Molecular Drivers of Vascular Stiffness and Metabolic Dysfunction in HIV-Induced Pulmonary Arterial Hypertension

HIV 引起的肺动脉高压中血管僵硬和代谢功能障碍的分子驱动因素

• 批准号: 9366038
• 负责人: Stephen Y Chan
• 金额: $ 77.35万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9366038
• 关键词: Animal Model; Animals; Aspartate; Binding; Biological Assay; Blood Vessels; Catheterization; Cells; Clinical Trials; Coculture Techniques; Complex; Data; Detection; Development; Diagnostic; Disease; Early Diagnosis; Endocrine; Endothelial Cells; Endothelium; Ensure; Extracellular Matrix; Family; Functional disorder; Glutamates; Glutaminase; Glutamine; HIV; HIV Infections; Hematological Disease; Human; In Vitro; Infection; Knockout Mice; Link; Lung; Macaca; Malignant Neoplasms; Mammals; Mediating; Metabolic; Metabolism; MicroRNAs; Modality; Modeling; Molecular; Mus; Oral; Pathogenicity; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Pharmacology; Phenotype; Plasma; Positioning Attribute; Prevalence; Primates; Process; Production; Protocols documentation; Publishing; Pulmonary Artery Catheterization; Pulmonary artery structure; Reporting; Risk; Rodent; Rodent Model; SIV; Sampling; T-Lymphocyte; Techniques; Testing; Transcription Coactivator; Transfusion; Up-Regulation; Vascular Diseases; arterial stiffness; base; hemodynamics; human disease; improved; in vivo; inhibitor/antagonist; insight; loss of function; molecular diagnostics; neglect; new therapeutic target; novel; novel diagnostics; pulmonary arterial hypertension; pulmonary artery endothelial cell; targeted treatment

Background: Pulmonary arterial hypertension (PAH) is a deadly vascular disease with increased prevalence with human immunodeficiency virus (HIV) infection. HIV-PAH may be even more prevalent than originally anticipated, yet diagnostics and treatments are limited for this enigmatic form of PAH. Recently, we have found that vascular stiffening and glutamine metabolism are linked processes in PAH, including primate and human examples of HIV-PAH. The transcriptional co-activators YAP/TAZ induce the microRNA (miR)-130/301 family and glutaminase (GLS1) to control these phenotypes. In PAH rodent models, inhibition of miR-130/301, YAP, or GLS1 improve PAH. We also found that HIV-infected T cells release cell-free miR-21 molecules and up-regulate glutaminolytic and matrix remodeling pathways in co-cultured pulmonary vascular cells. We hypothesize that a YAP/TAZ-miR-130/301-GLS1 axis is induced by a miR-21-mediated process, linking HIV-infected T cells and pulmonary vascular cells and thus activating vascular stiffening, glutaminolysis, and HIV-PAH. Specific Aims: Aim 1) Determine if miR-21 released from HIV-infected T cells induces pulmonary vascular metabolic dysfunction and matrix remodeling. In vitro, we will investigate the direct delivery and actions of miR-21 to pulmonary vascular cells from HIV-infected T cells. In vivo, employing transfusions of miR-21-replete vs. miR-21-depleted plasma into miR-21-/- mice vs. wildtype mice, we will determine if circulating miR-21 is delivered to pulmonary endothelium and induces vascular stiffening, glutaminolysis, and PAH. Results could establish an entirely novel miR link between HIV infection and pulmonary vascular glutaminolysis and stiffness. Aim 2) Determine if pulmonary arterial stiffness and glutaminolysis are evident in humans with HIV-PAH. From prior collected HIV-PAH samples, we will correlate plasma miR-21 levels with pulmonary artery (PA) compliance as calculated from hemodynamic data and plasma metabolites reflective of PA glutaminolysis. Using optimized techniques based on our published protocols, we will also assess for activation of the YAP/TAZ-miR- 130/301-GLS1 axis in PA endothelial cells collected via catheterization of HIV-PAH patients. These results could establish this mechanism in human HIV-PAH and suggest needed molecular diagnostics for HIV-PAH detection. Aim 3) Determine if up-regulation of GLS1 is necessary for promoting SIV-PAH. In SIV-PAH macaques, we will administer CB-839, an oral GLS1 inhibitor being tested in human cancer trials, to determine its effects on stiffness, glutaminolysis, and PAH. Results could demonstrate the direct pathogenic actions of GLS1 in HIV- relevant PAH and thus could re-purpose this drug for rapid, expedited trials in human HIV and PAH patients. Significance: Our team is uniquely positioned for making major molecular discoveries of HIV-PAH. We will leverage the only known reliable animal model of HIV-PAH with human studies, ensuring mechanistic insight and applicability to human disease to an extent never possible before. Perhaps most importantly, it offers a rare opportunity to establish a much needed targeted therapeutic for this historically neglected vascular disease.

背景：肺动脉高压（PAH）是一种致命的血管疾病，患病率不断增加 患有人类免疫缺陷病毒（HIV）感染。 HIV-PAH 可能比原来更普遍 预期，但对于这种神秘形式的多环芳烃的诊断和治疗是有限的。最近，我们发现 血管硬化和谷氨酰胺代谢是 PAH 中相关的过程，包括灵长类动物和人类 HIV-PAH 的例子。转录共激活因子 YAP/TAZ 诱导 microRNA (miR)-130/301 家族 和谷氨酰胺酶（GLS1）来控制这些表型。在 PAH 啮齿动物模型中，抑制 miR-130/301、YAP 或 GLS1改善PAH。我们还发现，HIV 感染的 T 细胞释放无细胞 miR-21 分子并上调 共培养的肺血管细胞中的谷氨酰胺分解和基质重塑途径。我们假设一个 YAP/TAZ-miR-130/301-GLS1 轴由 miR-21 介导的过程诱导，连接 HIV 感染的 T 细胞 和肺血管细胞，从而激活血管硬化、谷氨酰胺分解和 HIV-PAH。 具体目标： 目标 1) 确定 HIV 感染的 T 细胞释放的 miR-21 是否诱导肺血管生成 代谢功能障碍和基质重塑。在体外，我们将研究直接递送和作用 miR-21 到来自 HIV 感染 T 细胞的肺血管细胞。在体内，采用富含 miR-21 的输注 与 miR-21-/- 小鼠和野生型小鼠中的 miR-21 耗尽血浆相比，我们将确定循环 miR-21 是否 递送至肺内皮并诱导血管硬化、谷氨酰胺分解和 PAH。结果可以 在 HIV 感染与肺血管谷氨酰胺分解和僵硬之间建立了一种全新的 miR 联系。 目标 2) 确定 HIV-PAH 患者是否存在明显的肺动脉僵硬度和谷氨酰胺分解作用。 根据之前收集的 HIV-PAH 样本，我们将血浆 miR-21 水平与肺动脉 (PA) 相关联 根据血流动力学数据和反映 PA 谷氨酰胺分解的血浆代谢物计算得出的依从性。使用 基于我们发布的协议优化技术，我们还将评估 YAP/TAZ-miR- 的激活 通过 HIV-PAH 患者的导管收集的 PA 内皮细胞中的 130/301-GLS1 轴。这些结果可以 在人类 HIV-PAH 中建立这一机制，并提出 HIV-PAH 检测所需的分子诊断方法。 目标 3) 确定 GLS1 的上调是否是促进 SIV-PAH 所必需的。在 SIV-PAH 猕猴中， 我们将施用 CB-839，这是一种正在人类癌症试验中测试的口服 GLS1 抑制剂，以确定其对 硬度、谷氨酰胺分解和 PAH。结果可以证明 GLS1 在 HIV 中的直接致病作用 相关的 PAH，因此可以重新利用该药物在人类 HIV 和 PAH 患者中进行快速、快速的试验。 意义：我们的团队在 HIV-PAH 的重大分子发现方面具有独特的优势。我们将 利用唯一已知可靠的 HIV-PAH 动物模型与人体研究，确保深入了解机制 以及对人类疾病的适用性达到了前所未有的程度。也许最重要的是，它提供了一种罕见的 为这种历史上被忽视的血管疾病建立急需的靶向治疗方法提供了机会。