An endothelial-fibroblast axis connecting senescence to amino acid metabolism for control of vascular stiffness in PAH

连接衰老与氨基酸代谢以控制 PAH 血管僵硬度的内皮-成纤维细胞轴

• 批准号: 10378309
• 负责人: Stephen Y Chan
• 金额: $ 79万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10378309
• 关键词: 18F-Glutamine; Amino Acids; Automobile Driving; Award; Blood Vessels; Catabolism; Cell Aging; Cell Cycle Arrest; Cells; Collagen; Critical Pathways; Deposition; Detection; Development; Diagnostic; Disease; Encapsulated; Endothelial Cells; Endothelium; Enzymes; Etiology; Event; Exercise; Feedback; Fibroblasts; Funding; Genetic; Glutaminase; Glutamine; Human; Imaging Techniques; Inflammatory; Ingestion; Inhalation; Inhalation Therapy; Isotopes; Knock-out; Knockout Mice; Label; Link; Lung; Maintenance; Measures; Mediating; Metabolic; Metabolism; Modeling; Molecular; Mus; PET/CT scan; Pathogenicity; Patients; Pharmacotherapy; Phenotype; Positioning Attribute; Positron-Emission Tomography; Proteins; Pulmonary vessels; Reporting; Right ventricular structure; Rodent; Role; Serine; Shapes; Signal Transduction; Sum; System; Systemic Scleroderma; Testing; Therapeutic; Tissues; Tracer; Transcription Coactivator; Vascular Proliferation; Work; amino acid metabolism; cell type; combinatorial; drug inhalation; first-in-human; human RNA sequencing; human study; mass spectrometric imaging; multidisciplinary; nanoparticle; new therapeutic target; novel; novel diagnostics; pulmonary arterial hypertension; senescence; single-cell RNA sequencing; small molecule; spectrograph; targeted treatment; therapeutically effective; uptake

Background: Pulmonary arterial hypertension (PAH) is a deadly disease dependent on several vascular cell types. But, key systems of molecular cross-talk remain enigmatic. In the prior award, we defined a key regulatory axis between the transcriptional coactivators YAP/TAZ with the enzyme glutaminase (GLS1), establishing a new paradigm of how glutamine metabolism is related to vascular stiffness in PAH. Yet, crucial questions remain. What are the triggers that activate YAP/TAZ to initiate PAH and do they originate from separate cell types? Downstream of those triggers, does metabolism of other amino acids control vascular stiffening and PAH? Recently, endothelial cell (EC) senescence–stable cell cycle arrest resulting in inflammatory signaling via senescence associated secretory phenotype (SASP) factors–was reported in PAH, but the consequences of senescence in PAH are unexplored. We postulate that EC senescence induces inflammatory SASP signaling to PA fibroblasts, reprogramming serine along with glutamine metabolism to control collagen deposition, vascular stiffness, and PAH. Aim 1) Define the role of EC senescence in controlling fibroblast glutamine and serine metabolism, vascular stiffening, and PAH. We plan to study PAH mice carrying EC-specific deficiency of the senescence driver p16 and the effects on fibroblast YAP and downstream metabolic reprogramming. Via EC- specific secretome-tracking mice with PAH, we will define the entire profile of SASP protein factors derived from PAH-relevant senescent ECs. By single cell RNA sequencing of human PAH lung after labeled glutamine/serine ingestion and spectral (MIMS) imaging, we will determine if EC senescence correlates with fibroblast glutamine/serine uptake. Aim 2) Determine if alterations of GLS1 and the serine catabolism enzyme SHMT1 are essential for vascular stiffening and PAH. Here, we will determine if fibroblast-specific knockout of GLS1 or SHMT1 reverses vascular stiffening in PAH mice and if AAV-specific delivery of SHMT1 and GLS1 drives vascular stiffening and PAH. Using small molecules to inhibit YAP/GLS1/SHMT1 encapsulated in PLGA nanoparticles for inhaled therapy, we will define the efficacy of such therapy to reverse vascular stiffening and PAH. Aim 3) Utilize 18F-fluoroglutamine PET imaging to measure glutamine uptake in SSc-PAH vs. controls. We will test 18F-FGln PET imaging in systemic sclerosis-dependent PAH (SSc-PAH) and in SSc patients with an early-stage form of the PAH, exercise PH. This study will define the relevance of glutamine metabolism in the development (not merely end-stage) of human PAH and the potential of 18F-FGln to serve as a novel diagnostic tracer for SSc-PAH. Significance: Our multi-disciplinary team is uniquely positioned to define an EC senescence-to-fibroblast metabolism pathway critical for inducing vascular stiffening and PAH. We will test a novel inhaled combinatorial metabolic therapy, and we will embark on a first-in-human diagnostic study of 18F-FGln PET/CT. Thus, we aim to establish the broad intercellular axes that converge upon fibroblast amino acid metabolism as a crucial regulator of PAH, thereby offering novel targeted therapeutics and diagnostics.

背景：肺动脉高压（PAH）是一种致命疾病，取决于几个血管细胞 类型。但是，分子串扰的关键系统仍然神秘。在先前的奖项中，我们定义了一个关键的监管 转录共激活剂yap/taz与酶谷氨酰胺酶（GLS1）之间的轴，建立了新的 谷氨酰胺代谢与PAH中的血管刚度如何相关的范例。但是，仍然存在关键问题。 激活YAP/TAZ以启动PAH的触发器是什么，它们源自单独的细胞类型？ 这些触发因素的下游，其他氨基酸的代谢是否控制着血管僵硬和PAH？ 最近，内皮细胞（EC）感应 - 稳定的细胞周期停滞，导致炎症信号传导通过 相关的相关秘密表型（SASP）因子 - 在PAH中报道了，但是 PAH中的衰老是出乎意料的。我们假设EC衰老诱导炎症SASP信号传导 PA成纤维细胞，重编程系列以及谷氨酰胺代谢以控制胶原蛋白沉积，血管 僵硬和pah。目标1）定义EC感应在控制成纤维细胞谷氨酰胺和系列中的作用 代谢，血管僵硬和PAH。我们计划研究带有EC特异性缺陷的PAH小鼠 感应驱动器P16以及对成纤维细胞YAP和下游代谢重编程的影响。通过ec- 使用PAH进行特定的分泌组跟踪小鼠，我们将定义SASP蛋白质因子的整个特征 与PAH相关的感觉ECS。通过标记为谷氨酰胺/丝氨酸的人PAH肺的单细胞RNA测序 摄入和光谱（MIMS）成像，我们将确定EC衰老是否与成纤维细胞相关 谷氨酰胺/丝氨酸摄取。目标2）确定GLS1和丝氨酸分解代谢酶SHMT1的改变是否 对于血管僵硬和PAH是必不可少的。在这里，我们将确定GLS1的成纤维细胞特异性敲除 或SHMT1逆转PAH小鼠的血管僵硬，以及SHMT1和GLS1驱动的AAV特定递送 血管僵硬和PAH。使用小分子抑制封装在PLGA中的YAP/GLS1/SHMT1 纳米颗粒用于遗传治疗，我们将定义这种疗法逆转血管僵硬和 pah。目标3）利用18F-氟谷氨酰胺PET成像来测量SSC-PAH VS中的谷氨酰胺摄取。 控件。我们将在全身性硬化症依赖性PAH（SSC-PAH）和SSC中测试18F-FGLN PET成像 PAH的早期形式的患者，运动pH。这项研究将定义谷氨酰胺的相关性 人类PAH的发展（不仅是终阶段）的代谢和18f-FGLN的潜力 SSC-PAH的新型诊断示踪剂。意义：我们的多学科团队在定义的独特位置 EC衰老到纤维细胞代谢途径对于诱导血管僵硬和PAH至关重要。我们将 测试一种新型的遗传组合代谢疗法，我们将开始对 18F-FGLN PET/CT。这是，我们的目的是建立在成纤维细胞氨基上收敛的宽细胞间轴 酸代谢是PAH的关键调节剂，从而提供了新颖的靶向疗法和诊断。