Frataxin deficiency as a cause of endothelial senescence in multiple subtypes of pulmonary hypertension

Frataxin 缺乏是多种肺动脉高压亚型内皮衰老的原因

• 批准号: 10450703
• 负责人: Stephen Y Chan
• 金额: $ 61.99万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10450703
• 关键词: Ataxia; Automobile Driving; Binding; Binding Proteins; Biogenesis; Blood Vessels; Cardiomyopathies; Cell Aging; Cell Cycle Arrest; Cell Line; Cell Proliferation; Cells; Cellular biology; Confusion; DNA Damage; Data; Dependovirus; Development; Disease; Disease model; Dose; Endothelial Cells; Endothelium; Evolution; Friedreich Ataxia; Functional disorder; Funding; GSTP1 gene; Genetic; Genetically Engineered Mouse; Genomics; Genotoxic Stress; Heart Atrium; Heart Diseases; Hematological Disease; Heterogeneity; Histology; Human; Hypertrophic Cardiomyopathy; Hypoxia; Inflammation; Inflammatory; Interleukin-6; Knock-out; Knockout Mice; Left; Link; Lung; Metabolic dysfunction; Mitochondria; Modeling; Molecular; Mutation; Myelogenous; Myeloid Cells; Nuclear; Patients; Pharmaceutical Preparations; Phenotype; Plasma; Proteins; Pulmonary Hypertension; Pulmonary artery structure; Regulation; Reporting; Rodent; Role; Shapes; Shunt Device; Signal Transduction; Specimen; Stress; Sulfur; Sulofenur; Syndrome; Technology; Transgenic Organisms; Work; analog; cell type; cohort; cytokine; drug efficacy; endothelial stem cell; frataxin; genome editing; improved; in vivo; induced pluripotent stem cell; inhibitor; iron deficiency; mitochondrial metabolism; mouse model; novel; novel therapeutics; prevent; pulmonary arterial pressure; pulmonary vascular disorder; pulmonary vascular remodeling; replication stress; senescence; single-cell RNA sequencing; therapeutic development

Background: Endothelial cell (EC) pathobiology drives pulmonary hypertension (PH), but confusion over the evolution of EC phenotypes in this disease has persisted for decades. EC senescence, a state of stable cell cycle arrest, has been reported in PH, but the regulatory features are unknown. Led by our prior work showing deficiency of iron-sulfur (Fe-S) clusters in PH, we found that a Fe-S biogenesis protein, frataxin (FXN), controls senescence in pulmonary ECs. This may occur in Friedreich’s ataxia (FRDA), a disease marked by genetic FXN deficiency, cardiomyopathy, and often PH. Here, we offer a new model of EC biology in PH, where FXN loss promotes genotoxic stress and senescence in a pulmonary EC subset with low enough FXN. Senescent ECs then promote inflammation and drive many PH subtypes, including PH of FRDA and left heart disease. Hypothesis: We propose FXN deficiency, driven by genetic or acquired means, orchestrates Fe-S-dependent genotoxic stress, enforcing EC senescence and multiple PH subtypes. Aim 1. Determine if FXN deficiency drives DNA damage to enforce EC senescence. By study of human pulmonary artery and microvascular ECs and via genome editing of FXN mutations in inducible pluripotent stem cell (iPSC)-derived ECs from FRDA patients, we will study the role of FXN in genomic stress and EC senescence. Via study of circulating factors, histology, and single cell RNA sequencing, we will assess EC senescence in plasma and rare lung specimens from PAH patients and FRDA and HCM patients with pulmonary vascular disease. We expect to see a dose- dependent orchestration of FXN activities converging on EC senescence and PH. Aim 2. Determine if EC FXN deficiency depends upon senescence and myeloid inflammation to drive PH. In mice models of Groups 1 PAH and Group 2 PH due to FRDA, by using EC FXN-/- (KO) technology and adeno-associated virus delivery of FXN and its binding partner ISCU to pulmonary ECs in vivo, we will assess EC genomic stress, senescence, inflammation, and PH. EC-specific p16 KO mice and CX3CR1 KO mice will be used to define if EC FXN depends upon senescence and downstream myeloid inflammation to control PH. Thus, we aim to prove a new causative model of EC biology in PH – one that deconvolutes the confusion over EC heterogeneity that has plagued this field for decades. Aim 3. Determine if a novel GSTP1 inhibitor increases FXN and reverses multiple PH subtypes. We found that a GSTP1 inhibitor increases FXN and ISCU and improves PH. We will define this drug’s efficacy for ameliorating Groups 1-2 PH models and if FXN and GSTP1 are crucial for its function (via FXN and GSTP1/2 KO mice). If so, we could define an entirely new Fe-S-specific therapy for PH and FRDA. Significance: Via unique human and rodent discovery platforms, we will investigate an EC Fe-S cluster- senescence axis controlling multiple PH subtypes. Our work could explain the evolution of EC biology in PH and shift molecular paradigms, particularly for FRDA and Group 2 PH. Our work will also develop new therapies using senolytic drugs to control Fe-S biogenesis, notably applicable to Group 2 PH with no approved treatments.

背景：内皮细胞 (EC) 病理学驱动肺动脉高压 (PH)，但人们对内皮细胞 (EC) 病理学的混淆 这种疾病中 EC 表型的进化已经持续了几十年，这是一种稳定的细胞状态。 PH 中已报道了周期停滞，但根据我们之前的工作显示，其监管特征尚不清楚。 PH 中铁硫 (Fe-S) 簇的缺乏，我们发现 Fe-S 生物合成蛋白 frataxin (FXN) 可以控制 肺部内皮细胞的衰老可能发生在弗里德赖希共济失调（FRDA）中，这是一种以遗传为标志的疾病。 FXN 缺乏、心肌病以及常见的 PH 在这里，我们提供了 PH 中的 EC 生物学的新模型，其中 FXN。 FXN 足够低的情况下，损失会促进肺 EC 亚群的遗传毒性应激和衰老。 EC 随后会促进炎症并导致许多 PH 亚型，包括 FRDA 的 PH 和左心疾病。 假设：我们提出 FXN 缺乏，由遗传或后天途径驱动，协调 Fe-S 依赖性 基因毒性应激、强制 EC 衰老和多种 PH 亚型 目标 1. 确定 FXN 是否缺乏。 通过对人肺动脉和微血管 EC 的研究，驱动 DNA 损伤以增强 EC 衰老。 并通过对来自 FRDA 的诱导多能干细胞 (iPSC) 衍生的 EC 中的 FXN 突变进行基因组编辑 我们将通过循环因素研究 FXN 在基因组应激和 EC 衰老中的作用。 组织学和单细胞 RNA 测序，我们将评估血浆和稀有肺标本中的 EC 衰老 来自 PAH 患者以及患有肺血管疾病的 FRDA 和 HCM 患者，我们预计会看到剂量- FXN 活动的依赖协调与 EC 衰老和 PH 相关 目标 2. 确定 EC FXN 是否存在。 在第 1 组小鼠模型中，缺乏症取决于衰老和骨髓炎症来驱动 PH。 通过使用 EC FXN-/- (KO) 技术和腺相关病毒递送，由 FRDA 引起的 PAH 和 2 组 PH FXN 及其结合伙伴 ISCU 与体内肺 EC 的结合，我们将评估 EC 基因组应激、衰老、 炎症和 PH 特异性 p16 KO 小鼠和 CX3CR1 KO 小鼠将用于定义 EC FXN 是否依赖。 因此，我们的目标是证明一个新的病因。 PH 中的 EC 生物学模型 – 该模型消除了困扰这一问题的 EC 异质性的困惑 目标 3. 确定新型 GSTP1 抑制剂是否可以增加 FXN 并逆转多种 PH 我们发现 GSTP1 抑制剂可增加 FXN 和 ISCU 并改善 PH。 药物改善 1-2 组 PH 模型的功效以及 FXN 和 GSTP1 是否对其功能至关重要（通过 FXN 和 GSTP1/2 KO 小鼠）如果是这样，我们可以为 PH 和 FRDA 定义一种全新的 Fe-S 特异性疗法。 意义：通过独特的人类和啮齿动物发现平台，我们将研究 EC Fe-S 簇 - 我们的工作可以解释 PH 中 EC 生物学的进化。 并转变分子范式，特别是针对 FRDA 和第 2 组 PH，我们的工作还将开发新疗法。 使用 senolytic 药物控制 Fe-S 生物发生，特别适用于没有批准治疗的 2 组 PH。