ConProject-001

ConProject-001

• 批准号: 10204790
• 负责人: Ashish Kumar Sharma
• 金额: $ 58.39万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-09 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10204790
• 关键词: ATP Receptors; ATP phosphohydrolase; Abdomen; Abdominal Aortic Aneurysm; Adhesions; Aneurysm; Angiography; Animals; Aortic Aneurysm; Aortic Rupture; Aortitis; Apolipoprotein E; Apyrase; Attenuated; Autocrine Communication; Balloon Angioplasty; Biological; Blood Platelets; Bone Marrow; Caliber; Catabolism; Cells; Chronic; Clinical; Clinical Trials; Coculture Techniques; Collagen; Data; Elastases; Elastin; Elderly; Emigrations; Endothelial Cells; Endothelium; Enzymes; Erythrocytes; Exposure to; Family suidae; Future; Gelatinase A; Genetic; Goals; HMGB1 gene; Human; IL2 gene; Immune; In Vitro; Infiltration; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Interleukin-1 beta; Interleukin-17; Knock-out; Knockout Mice; Lead; Leukocytes; Macrophage Activation; Measures; Mediating; Mediator of activation protein; Medical; Modality; Modeling; Molecular; Mus; Nucleotides; P2X-receptor; P2Y2 receptor; Pathogenesis; Patients; Peptides; Perfusion; Pharmacology; Phenotype; Population; Prevention; Probenecid; Production; Purinoceptor; Reporting; Reproducibility; Research Personnel; Risk; Role; Rupture; Signal Transduction; Smooth Muscle Actin Staining Method; Smooth Muscle Myocytes; Source; Sudden Death; TNF gene; Testing; Therapeutic; Thinness; Tissue Inhibitor of Metalloproteinases; Tissues; Tunica Adventitia; Ultrasonography; Vascular Endothelial Cell; Vascular remodeling; base; beta Globin; clinically relevant; cytokine; ectoATPase; efficacy evaluation; experimental study; extracellular; high risk; immune activation; inhibitor/antagonist; innovation; insight; knockout animal; leukocyte activation; macrophage; migration; mortality; mouse model; neutrophil; new therapeutic target; novel; porcine model; pre-clinical; receptor; reconstitution; response; therapeutic target; therapeutically effective; tissue injury; trovafloxacin; vascular inflammation

Project Summary Aortic aneurysms (AA) represent a significant clinical problem with a high risk of aortic rupture leading to sudden death. Currently, there are no medical therapies available to treat aneurysms and a clear understanding of the pathobiology of AA formation is key to develop therapeutic pharmacological approaches. Recent studies reveal that ATP can be released by cells in a controlled manner through pannexin 1 (Panx1) channels to signal through purinergic P2X or P2Y receptors. ATP is a nucleotide released in large amounts after tissue injury such as AA and serves as a “danger signal” to mediate inflammation. Our data suggest that Panx1 on endothelial cells (ECs) is an important mediator of vascular inflammation in a murine model of abdominal AA (AAA) and can be a major source of extracellular ATP. The expression of Panx1 is highest in the arterial network especially in the aortic endothelium. The scientific premise of this proposal focuses on the crosstalk between aortic endothelium, smooth muscle cells and relevant immune cells to delineate Panx1- mediated signaling in AAA progression. Thus, this proposal will test the overall hypothesis that EC Panx1 signaling is a major, early inflammatory mediator of the signaling cascade via release of ATP resulting in smooth muscle cell activation, leukocyte infiltration and vascular inflammation. Using novel inducible cell- specific genetic knockout animals, specific pharmacological Panx1 inhibitors and P2X/P2Y antagonists, we will delineate this previously unknown mechanism of AAA pathobiology. Aim 1 will determine if EC Panx1 signaling mediates AAA via ATP release and its crosstalk with aortic smooth muscle cells as well as to determine whether the P2X7 receptor is a major determinant of AAA. We will also investigate if Panx1 inhibition can treat preformed AAAs in the murine models. Aim 2 will determine if Panx1 signaling mediates leukocyte activation and infiltration during AAA. The role of Panx1 and specific purinergic receptors on macrophages will be deciphered as well as whether Panx1 mediates neutrophil transendothelial migration. Aim 3 will determine if pharmacologic inhibition of Panx1 or specific P2 receptors will treat AAA using a chronic inflammatory, large animal porcine AAA model. Our studies will provide novel insight into mechanisms of molecular signaling interactions between aortic endothelial cells, smooth muscle cells, and immune cells (macrophages and neutrophils), to define Panx1 as a novel therapeutic target for the treatment of AAs.

项目摘要 主动脉瘤（AA）是一个重要的临床问题，具有高风险的主动脉破裂，导致 突然死亡目前，没有可用的医学疗法来治疗动脉瘤， 对AA形成的病理生物学的理解是开发治疗药理学方法的关键。 最近的研究表明，ATP可以通过泛连接蛋白1（Panx 1）以受控的方式由细胞释放。 通过嘌呤能P2 X或P2 Y受体传递信号。ATP是一种大量释放的核苷酸 在组织损伤如AA后，作为“危险信号”介导炎症。我们的数据表明 内皮细胞上的Panx 1是血管炎小鼠模型中血管炎症的重要介质， 腹AA（AAA），可以是细胞外ATP的主要来源。Panx 1的表达最高， 动脉网络，尤其是主动脉内皮。这一建议的科学前提侧重于 主动脉内皮、平滑肌细胞和相关免疫细胞之间的串扰，以描绘Panx 1- 在AAA进展中介导的信号传导。因此，本提案将检验EC Panx 1 信号传导是信号传导级联的主要的早期炎症介质，通过释放ATP， 平滑肌细胞活化、白细胞浸润和血管炎症。使用新的可诱导细胞- 特定的基因敲除动物，特定的药理学Panx 1抑制剂和P2 X/P2 Y拮抗剂，我们将 描述了AAA病理生物学的这一先前未知的机制。目标1将确定EC Panx 1是否 信号转导通过ATP释放及其与主动脉平滑肌细胞的串扰介导AAA， 确定P2 X7受体是否是AAA的主要决定因素。我们还将调查Panx 1是否 抑制可以治疗鼠模型中预先形成的AAA。目标2将确定Panx 1信号传导是否介导 AAA期间白细胞活化和浸润。Panx 1和特异性嘌呤能受体在细胞凋亡中的作用 巨噬细胞以及Panx 1是否介导嗜中性粒细胞跨内皮迁移。 目的3将确定Panx 1或特异性P2受体的药理学抑制是否将使用慢性给药治疗AAA。 炎性大动物猪AAA模型。我们的研究将提供新的洞察机制， 主动脉内皮细胞、平滑肌细胞和免疫细胞之间的分子信号相互作用 （巨噬细胞和中性粒细胞），将Panx 1定义为治疗AA的新治疗靶点。