title: Inhibition of Excessive Cell Proliferation by Calcilytics in Idiopathic Pulmonary Arterial Hypertension.
abstract: Idiopathic pulmonary arterial hypertension (IPAH) is a rare and progressive disease of unknown pathogenesis. Vascular remodeling due to excessive proliferation of pulmonary arterial smooth muscle cells (PASMCs) is a critical pathogenic event that leads to early morbidity and mortality. The excessive cell proliferation is closely linked to the augmented Ca2+ signaling in PASMCs. More recently, we have shown by an siRNA knockdown method that the Ca2+-sensing receptor (CaSR) is upregulated in PASMCs from IPAH patients, involved in the enhanced Ca2+ response and subsequent excessive cell proliferation. In this study, we examined whether pharmacological blockade of CaSR attenuated the excessive proliferation of PASMCs from IPAH patients by MTT assay. The proliferation rate of PASMCs from IPAH patients was much higher (~1.5-fold) than that of PASMCs from normal subjects and patients with chronic thromboembolic pulmonary hypertension (CTEPH). Treatment with NPS2143, an antagonist of CaSR or calcilytic, clearly suppressed the cell proliferation in a concentration-dependent manner (IC50 = 2.64 μM) in IPAH-PASMCs, but not in normal and CTEPH PASMCs. Another calcilytic, Calhex 231, which is structurally unrelated to NPS2143, also concentration-dependently inhibited the excessive proliferation of IPAH-PASMCs (IC50 = 1.89 μM). In contrast, R568, an activator of CaSR or calcimimetic, significantly facilitated the proliferation of IPAH-PASMCs (EC50 = 0.33 μM). Similar results were obtained by BrdU incorporation assay. These results reveal that the excessive PASMC proliferation was modulated by pharmacological tools of CaSR, showing us that calcilytics are useful for a novel therapeutic approach for pulmonary arterial hypertension.
authors.affiliation: Department of Pharmacy, College of Pharmacy, Kinjo Gakuin University, Nagoya, Japan.
authors.forename: Aya
authors.initials: A
authors.lastname: Yamamura
authors.affiliation: Department of Pharmacy, College of Pharmacy, Kinjo Gakuin University, Nagoya, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Ohara
authors.affiliation: Department of Pharmacy, College of Pharmacy, Kinjo Gakuin University, Nagoya, Japan.
authors.forename: Kikuo
authors.initials: K
authors.lastname: Tsukamoto
mesh_terms: D000814:Aniline Compounds / Q000494:pharmacology; D001549:Benzamides / Q000494:pharmacology; D002118:Calcium / Q000819:agonists; D016022:Case-Control Studies; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D002908:Chronic Disease; D003514:Cyclohexylamines / Q000494:pharmacology; D065627:Familial Primary Pulmonary Hypertension / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D009281:Naphthalenes / Q000494:pharmacology; D010627:Phenethylamines; D011437:Propylamines; D011651:Pulmonary Artery / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D011655:Pulmonary Embolism / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D044169:Receptors, Calcium-Sensing / Q000037:antagonists & inhibitors*; D013923:Thromboembolism / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*
chemical_list: D000814:Aniline Compounds; D001549:Benzamides; C095550:CASR protein, human; D003514:Cyclohexylamines; C482333:N(1)-(4-chlorobenzoyl)-N(2)-(1-(1-naphthyl)ethyl)-1,2-diaminocyclohexane; C107873:N-(2-chlorophenylpropyl)-1-(3-methoxyphenyl)ethylamine; C436740:N-(2-hydroxy-3-(2-cyano-3-chlorophenoxy)propyl)-1,1-dimethyl-2-(2-nephthyl)ethylamine; D009281:Naphthalenes; D010627:Phenethylamines; D011437:Propylamines; D044169:Receptors, Calcium-Sensing; D002118:Calcium