A ROLE FOR EYA3 IN VASCULAR REMODELING AND PULMONARY ARTERIAL HYPERTENSION

EYA3 在血管重塑和肺动脉高压中的作用

• 批准号: 10171900
• 负责人: RASHMI S. HEGDE
• 金额: $ 63.62万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10171900
• 关键词: Animal Model; Animals; Apoptosis; Biochemical; Biological Models; Blood Vessels; Cell Survival; Cells; Cessation of life; Characteristics; Chronic; DNA Damage; DNA Repair; Data; Development; Diagnosis; Disease; Disease model; Distal; Drug Design; Drug Targeting; Effectiveness; Endothelium; Enzymes; Evaluation; Event; Experimental Models; Eye; Functional disorder; Genetic; Genetic Transcription; Goals; Heart failure; Human; Hypoxia; In Vitro; Inflammation; Intervention; Investigation; Lead; Life; Lung; Maintenance; Modeling; Molecular; Mus; Muscle; Mutation; Oxidative Stress; Pathogenesis; Pathologic; Pathology; Pathway interactions; Patients; Pharmacology; Phenotype; Phosphoproteins; Phosphoric Monoester Hydrolases; Protein Tyrosine Phosphatase; Proteins; Protocols documentation; Pulmonary Hypertension; Pulmonary Vascular Resistance; Pulmonary artery structure; Rattus; Resistance; Role; Safety; Series; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Specificity; Structure; Symptoms; Testing; Therapeutic; Therapeutic Effect; Tissues; Transactivation; Transcription Coactivator; Transgenic Mice; Validation; Vascular Endothelium; Vascular remodeling; Vascular resistance; arteriole; base; cell injury; cell type; defined contribution; hemodynamics; in vivo; inhibitor/antagonist; insight; link protein; mouse model; novel; novel therapeutic intervention; pharmacophore; phosphatase inhibitor; pre-clinical; premature; pressure; prevent; protective effect; pulmonary arterial hypertension; repaired; single-cell RNA sequencing; therapeutic target; transcriptomics; treatment strategy

Project Summary Pulmonary Hypertension (PH) is a pathophysiologic condition characterized by elevated pressure in the pulmonary arteries. Pulmonary arterial hypertension (WHO Group I PH; PAH) is a particularly severe form of PH frequently associated with right heart failure and premature death. There is no cure, and treatments only target the symptoms. Approximately 50% of PAH patients die within 5 years of diagnosis. There is therefore a compelling, unmet need for new therapeutic strategies. Pulmonary vascular remodeling is the defining pathological feature of PAH. It leads to occlusion of distal pulmonary arterioles, with accompanying increase in pulmonary vascular resistance. Vascular remodeling is promoted by the survival and proliferation of pulmonary arterial vascular cells under conditions of oxidative stress and in the presence of DNA damage. Here we provide evidence that the Eyes Absent protein (EYA3) promotes survival of DNA damaged cells facing a survival-versus-apoptosis decision. EYA3 is a druggable and mechanistically unique protein tyrosine phosphatase (PTP) present at elevated levels in pulmonary arterial smooth muscle cells isolated from PAH patients. In proof of principle studies, we show that transgenic mice harboring an inactivating mutation in the EYA3 PTP domain are significantly protected from vascular remodeling in a chronic hypoxia model, and that inhibitors of EYA3 PTP activity reverse vascular remodeling in a rat model of experimental angio-obliterative PH. The overall goal of this project is to establish the EYA3-PTP as a disease-modifying target whose function in the pathophysiology of PAH can be targeted by available inhibitors. This will be a critical milestone in pre-clinical drug target validation and will be achieved through the use of genetic and pharmacological approaches in the following two Specific Aims: Aim I: To elucidate the molecular mechanism(s) through which EYA3 promotes the pathogenesis of PAH we will use a murine chronic hypoxia model. Hemodynamic and histopathological analyses upon cell-type specifc EYA3 deletion will be conducted to identify the cell-autuonomous roles of EYA3 in vascular remodeling. Single-cell transcriptomics and phospho-protein analyses will be used to delineate the molecular mechanims contributing to vascular remodeling. Aim II: To establish the effectiveness of EYA3-PTP inhibitors in reversing established vascular remodeling we will use a rat model of angio-obliterative PAH and evaluate the most safe and effective modes of EYA3-targeted pharmacological intervention. The overall impact of this proposal is that it will define a targetable signaling mechanism that contributes to the characteristic pulmonary vascular pathobiology in PAH, demonstrate that EYA3-PTP inhibitors are viable lead compounds for the development of PAH therapeutics, and provide insights into a previously unexplored molecular mechanism contributing to PAH pathology.

项目摘要 肺动脉高压（pH）是一种病理生理状况，其特征是升高 肺动脉的压力。肺动脉高压（谁是I组； PAH）是一种特别严重的pH形式，经常与正确的心力衰竭相关 过早死亡。无法治愈，治疗只针对症状。 大约50％的PAH患者在诊断后的5年内死亡。因此有一个 引人注目的，未满足的对新的治疗策略的需求。 肺血管重塑是PAH的定义病理特征。它导致 远端肺动脉的阻塞，伴有肺部增加 血管抗性。血管重塑是通过生存和增殖来促进的 在氧化应激条件下，肺动脉血管细胞和存在 DNA损伤。在这里，我们提供的证据表明眼睛没有蛋白质（EYA3） 促进DNA损坏的细胞的存活，面临生存 - 凋亡决定。 EYA3是一种可吸毒且机械独特的蛋白酪氨酸磷酸酶（PTP） 从PAH分离 患者。在原则研究的证明中，我们表明具有携带的转基因小鼠 EYA3 PTP结构域中的灭活突变受到显着保护 在慢性缺氧模型中进行重塑，EYA3 PTP活性的抑制剂反向 在实验性血管生成pH的大鼠模型中进行血管重塑。 该项目的总体目标是将EYA3-PTP建立为疾病改良 可以通过可用的针对PAH病理生理学的功能的目标 抑制剂。这将是临床前药物目标验证的关键里程碑，将是 通过使用遗传和药理学方法在以下 两个具体的目标： 目的I：阐明EYA3促进的分子机制 PAH的发病机理我们将使用鼠慢性低氧模型。血流动力和 将对细胞类型的EYA3缺失进行组织病理学分析 确定EYA3在血管重塑中的细胞自动作用。单细胞 转录组学和磷酸蛋白分析将用于描述分子 有助于血管重塑的机械。 AIM II：建立EYA3-PTP抑制剂在逆转建立的有效性 血管重塑，我们将使用符合血管生成的大鼠模型，并评估 最安全有效的EYA3靶向药理学干预模式。 该提案的总体影响是它将定义可靶向信号 有助于特征性肺血管病理学的机制 PAH，证明EYA3-PTP抑制剂是可行的铅化合物 开发PAH疗法，并提供有关先前未开发的见解 有助于PAH病理学的分子机制。