Progressive degenerative role of Nox and thrombospondin-1 in the aging vasculature

Nox 和血小板反应蛋白-1 在衰老脉管系统中的进行性退行性作用

• 批准号: 10115104
• 负责人: Patrick J Pagano
• 金额: $ 57.71万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10115104
• 关键词: 3-Dimensional; Age; Aging; Agonist; Animals; Arteries; Attenuated; Binding; Blocking Antibodies; Blood; Blood Vessels; Blood flow; CD47 gene; Cardiovascular Diseases; Cell Aging; Cell membrane; Clinical Data; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Development; Elderly; Endothelial Cells; Endothelium; GKLF protein; Genes; Genomics; Hindlimb; Human; Hypertension; Impairment; In Vitro; Individual; Intercellular Fluid; Ischemia; Knock-out; Knockout Mice; Lasers; Lead; Ligation; Link; MYC gene; Mediating; Mesentery; Molecular Genetics; Molecular Target; Mus; Myocardial Infarction; NADPH Oxidase 1; Oxidants; PF4 Gene; Pathologic; Patients; Perfusion; Peripheral; Peripheral Vascular Diseases; Pharmacology; Plasma; Production; Rattus; Reactive Oxygen Species; Reporting; Resistance; Risk Factors; Role; Severity of illness; Signal Pathway; Stroke; TP53 gene; Testing; Thrombospondin 1; Tissues; Triad Acrylic Resin; Tube; Vascular Diseases; Vasodilator Agents; Vasomotor; Wild Type Mouse; age related; aged; angiogenesis; c-myc Genes; effective therapy; endothelial dysfunction; femoral artery; healing; human tissue; improved; in vitro testing; in vivo; in vivo evaluation; inhibitor/antagonist; interstitial; loss of function; microangiography; middle age; new therapeutic target; novel; peripheral blood vessel; pre-clinical; receptor; response; self-renewal; senescence; sex; therapeutic target; tool; transcription factor; vascular stress

PROJECT SUMMARY Aging is a recognized risk factor in peripheral vascular disease (PVD). A hallmark of aging in humans is loss of vascular function reflected, in part, by inhibited self-renewal of endothelial cells (EC). An unmet need is the emergence of therapies that stop or reverse endothelial senescence (decreased capacity for cellular division, self-renewal) and vascular insufficiency. A second pathognomonic feature of aging is increased reactive oxygen species (ROS). Pathologic ROS impair vascular flow, but the mechanisms responsible for these maladaptive age-related changes in the endothelium remain unknown. Thrombospondin-1 (TSP1) is increased in the plasma, arterial walls and interstitial fluid of peripheral vascular disease (PVD) patients and is linked to disease severity and loss of vascularity. We have found that with aging, wild type (WT) mice show induction of TSP1 and its cognate cell membrane receptor CD47 and this was associated with decreased vasodilator- mediated changes in blood flow, an effect not observed in TSP1-/- and CD47-/- mice. We also reported that TSP1, via CD47, potently stimulates NADPH oxidase 1 (Nox1)-derived ROS and impedes hind-limb blood flow. Preliminary data by our group show that TSP1 is upregulated in older adults, and it inhibits endothelial self- replication and promotes endothelial senescence in peripheral vessels. These findings led us to propose the overarching hypothesis that TSP1, via CD47 engagement and Nox1 activation, impairs self-renewal through exacerbated ROS production culminating in EC senescence and vasculopathy. Three specific aims will be pursued to: (1) test in vitro that with aging TSP1 induces Nox1-mediated suppression of key self- renewal genes and promotes endothelial cell (EC) senescence; (2) explore in vivo that inhibition of the TSP1-CD47 axis as well as attenuating its downstream Nox1 effector restores self-renewal, angiogenesis and perfusion in aged animals; and (3) investigate that perturbing the TSP1-CD47-Nox1 axis in aged human tissue ameliorates age-related endothelial and vascular dysfunction. The proposal will employ multiple molecular and genetic tools to interrogate mechanisms into TSP1-CD47-mediated activation of Nox1 in aging human and mouse vessel function, self-renewal and angiogenesis, as well as generate two EC-specific knockout mice to test our hypotheses. These studies will also provide robust pre- clinical data towards rational development of new and effective treatments for EC senescence and vasculopathy.

项目概要 衰老是外周血管疾病（PVD）的公认危险因素。人类衰老的一个标志是丧失 血管功能部分反映为内皮细胞（EC）自我更新受到抑制。未满足的需求是 阻止或逆转内皮衰老（细胞分裂能力下降， 自我更新）和血管供血不足。衰老的第二个特征是反应性增加 氧物质（ROS）。病理性 ROS 会损害血管血流，但其机制 与年龄相关的内皮适应不良变化仍然未知。血小板反应蛋白-1 (TSP1) 增加 存在于周围血管疾病 (PVD) 患者的血浆、动脉壁和间质液中，并且与 疾病的严重程度和血管的丧失。我们发现，随着衰老，野生型 (WT) 小鼠表现出诱导 TSP1 及其同源细胞膜受体 CD47，这与血管舒张剂减少有关 介导血流变化，但在 TSP1-/- 和 CD47-/- 小鼠中未观察到这种效应。我们还报道了 TSP1 通过 CD47 有效刺激 NADPH 氧化酶 1 (Nox1) 衍生的 ROS 并阻碍后肢血流。 我们小组的初步数据表明，TSP1 在老年人中表达上调，并且抑制内皮细胞自身 复制并促进外周血管内皮衰老。这些发现促使我们提出 总体假设是，TSP1 通过 CD47 参与和 Nox1 激活，通过以下方式损害自我更新： ROS 产生加剧，最终导致 EC 衰老和血管病变。将实现三个具体目标 目的是：（1）体外测试随着老化 TSP1 诱导 Nox1 介导的关键自我抑制 更新基因并促进内皮细胞（EC）衰老； (2) 探索体内抑制 TSP1-CD47 轴以及减弱其下游 Nox1 效应器恢复自我更新， 老年动物的血管生成和灌注； (3) 研究干扰 TSP1-CD47-Nox1 衰老人体组织中的轴可改善与年龄相关的内皮和血管功能障碍。提案 将采用多种分子和遗传工具来探究 TSP1-CD47 介导的机制 Nox1 的激活对人类和小鼠衰老血管功能、自我更新和血管生成以及 生成两只 EC 特异性基因敲除小鼠来测试我们的假设。这些研究还将提供强有力的预 合理开发新的、有效的 EC 衰老治疗方法的临床数据 血管病变。