Role of Endothelial Anoctamin-1 in Pulmonary Arterial Hypertension

内皮 Anoctamin-1 在肺动脉高压中的作用

基本信息

批准号：
9803089
负责人：
Gaurav Choudhary
金额：
$ 43.08万
依托单位：
OCEAN STATE RESEARCH INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9803089
关键词：
Animal Model Animals Apoptosis Apoptotic Attenuated Blood Vessels Calcium Cell Line Cell Proliferation Cell Proliferation Regulation Cell membrane Cells Chloride Channels Chlorides Conflict (Psychology)DNA DNA Methylation Disease Dissociation Endothelial Cells Endothelium Epidermal Growth Factor Receptor Epigenetic Process Foundations Growth In Vitro Inhalation Ion Channel Lesion Lung Malignant Neoplasms Medial Mediating Methylation Mitochondria Morbidity - disease rate OPA1 gene Optic Atrophy Outcome Pathology Patients Pharmacology Phenotype Pre-Clinical Model Proteins Pulmonary Hypertension Pulmonary artery structure Rattus Regulatory Element Resistance Role Signal Pathway Signal Transduction Site Testing Therapeutic Tissues Vascular remodeling design effective therapy improved in vitro testing in vivo methylation pattern mortality novel novel therapeutics overexpression preclinical study pressure prevent pulmonary arterial hypertension response small molecule treatment strategy vasoconstriction

项目摘要

ABSTRACT Pulmonary arterial hypertension (PAH) is associated with significant morbidity and mortality. PAH pathology includes vasoconstriction, medial and adventitial remodeling, and microvascular endothelial cell (EC) proliferation leading to vaso-occlusive plexiform lesions. EC in PAH are both apoptosis resistant and hyperproliferative. There are no effective treatments for the severe vascular remodeling observed with PAH, and therapeutic strategies that target the dysfunctional EC and plexiform lesions are urgently needed. We have discovered that calcium activated chloride channel, Ano1, is localized to both the EC plasma membrane (pl-Ano1) and mitochondria (mito-Ano1). Ano1 expression is upregulated in EC in settings of PAH and associated with a hyperproliferative and apoptosis resistant phenotype. However, unregulated activation of this channel results in apoptosis of hyperproliferative apoptosis-resistant ECs. Our overall objective in this proposal is to delineate the mechanisms underlying these seemingly conflicting observations to identify therapeutic opportunities to decrease EC proliferation and target hyperproliferative ECs for apoptosis, thereby improving pulmonary vascular remodeling in PAH. We hypothesize that epigenetically regulated increased expression of Ano1 results in EC proliferation and apoptosis resistance, yet unregulated opening of Ano1 in the context of overexpression is detrimental and can potentially be exploited to promote apoptosis of hyperproliferative ECs in PAH. In Aim 1, we will determine the mechanism of increased Ano1 expression in PAH EC with a focus on DNA methylation. We will use complementary in vitro and in vivo approaches to design novel constructs to selectively alter the methylation pattern of Ano1 regulatory elements in lung EC and test if this strategy prevents or treats PAH. In Aim 2, we will determine the effect of increased pl- and mito- Ano1 expression on EC proliferation and apoptosis resistance and elucidate the underlying mechanisms. We will investigate the signaling pathways in vitro and then use selective inhibition of Ano1 expression in lung EC in vivo to determine its effect on preventing or treating PAH. In Aim 3, we will determine the mechanism of the effect of pl- and mito-Ano-1 opening on EC apoptosis in vitro and test if opening of Ano1 by a small molecule delivered by inhalation attenuates PAH. As a result of these studies we expect to delineate the mechanism of Ano1-mediated EC proliferation and apoptosis resistance in settings of PAH as well as develop novel therapeutic strategy targeting Ano1 to improve pulmonary vascular remodeling and PAH.

抽象的肺动脉高压（PAH）与显着的发病率和死亡率相关。多环芳烃病理学包括血管收缩、内侧和外膜重塑以及微血管内皮细胞 (EC) 增殖导致血管闭塞丛状病变。 PAH 中的 EC 既具有细胞凋亡抗性，过度增殖。对于 PAH 所观察到的严重血管重塑，尚无有效的治疗方法，迫切需要针对功能失调的 EC 和丛状病变的治疗策略。我们发现钙激活的氯离子通道 Ano1 定位于 EC 质膜 (pl-Ano1) 和线粒体 (mito-Ano1)。在 PAH 和环境下 EC 中 Ano1 表达上调与过度增殖和抗凋亡表型相关。然而，这种不受控制的激活通道导致过度增殖的抗凋亡 EC 的凋亡。我们在此方面的总体目标建议是描述这些看似相互矛盾的观察结果背后的机制，以确定减少 EC 增殖并针对过度增殖的 EC 进行凋亡的治疗机会，从而改善 PAH 的肺血管重塑。我们假设表观遗传调控增加了 Ano1 的表达导致 EC 增殖和细胞凋亡抵抗，但 Ano1 在细胞中不受调控的开放过度表达的情况是有害的，并且可能被用来促进细胞凋亡 PAH 中过度增殖的 EC。在目标 1 中，我们将确定 Ano1 表达增加的机制 PAH EC 重点关注 DNA 甲基化。我们将使用互补的体外和体内方法设计新颖的结构来选择性改变肺 EC 中 Ano1 调节元件的甲基化模式测试该策略是否可以预防或治疗 PAH。在目标 2 中，我们将确定增加 pl- 和 mito- 的效果 Ano1 表达对 EC 增殖和凋亡抵抗的影响并阐明其潜在机制。我们将研究体外信号通路，然后选择性抑制肺 EC 中的 Ano1 表达体内测定其预防或治疗 PAH 的效果。在目标 3 中，我们将确定 pl-和mito-Ano-1开放对体外EC细胞凋亡的影响并测试小分子是否开放Ano1 通过吸入传递可减弱 PAH。作为这些研究的结果，我们希望能够描述出其机制 PAH 环境中 Ano1 介导的 EC 增殖和细胞凋亡抵抗以及开发新的针对 Ano1 改善肺血管重塑和 PAH 的治疗策略。