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

背景：肺动脉高压(PAH)是一种发病率增加的致命性血管疾病。 感染人类免疫缺陷病毒(HIV)。艾滋病毒-多环芳烃可能比最初更普遍 对于这种神秘的PAH形式，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在PAH中是相互关联的过程，包括灵长类和人类 艾滋病毒-多环芳烃的例子。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，miR-130/301、YAP或 GLS1改善PAH。我们还发现，感染HIV的T细胞释放无细胞的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，从而激活血管僵硬、谷氨酰胺分解和HIV-PAH。 具体目的：目的1)确定HIV感染的T细胞释放的miR-21是否诱导肺血管 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 HIV感染的T细胞对肺血管细胞的MIR-21。在体内，使用miR-21-Fulete的输注 与miR-21耗尽的血浆进入miR-21-/-小鼠与野生型小鼠，我们将确定循环中的miR-21是否 传递到肺内皮细胞，并诱导血管僵硬，谷氨酰胺分解，和PAH。结果可能是 在HIV感染和肺血管谷氨酰胺分解和僵硬之间建立一个全新的miR联系。 目的2)确定HIV-PAH患者是否存在明显的肺动脉僵硬和谷氨酰胺分解反应。 从先前收集的HIV-PAH样本中，我们将把血浆miR-21水平与肺动脉(PA)联系起来。 根据血液动力学数据和反映PA谷氨酰胺分解反应的血浆代谢物计算顺应性。vbl.使用 优化技术基于我们公布的协议，我们还将评估YAP/TAZ-miR的激活- 经导管采集的HIV-PAH患者PA内皮细胞130/301-GLS1轴。这些结果可能 在人类HIV-PAH中建立这一机制，并建议对HIV-PAH检测进行必要的分子诊断。 目的3)确定GLS1上调是否是促进SIV-PAH所必需的。在SIV-PAH猕猴中， 我们将服用CB-839，一种正在进行人类癌症试验的口服GLS1抑制剂，以确定其对 僵硬、谷氨酰胺分解和PAH。结果表明，GLS1在HIV中具有直接致病作用。 因此，这种药物可以重新用于在人类艾滋病毒和多环芳烃患者身上进行快速、快速的试验。 意义：我们的团队在HIV-PAH的重大分子发现方面处于独特的地位。我们会 利用已知唯一可靠的艾滋病毒-多环芳烃动物模型进行人体研究，确保机械洞察力 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 为这一历史上被忽视的血管疾病建立亟需的靶向治疗的机会。

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.