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）是一种依赖多种血管细胞的致命疾病 但是，分子串扰的关键系统仍然是个谜。在之前的奖项中，我们定义了一个关键的监管系统。 转录共激活子 YAP/TAZ 与谷氨酰胺酶 (GLS1) 之间的轴，建立了新的 然而，关键问题仍然存在。 激活 YAP/TAZ 引发 PAH 的触发因素是什么？它们是否源自不同的细胞类型？ 在这些触发因素的下游，其他氨基酸的代谢是否控制血管硬化和 PAH？ 最近，内皮细胞（EC）衰老-稳定的细胞周期停滞导致炎症信号通过 衰老相关分泌表型 (SASP) 因素在 PAH 中已有报道，但其后果 PAH 中的衰老机制尚未被探索，我们推测 EC 衰老会诱导炎症 SASP 信号传导。 PA 成纤维细胞，重新编程丝氨酸和谷氨酰胺代谢，以控制胶原沉积、血管 目标 1) 定义 EC 衰老在控制成纤维细胞谷氨酰胺和丝氨酸中的作用。 我们计划研究携带 EC 特异性缺陷的 PAH 小鼠。 衰老驱动程序 p16 以及对成纤维细胞 YAP 和下游代谢重编程的影响。 特定的分泌组追踪患有 PAH 的小鼠，我们将定义源自 SASP 蛋白因子的完整谱 PAH 相关的衰老 ECs。通过标记谷氨酰胺/丝氨酸后对人 PAH 肺进行单细胞 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 目标 2) 确定 GLS1 和丝氨酸分解代谢酶 SHMT1 是否发生改变。 在这里，我们将确定是否成纤维细胞特异性敲除 GLS1。 或 SHMT1 逆转 PAH 小鼠的血管硬化，并且如果 AAV 特异性递送 SHMT1 和 GLS1 驱动 使用小分子抑制 PLGA 中封装的 YAP/GLS1/SHMT1。 纳米粒子用于吸入疗法，我们将确定这种疗法逆转血管硬化和 目标 3) 利用 18F-氟谷氨酰胺 PET 成像来测量 SSc-PAH 与 SSc-PAH 中的谷氨酰胺摄取量。 我们将在系统性硬化症依赖性 PAH (SSc-PAH) 和 SSc 中测试 18F-FGln PET 成像。 患有早期 PAH 的患者，运动 PH 本研究将确定谷氨酰胺的相关性。 人类 PAH 发展（不仅仅是终末阶段）的代谢以及 18F-FGln 发挥作用的潜力 一种新型 SSc-PAH 诊断示踪剂 意义：我们的多学科团队在定义方面具有独特的优势。 EC 衰老到成纤维细胞的代谢途径对于诱导血管硬化和 PAH 至关重要。 测试一种新型吸入组合代谢疗法，我们将开展首次人体诊断研究 18F-FGln PET/CT 因此，我们的目标是建立汇聚于成纤维细胞氨基的广泛细胞间轴。 酸代谢作为 PAH 的关键调节剂，从而提供新颖的靶向治疗和诊断。