Pulmonary Hypertension in Genetically Modified Mice

转基因小鼠的肺动脉高压

• 批准号: 10337210
• 负责人: Marlene Rabinovitch
• 金额: $ 75.31万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10337210
• 关键词: 3-Dimensional; ALDH1A2 gene; Abnormal Cell; Acetyl Coenzyme A; Address; Affect; Agonist; Anti-Inflammatory Agents; Apoptosis; Arachidonate 5-Lipoxygenase; Attenuated; BMPR2 gene; Blood Vessels; Breeding; Cell Communication; Cell Cycle Progression; Cell Proliferation; Cell physiology; Cells; Cellular Metabolic Process; ChIP-seq; Chromatin; Chronic; Clinic; Confocal Microscopy; Cultured Cells; Data; Disease; Endothelial Cells; Endothelium; Enhancers; Enzymes; Epigenetic Process; Exposure to; Family; Funding; Gene Abnormality; Gene Expression; Gene Expression Regulation; Genetic Transcription; Grant; Histone Acetylation; Histones; Human; Hypoxia; Image; Inflammation; Inflammatory; Inflammatory Response; Inherited; Injury; Lead; Ligands; Link; LoxP-flanked allele; Lung; Mediating; Mesenchymal; Messenger RNA; Metabolic; Metabolic Pathway; Metabolism; Mitochondria; Mus; Muscle; Mutation; Nature; Nuclear; Oxidants; Pathogenesis; Patients; Penetrance; Production; Protein Isoforms; Proteins; Proteomics; Pulmonary Hypertension; Pulmonary arterial remodeling; Regulation; Reporting; Resolution; Role; Severities; Signal Transduction; Smooth Muscle Myocytes; Structure of parenchyma of lung; Testing; Time; Tissues; Transferase; aldehyde dehydrogenases; antagonist; bone morphogenetic protein receptors; cancer cell differentiation; cdc Genes; chromatin remodeling; experimental study; genetically modified cells; hypoxia-induced pulmonary hypertension; improved; improved functioning; interstitial; macrophage; member; metabolomics; mouse model; novel; novel therapeutics; oxidant stress; preservation; primary pulmonary hypertension; pulmonary arterial hypertension; pulmonary artery endothelial cell; receptor function; recruit; response; response to injury; small molecule; transcriptome sequencing; treatment response

This renewal application builds upon our recent studies indicating that aldehyde dehydrogenase (ALDH) enzymes can be critical metabolic switches that link chromatin remodeling with gene expression in vascular and inflammatory cells, and that the dysregulation of specific ALDH isoforms is linked in the pathogenesis of pulmonary arterial hypertension (PAH). We use pulmonary arterial (PA) endothelial cells (EC), smooth muscle cells (SMC) and macrophages (MØ) and genetically modified mice to pursue our novel observations. Our Preliminary Data reveal increased mRNA and protein levels of ALDH1A3 in PA SMC from patients with PAH versus controls, which is required for their heightened proliferation. The mechanism is related to elevated nuclear production of acetyl CoA, acetylation of histones at enhancer marks (H3K27ac) and expression of genes associated with cell cycle progression. In Aim 1, we use RNA Seq and ChIP Seq to probe the mechanism by which elevated ALDH1A3 increases cell cycle genes as well as other metabolic enzymes, i.e., PKM2, DLD and IDH1 and we study the impact of those enzymes on chromatin remodeling and gene expression. We investigate whether elevated ALDH1A3 could be regulated by mechanisms that are dependent a well as independent of BMPR2. Further experiments test whether deleting Aldh1a3 in SMC of genetically modified mice is sufficient to attenuate proliferation of PA SMC and pulmonary hypertension (PH) associated with chronic hypoxia. In PA EC, we found that a different ALDH isoform ALDH3A1, is increased in response to laminar flow. In contrast, ALDH2 a mitochondrial enzyme is elevated under static or disturbed flow conditions. Both ALDH isoforms are expected to preserve PA EC function through multiple metabolic pathways that affect gene expression. Aim 2 will determine whether ALDH3A1 protects PA EC under laminar flow and ALDH2 under static conditions by relating their metabolomic profile to chromatin accessibility and gene expression. Localization of these ALDH isoforms will be studied in control and PAH lungs using 3-D vibratome imaging. Aldh3a1 or Aldh2 will be deleted in EC from genetically modified mice to determine whether this causes more severe PH following chronic hypoxia or 5-lipoxygenase mediated inflammation. In Aim 3 we focus on ALDH1A2, an ALDH isoform implicated in polarization of MØ associated with resolution of inflammation, similar to the reported effect of BMPR2 ligands. We will determine whether deleting Aldh1a2 or Bmpr2 in interstitial MØ recruited to the lung in association with PH producing conditions, causes more severe disease related to persistent perivascular inflammation. Proteomics will be applied to analyze the secretome of these Aldh1a2 or Bmpr2 deleted MØ to better define the nature of the inflammatory response. The impact of the MØ with Aldh1a2 or Bmpr2 deleted on EC apoptosis or endothelial mesenchymal transition will also be investigated. Our studies are timely in addressing the significance of ADLH isoforms in PAH as small molecule agonists and antagonists of ALDH enzymes are being developed and tested in the clinic.

这种更新应用是在我们最近的研究基础上建立的，表明醛脱氢酶（ALDH） 酶可以是关键的代谢开关，将染色质重塑与血管中的基因表达联系起来 和炎性细胞，并且特定ALDH同工型的失调与 肺动脉高压（PAH）。我们使用肺动脉（PA）内皮细胞（EC），平滑肌 细胞（SMC）和巨噬细胞（Mø）和转基因小鼠，以追求我们的新观测。我们的 初步数据显示，PA SMC中PA SMC患者的mRNA和蛋白质水平升高 与对照相比，这是其增殖加剧所必需的。该机制与升高有关 乙酰COA的核产生，在增强子标记处组蛋白的乙酰化（H3K27AC）和表达 与细胞周期进程相关的基因。在AIM 1中，我们使用RNA SEQ和CHIP SEQ探测 升高ALDH1A3的机制增加了细胞周期基因以及其他代谢酶，即 PKM2，DLD和IDH1，我们研究了这些酶对染色质重塑和基因的影响 表达。我们研究是否可以通过依赖机制来调节升高的AldH1A3 与BMPR2无关。进一步的实验测试是否在SMC中删除ALDH1A3是否一般 改良的小鼠足以减弱PA SMC的增殖和相关的肺动脉高压（pH） 患有慢性缺氧。在PA EC中，我们发现不同的ALDH同工型ALDH3A1因响应于 层流。相反，在静态或干扰的流动条件下，AldH2 A线粒体酶升高。 两种ALDH同工型都可以通过影响多种代谢途径来保留PA EC的功能 基因表达。 AIM 2将确定ALDH3A1是否在层流下保护PA EC和ALDH2 在静态条件下，通过将其代谢组谱与染色质访问性和基因表达联系起来。 这些ALDH同工型的定位将在对照中进行研究，并使用3-D振动型成像进行对照和PAH肺。 ALDH3A1或ALDH2将从转基因小鼠中删除EC中，以确定这是否导致更多 慢性缺氧或5-脂氧合酶介导的注射后严重的pH。在目标3中，我们专注于 ALDH1A2，一种在与感染分辨率相关的Mø的极化中实现的ALDH同工型 据报道BMPR2配体的作用。我们将确定在间隙中删除ALDH1A2或BMPR2是否 与pH产生条件相关的Mø招募到肺部，引起与 持续性周围感染。蛋白质组学将用于分析这些aldh1a2或 BMPR2删除了Mø，以更好地定义炎症反应的性质。 Mø与 还将研究在EC凋亡或内皮间质转变上删除的ALDH1A2或BMPR2。 我们的研究及时解决了ADLH同工型在PAH中的重要性，作为小分子激动剂和 ALDH酶的拮抗剂正在诊所开发和测试。

项目成果

Loss of adenomatous poliposis coli-α3 integrin interaction promotes endothelial apoptosis in mice and humans.

• DOI: 10.1161/circresaha.112.267849
• 发表时间: 2012-12-07
• 期刊: Circulation research
• 影响因子: 20.1
• 作者: de Jesus Perez VA;Yuan K;Orcholski ME;Sawada H;Zhao M;Li CG;Tojais NF;Nickel N;Rajagopalan V;Spiekerkoetter E;Wang L;Dutta R;Bernstein D;Rabinovitch M
• 通讯作者: Rabinovitch M

Right heart imaging in patients with heart failure: a tale of two ventricles.

• DOI: 10.1097/hco.0000000000000315
• 发表时间: 2016-09
• 期刊: Current opinion in cardiology
• 影响因子: 2.3
• 作者: Amsallem M;Kuznetsova T;Hanneman K;Denault A;Haddad F
• 通讯作者: Haddad F

Development of pulmonary arterial hypertension in mice over-expressing S100A4/Mts1 is specific to females.

• DOI: 10.1186/1465-9921-12-159
• 发表时间: 2011-12-20
• 期刊: Respiratory research
• 影响因子: 5.8
• 作者: Dempsie Y;Nilsen M;White K;Mair KM;Loughlin L;Ambartsumian N;Rabinovitch M;Maclean MR
• 通讯作者: Maclean MR