.6 μM (0.8–3.3), and 22 μM (18–28) (n = 4–6; 95% confidence limits). IV PK-THPP, A1899, and doxapram stimulated breathing by plethysmography with a peak change in minute ventilation relative to baseline of 84% ± 19% and 226% ± 56% (for PK-THPP at 0.5 and 5 mg/kg; mean ± SEM; n = 3–4; P 0.05 and P 0.05 for both), and 7.38 ± 0.03 and 48 ± 4 mm Hg (for dimethylsulfoxide vehicle after 1 mL/kg; n = 3). CONCLUSIONS: PK-THPP and A1899 are potent rat TASK-3 antagonists and effective breathing stimulants. PK-THPP and A1899 effects on breathing were of greater magnitude and/or duration relative to that of doxapram. PK-THPP and A1899 or related compounds may have therapeutic potential for treating breathing disorders.
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the ventilatory stimulant doxapram inhibits task tandem pore k2p potassium channel function but does not affect minimum alveolar anesthetic concentration
Anesthesia & Analgesia, 2006Co-Authors: Joseph F Cotten, Bharat Keshavaprasad, Michael J. Laster, Edmond I. Eger, Susan Connolly YostAbstract:TWIK-related acid-sensitive K + -1 (TASK-1 [KCNK3]) and TASK-3 (KCNK9) are tandem pore (K 2P ) potassium (K) channel subunits expressed in carotid bodies and the brainstem. Acidic pH values and hypoxia inhibit TASK-1 and TASK-3 channel function, and halothane enhances this function. These channels have putative roles in ventilatory regulation and volatile anesthetic mechanisms. Doxapram stimulates ventilation through an effect on carotid bodies, and we hypothesized that stimulation might result from inhibition of TASK-1 or TASK-3 K channel function. To address this, we expressed TASK-1, TASK-3, TASK-1/TASK-3 heterodimeric, and TASK-1/TASK-3 chimeric K channels in Xenopus oocytes and studied the effects of doxapram on their function. Doxapram inhibited TASK-1 (half-maximal effective concentration [EC 50 ], 410 nM), TASK-3 (ECso, 37 μM), and TASK-1/TASK-3 heterodimeric channel function (ECso, 9 μM). Chimera studies suggested that the carboxy terminus of TASK-1 is important for doxapram inhibition. Other K 2P channels required significantly larger concentrations for inhibition. To test the role of TASK-1 and TASK-3 in halothane-induced immobility, the minimum alveolar anesthetic concentration for halothane was determined and found unchanged in rats receiving doxapram by IV infusion. Our data indicate that TASK-1 and TASK-3 do not play a role in mediating the immobility produced by halothane, although they are plausible molecular targets for the ventilatory effects of doxapram.
Véronique Capuano - One of the best experts on this subject based on the ideXlab platform.
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KCNK3 Dysfunction Exaggerates The Development Of Pulmonary Hypertension Induced By Left Ventricular Pressure Overload.
Cardiovascular research, 2021Co-Authors: Mélanie Lambert, Catherine Rucker-martin, P. Mendes-ferreira, Maria-rosa Ghigna, Hélène Leribeuz, Rui Adão, Angèle Boet, Véronique Capuano, Carmen Brás-silva, Rozenn QuarckAbstract:Aims Pulmonary hypertension (PH) is a common complication of left heart disease (LHD, group 2 PH) leading to right ventricular (RV) failure and death. Several loss-of-function (LOF) mutations in KCNK3 were identified in pulmonary arterial hypertension (PAH, group 1 PH). Additionally, we found that KCNK3 dysfunction is a hallmark of PAH at pulmonary vascular and RV levels. However, the role of KCNK3 in the pathobiology of PH due to LHD is unknown. Methods and results We evaluated the role of KCNK3 on PH induced by ascending aortic constriction (AAC), in WT and KCNK3-LOF-mutated rats, by echocardiography, RV catheterization, histology analyses, and molecular biology experiments. We found that KCNK3-LOF-mutation had no consequence on the development of left ventricular (LV) compensated concentric hypertrophy in AAC, while left atrial (LA) emptying fraction was impaired in AAC-KCNK3-mutated rats. AAC-animals (WT and KCNK3-mutated rats) developed PH secondary to AAC and KCNK3-mutated rats developed more severe PH than WT. AAC-KCNK3-mutated rats developed RV and LV fibrosis in association with an increase of Col1a1 mRNA in RV and LV. AAC-KCNK3-mutated rats developed severe pulmonary vascular (pulmonary artery as well as pulmonary veins) remodelling with intense peri-vascular and peri-bronchial inflammation, perivascular edema, alveolar wall thickening, and exaggerated lung vascular cell proliferation compared to AAC-WT-rats. Finally, in lung, RV, LV, and LA of AAC-KCNK3-mutated rats, we found a strong increased expression of Il-6 and periostin expression and a reduction of lung Ctnnd1 mRNA (coding for p120 catenin), contributing to the exaggerated pulmonary and heart remodelling and pulmonary vascular edema in AAC-KCNK3-mutated rats. Conclusions Our results indicate that KCNK3-LOF is a key event in the pathobiology of PH due to AAC, suggesting that KCNK3 channel dysfunction could play a potential key role in the development of PH due to LHD.
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Implication of Potassium Channels in the Pathophysiology of Pulmonary Arterial Hypertension
Biomolecules, 2020Co-Authors: Hélène Le Ribeuz, Barbara Girerd, David Montani, Marc Humbert, Véronique Capuano, Fabrice AntignyAbstract:Pulmonary arterial hypertension (PAH) is a rare and severe cardiopulmonary disease without curative treatments. PAH is a multifactorial disease that involves genetic predisposition, epigenetic factors, and environmental factors (drugs, toxins, viruses, hypoxia, and inflammation), which contribute to the initiation or development of irreversible remodeling of the pulmonary vessels. The recent identification of loss-of-function mutations in KCNK3 (KCNK3 or TASK-1) and ABCC8 (SUR1), or gain-of-function mutations in ABCC9 (SUR2), as well as polymorphisms in KCNA5 (Kv1.5), which encode two potassium (K+) channels and two K+ channel regulatory subunits, has revived the interest of ion channels in PAH. This review focuses on KCNK3, SUR1, SUR2, and Kv1.5 channels in pulmonary vasculature and discusses their pathophysiological contribution to and therapeutic potential in PAH.
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Characterization of KCNK3 -Mutated Rat, a Novel Model of Pulmonary Hypertension
Circulation Research, 2019Co-Authors: Mélanie Lambert, Christine Péchoux, Angèle Boet, Véronique Capuano, Laurent Tesson, Thomas Bertero, Morad Nakhleh, Séverine Remy, Ignacio Anegon, Aurélie HautefortAbstract:RATIONALE: Pulmonary arterial hypertension is a severe lethal cardiopulmonary disease. Loss of function mutations in KCNK3 (potassium channel subfamily K member 3) gene, which encodes an outward rectifier K + channel, have been identified in pulmonary arterial hypertension patients. OBJECTIVE: We have demonstrated that KCNK3 dysfunction is common to heritable and nonheritable pulmonary arterial hypertension and to experimental pulmonary hypertension (PH). Finally, KCNK3 is not functional in mouse pulmonary vasculature. METHODS AND RESULTS: Using CRISPR/Cas9 technology, we generated a 94 bp out of frame deletion in exon 1 of KCNK3 gene and characterized these rats at the electrophysiological, echocardiographic, hemodynamic, morphological, cellular, and molecular levels to decipher the cellular mechanisms associated with loss of KCNK3. Using patch-clamp technique, we validated our transgenic strategy by demonstrating the absence of KCNK3 current in freshly isolated pulmonary arterial smooth muscle cells from KCNK3-mutated rats. At 4 months of age, echocardiographic parameters revealed shortening of the pulmonary artery acceleration time associated with elevation of the right ventricular systolic pressure. KCNK3-mutated rats developed more severe PH than wild-type rats after monocrotaline exposure or chronic hypoxia exposure. KCNK3-mutation induced a lung distal neomuscularization and perivascular extracellular matrix activation. Lungs of KCNK3-mutated rats were characterized by overactivation of ERK1/2 (extracellular signal-regulated kinase1-/2), AKT (protein kinase B), SRC, and overexpression of HIF1-α (hypoxia-inducible factor-1 α), survivin, and VWF (Von Willebrand factor). Linked with plasma membrane depolarization, reduced endothelial-NOS expression and desensitization of endothelial-derived hyperpolarizing factor, KCNK3-mutated rats presented predisposition to vasoconstriction of pulmonary arteries and a severe loss of sildenafil-induced pulmonary arteries relaxation. Moreover, we showed strong alteration of right ventricular cardiomyocyte excitability. Finally, KCNK3-mutated rats developed age-dependent PH associated with low serum-albumin concentration. CONCLUSIONS: We established the first KCNK3-mutated rat model of PH. Our results confirm that KCNK3 loss of function is a key event in pulmonary arterial hypertension pathogenesis. This model presents new opportunities for understanding the initiating mechanisms of PH and testing biologically relevant therapeutic molecules in the context of PH. VISUAL OVERVIEW: An online visual overview is available for this article. P ulmonary arterial hypertension (PAH) is an uncommon , progressive, and severe disease with an estimated prevalence of 15 to 50 per million. 1 PAH has been hemodynamically defined by an elevation of the mean pulmonary artery pressure >20 mm Hg and pulmonary vascular resistance >3 Wood units at rest. 2 PAH results from increased pulmonary vascular resistance because of remodeling of the small distal pulmonary arteries (PAs) and arterioles (diameter
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Characterization of KCNK3-Mutated Rat, a Novel Model of Pulmonary Hypertension.
Circulation research, 2019Co-Authors: Mélanie Lambert, Christine Péchoux, Angèle Boet, Véronique Capuano, Laurent Tesson, Thomas Bertero, Morad Nakhleh, Séverine Remy, Ignacio Anegon, Aurélie HautefortAbstract:Rationale: Pulmonary arterial hypertension is a severe lethal cardiopulmonary disease. Loss of function mutations in KCNK3 (potassium channel subfamily K member 3) gene, which encodes an outward re...
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loss of KCNK3 is a hallmark of rv hypertrophy dysfunction associated with pulmonary hypertension
Cardiovascular Research, 2018Co-Authors: Mélanie Lambert, Catherine Ruckermartin, Angèle Boet, Véronique Capuano, Stéphane N. Hatem, P Mendesferreira, R AdaoAbstract:Aims Mutations in the KCNK3 gene, which encodes for an outward-rectifier K+ channel, have been identified in patients suffering from pulmonary arterial hypertension (PAH), and constitute the first described channelopathy in PAH. In human PAH and experimental pulmonary hypertension (PH), we demonstrated that KCNK3 expression and function are severely reduced in pulmonary vascular cells, promoting PH-like phenotype at the morphologic and haemodynamic levels. Since KCNK3 channel is also expressed in both the human and rodent heart, we aimed to elucidate the pathophysiological role of KCNK3 channel in right ventricular (RV) hypertrophy (RVH) related to PH. Methods and results Using whole-cell Patch-clamp technique, we demonstrated that KCNK3 is predominantly expressed in adult rat RV cardiomyocytes compared to the left ventricle cardiomyocytes and participates in the repolarizing phase of the RV action potential. We revealed a reduction in KCNK3 function prior to development of RVH and the rise of pulmonary vascular resistance. KCNK3 function is severely reduced in RV cardiomyocytes during the development of RVH in several rat models of PH (exposure to monocrotaline, chronic hypoxia, and Sugen/hypoxia) and chronic RV pressure overload (pulmonary artery banding). In experimental PH, we revealed a reduction in KCNK3 function before any rise in pulmonary vascular resistance and the development of RVH. KCNK3 mRNA level is also reduced in human RV tissues from PAH patients compared to non-PAH patients. In line with these findings, chronic inhibition of KCNK3 in rats with the specific inhibitor (A293) induces RV hypertrophy which is associated with the re-expression of foetal genes, RV fibrosis, RV inflammation, and subsequent loss of RV performance as assessed by echocardiography. Conclusion Our data indicate that loss of KCNK3 function and expression is a hallmark of the RV hypertrophy/dysfunction associated with PH.