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

背景：肺动脉高压（PAH）是一种发病率不断上升的致命性血管疾病 人类免疫缺陷病毒（HIV）感染。HIV-PAH可能比原来更加流行 然而，对于这种神秘形式的PAH，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的相关过程，包括灵长类动物和人类 HIV-PAH的例子。转录共激活因子雅普/TAZ诱导microRNA（miR）-130/301家族 和谷氨酰胺酶（GLS 1）来控制这些表型。在PAH啮齿动物模型中，miR-130/301、雅普或 GLS 1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 共培养肺血管细胞中的组胺分解和基质重塑途径。我们假设 雅普/TAZ-miR-130/301-GLS 1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、组胺溶解和HIV-PAH。 1）确定从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，我们将研究的直接交付和行动， miR-21对来自HIV感染的T细胞的肺血管细胞的作用。在体内，采用输注充满miR-21的 vs.将miR-21-/-小鼠与野生型小鼠的血浆中的miR-21去除后，我们将确定循环miR-21是否 递送至肺内皮并诱导血管硬化、组胺溶解和PAH。结果可能 在HIV感染与肺血管平滑肌溶解和僵硬之间建立全新的miR联系。 目的2）确定肺动脉僵硬和氨氯地平溶解是否在患有HIV-PAH的人中明显。 从先前收集的HIV-PAH样本中，我们将血浆miR-21水平与肺动脉（PA） 根据血流动力学数据和血浆代谢产物计算的顺应性，反映PA β-内酰胺酶解。使用 基于我们公布的方案优化的技术，我们还将评估雅普/TAZ-miR-1的激活。 130/301-GLS 1轴在PA内皮细胞通过导管收集的HIV-PAH患者。这些结果可能 在人类HIV-PAH中建立这种机制，并提出HIV-PAH检测所需的分子诊断。 目的3）确定GLS 1的上调是否是促进SIV-PAH所必需的。在SIV-PAH猕猴中， 我们将给予CB-839，一种正在人类癌症试验中测试的口服GLS 1抑制剂，以确定其对 僵硬、肌氨溶解和PAH。结果可以证明GLS 1在HIV-1中的直接致病作用。 因此，可以将这种药物重新用于人类HIV和PAH患者的快速加速试验。 意义：我们的团队在发现HIV-PAH的主要分子方面具有独特的优势。我们将 利用人类研究中唯一已知的可靠的HIV-PAH动物模型，确保了解机制 和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.