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

基本信息

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
项目类别：
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

项目摘要

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），但混乱这种疾病中EC表型的演变持续了数十年。 EC感应，稳定细胞的状态 pH中有循环停滞，但调节特征尚不清楚。由我们先前的工作领导铁硫（Fe-S）簇的缺乏，我们发现Fe-S生物发生蛋白Frataxin（FXN）对照肺EC中的感应。这可能发生在弗里德里希共济失调（FRDA）中，这种疾病以遗传为标志 FXN缺乏症，心肌病和通常pH。在这里，我们在pH中提供了一种新的EC生物学模型，其中FXN 损失促进了足够低的FXN的肺EC子集中的遗传毒性应力和感应。衰老然后，EC促进注射并驱动许多pH亚型，包括FRDA和左心脏病。假设：我们提出了FXN缺乏症，由遗传或获得的手段驱动，协调Fe-S依赖性遗传毒性应激，实施EC衰老和多个pH亚型。目标1。确定FXN是否缺乏驱动DNA损伤以实施EC感应。通过研究人类肺动脉和微血管ECS 并通过FXN突变的基因组编辑诱导多能干细胞（IPSC）衍生的EC中的FXN突变。患者，我们将研究FXN在基因组应激和EC衰老中的作用。通过研究循环因素，组织学和单细胞RNA测序，我们将评估血浆和罕见肺标本中的EC感应来自PAH患者，FRDA和HCM患有肺血管疾病的患者。我们希望看到剂量 - FXN活性的依赖管弦在EC感应和pH上融合。目标2。确定EC FXN是否缺乏取决于感应和髓样注射以驱动pH。在组1的小鼠模型中 PAH和第2组pH是由于FRDA，使用EC FXN - / - （KO）技术和腺相关病毒输送 FXN及其结合伴侣ISCU与体内肺EC的ISCU，我们将评估EC基因组应力，衰老，炎症和pH。 EC特异性P16 KO小鼠和CX3CR1 KO小鼠将用于定义EC FXN是否依赖感应和下游髓样注射后，以控制pH。这，我们旨在证明一种新的因果关系 pH中的EC生物学模型 - 一种涉及困扰EC异质性的混淆的模型数十年来。目标3。确定新型的GSTP1抑制剂是否增加了FXN并逆转多个pH 亚型。我们发现GSTP1抑制剂增加了FXN和ISCU并改善pH值。我们将定义这个药物对改善1-2个pH模型的效率以及FXN和GSTP1的功能至关重要（VIA） FXN和GSTP1/2 KO小鼠）。如果是这样，我们可以为PH和FRDA定义全新的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，我们将研究一个EC FE-S群集 - 控制多个pH亚型的感应轴。我们的工作可以解释pH中EC生物学的演变并移动分子范式，特别是对于FRDA和第2组pH。我们的工作还将开发新的疗法使用鼻溶剂来控制Fe-S生物发生，特别是适用于未经批准治疗的2组pH。