Novel roles of RNA modifications in the pathogenesis of pulmonary vascular remodeling and PAH

RNA修饰在肺血管重塑和PAH发病机制中的新作用

基本信息

批准号：
10540134
负责人：
YOU-YANG ZHAO
金额：
$ 68万
依托单位：
LURIE CHILDREN'S HOSPITAL OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10540134
关键词：
Address Adenosine Adult Affect Animal Model Area Arginine CRISPR/Cas technology CXCL12 gene Cessation of life Chronic Clinical Clustered Regularly Interspaced Short Palindromic Repeats Coupled Data Dioxygenases Disease Endothelial Cells Endothelium Epigenetic Process FDA approved Fatty acid glycerol esters Functional disorder Gene Expression Gene Transfer Genes Goals Human Hypoxia Impairment Interleukin-6 Knockout Mice Link Lung Malignant Neoplasms Meclofenamic Acid Mediating Messenger RNA Metabolism Modeling Modification Molecular Monocrotaline Morbidity - disease rate Mus Nitric Oxide Obesity PDGFB gene Pathogenesis Patients Pharmaceutical Preparations Pharmacology Phenotype Platelet-Derived Growth Factor Post-Transcriptional Regulation Prevention Proteins Protocols documentation Pulmonary Vascular Resistance RNA RNA Stability RNA metabolism Rattus Regulation Regulator Genes Research Role S-Adenosylhomocysteine Site Stimulus Structure of parenchyma of lung Therapeutic Therapeutic Agents Therapeutic Effect Time Transcription Coactivator Translations Vascular Endothelial Cell Vascular remodeling alpha ketoglutarate arginase base bioprocess clinically relevant demethylation druggable target effective therapy endothelial dysfunction human disease in vivo inhibitor insight mRNA Stability methyl group mortality mutant nanoparticle new therapeutic target next generation sequencing novel novel therapeutic intervention novel therapeutics premature primary pulmonary hypertension pulmonary arterial hypertension pulmonary vascular remodeling pulmonary vasoconstriction resistance gene response right ventricular failure synergism therapeutic evaluation transcription factor translational potential vasoconstriction

项目摘要

Pulmonary arterial hypertension (PAH) is characterized by progressive increase of pulmonary vascular resistance and obliterative vascular remodeling that causes right heart failure and premature death. Given the underlying molecular mechanisms of obliterative vascular remodeling remain enigmatic, current therapies have not targeted the fundamental disease modifying mechanisms and hence only resulted in a modest improvement in the morbidity and mortality. While several transcription factors and transcriptional co- activators have been studied in the context of PAH, post-transcriptional regulations of mRNAs that can affect expression of key proteins remain largely unexplored. RNA modifications including N6-methyladenosine (m6A) have recently been discovered as essential regulators of gene expression. The m6A modification controls RNA stability, transport, and translation and has been linked to human diseases such as obesity and cancers. Despite its functional importance in various fundamental bioprocesses, studies of m6A modification of mRNAs in PAH are lacking. Our Supporting Data show that expression of fat mass and obesity-associated protein (FTO), a well-characterized RNA demethylase, is markedly elevated in pulmonary vascular endothelial cells (ECs) of idiopathic PAH patients. Tie2Cre-mediated deletion of Fto in ECs (FtoTie2Cre) inhibited PH induced by chronic hypoxia. Our mechanistic studies provide evidence that several PAH-causing genes are potential FTO targets in human lung ECs. Furthermore, pharmacological inhibition of FTO in monocrotaline- challenged rats inhibited pulmonary vascular remodeling and PH. Thus, we hypothesize that endothelial FTO, as a major m6A eraser, acts as a key regulator of mRNA stability and accumulation of key PAH-causing genes in ECs to regulate pulmonary vasoconstriction and vascular remodeling and thus is a novel therapeutic target for PAH. We propose the following 3 Specific Aims. In Aim 1, we will define the fundamental role of endothelial FTO in the pathogenesis of PAH. In Aim 2, we will delineate the molecular mechanisms underlying FTO regulation of endothelial dysfunction leading to pulmonary vasoconstriction and vascular remodeling. We will identify the key FTO targets in ECs and address the possibility of rescuing the phenotype of FtoTieCre by novel nanoparticle-mediated in vivo EC-specific gene transfer. In Aim 3, we will explore the therapeutic potential of FTO inhibitors including a repurposed FDA-approved drug in the treatment of PAH employing 3 complimentary animal models of PAH. Based on the clear clinical relevance of our novel findings, we expect that the proposed studies have significant translational potential by delineating the molecular and cellular mechanisms of endothelial dysfunction leading to pulmonary vasoconstriction and vascular remodeling, identifying druggable targets, and exploring pharmacological agents that can pharmacologically inhibit/reverse vascular remodeling and also inhibit vasoconstriction for the prevention and treatment of PAH in patients. Thus, we believe the proposed studies have great translational potential.

肺动脉高压（PAH）的特点是肺血管进行性增加阻力和闭塞性血管重塑导致右心衰竭和过早死亡。给定闭塞性血管重塑的潜在分子机制仍然是个谜，目前的治疗方法没有针对基本的疾病改变机制，因此只产生了适度的效果发病率和死亡率的改善。虽然一些转录因子和转录共激活剂已在 PAH 背景下进行了研究，mRNA 的转录后调控可影响关键蛋白的表达在很大程度上仍未被探索。 RNA 修饰包括 N6-甲基腺苷 (m6A) 最近被发现是基因表达的重要调节因子。 m6A改装控制 RNA 稳定性、运输和翻译，并与肥胖和肥胖等人类疾病有关。癌症。尽管 m6A 修饰在各种基本生物过程中具有重要的功能，但其研究 PAH 中缺乏 mRNA。我们的支持数据表明，脂肪量和肥胖相关的表达蛋白（FTO）是一种充分表征的 RNA 去甲基化酶，在肺血管内皮细胞中显着升高特发性 PAH 患者的细胞（EC）。 Tie2Cre 介导的 EC 中 Fto 缺失 (FtoTie2Cre) 抑制 PH 慢性缺氧所致。我们的机制研究提供的证据表明，多种 PAH 致病基因是人肺 EC 中潜在的 FTO 靶点。此外，野百合碱中 FTO 的药理学抑制作用受攻击的大鼠抑制肺血管重塑和 PH。因此，我们假设内皮细胞 FTO 作为主要的 m6A 擦除器，充当 mRNA 稳定性和引起 PAH 的关键物质积累的关键调节因子 EC 中的基因可调节肺血管收缩和血管重塑，因此是一种新的治疗方法 PAH 目标。我们提出以下 3 个具体目标。在目标 1 中，我们将定义以下基本作用：内皮 FTO 在 PAH 发病机制中的作用在目标 2 中，我们将描述潜在的分子机制 FTO 调节内皮功能障碍，导致肺血管收缩和血管重塑。我们将确定 EC 中的关键 FTO 靶点并解决挽救 FtoTieCre 表型的可能性通过新型纳米颗粒介导的体内 EC 特异性基因转移。在目标 3 中，我们将探索治疗方法 FTO 抑制剂的潜力，包括 FDA 批准的药物的重新利用，用于治疗 PAH，采用 3 PAH 的补充动物模型。基于我们的新发现的明确临床相关性，我们预计所提出的研究通过描绘分子和细胞具有显着的转化潜力内皮功能障碍导致肺血管收缩和血管重塑的机制，识别可成药的靶点，并探索可药理学的药物制剂抑制/逆转血管重塑并抑制血管收缩以预防和治疗 PAH 在患者中。因此，我们相信所提出的研究具有巨大的转化潜力。