Pulmonary Hypertension in Genetically Modified Mice

转基因小鼠的肺动脉高压

• 批准号: 7384373
• 负责人: Marlene Rabinovitch
• 金额: $ 50.18万
• 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 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7384373
• 关键词: Adult; Advanced Glycosylation End Products; Affect; Age; Apoptosis; Bleomycin; Blood Vessels; Calcium-Binding Proteins; Carrier Proteins; Cell physiology; Cells; Cellular Structures; Child; Clinical; DNA; Data; Dexfenfluramine; Disease; Elastases; Embryo; Functional disorder; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Gene Mutation; Genes; Genetic; Genetic Polymorphism; Genomics; Human; Hypoxia; Inflammation; Lead; Lesion; Life; Link; Lung; Lung diseases; Mediating; Microarray Analysis; Molecular; Molecular Abnormality; Molecular Profiling; Mouse Strains; Mus; Mutation; Neoplasm Metastasis; Pancreatic Elastase; Pathway interactions; Patients; Pattern; Predisposition; Process; Production; Protein Overexpression; Proteins; Pulmonary Circulation; Pulmonary Hypertension; Pulmonary artery structure; RNA Interference; RUNX1 gene; Recombinants; Relative (related person); Research Personnel; Resistance; Rodent Model; Serotonin; Severities; Signal Pathway; Signal Transduction; Signaling Pathway Gene; Smad Proteins; Smad protein; Smooth Muscle Myocytes; Stimulus; Structure; Tissues; Transgenic Mice; Vascular remodeling; Work; base; bone morphogenetic protein receptor type II; cell motility; genetic manipulation; insight; loss of function; lung injury; migration; postnatal; programs; pulmonary artery endothelial cell; receptor; response; serotonin transporter; tool; transcription factor; transdifferentiation; young adult

DESCRIPTION (provided by applicant): Idiopathic pulmonary hypertension (PH) is associated with progressive pulmonary vascular obliterative disease (PVOD) in otherwise healthy children and young adults. Some patients have an underlying genetic mutation causing loss of function of bone morphogenetic protein receptor II (BMP-RII) and some others have polymorphisms leading to heightened activity of the serotonin transporter (SERT). How these genetic abnormalities cause PVOD, is, however, unclear. Our recent studies in cultured human pulmonary artery (PA) smooth muscle cells (SMC) have shown that serotonin, likely in conjunction with SERT, increases expression of the calcium binding protein S100A4/Mts1 (Mts1). We further observed that recombinant Mts1 induces motility of PA SMC. Consistent with this, intense expression of Mts1 is apparent in PVOD in clinical tissue, in contrast to minimal expression in normal vessels. Also, 5-10% of transgenic mice that over-express Mts1 acquire PVOD as they age and all Mts1 overexpressing mice develop more severe PH than controls during hypoxia that does not regress with return to normoxia. Microarray analysis of DNA in lungs from Mts1 over-expressing vs. control mice shows a reduction in BMP-RI, the co-receptor of BMP-RII. We therefore propose that there are common and complementary molecular pathways linking BMP-RII, SERT and Mts1 in the pathophysiology of PVOD. Specific Aim I utilizes a variety of tools including short interference (si) RNA in cultured human PA endothelial and SMC to determine how BMP-RII, SERT and Mts1 might impact on vascular remodeling, either by inducing elastase activity and/or by promoting cell migration, proliferation, transdifferentiation, or propensity or resistance to apoptosis. These features will be correlated with the function of the transcription factor AML1, and with the genomic profiles of the cells. In Specific Aim II, transgenic mice with Mts1 over-expression will be investigated with respect to vascular reactivity and severity of PH in response to dexfenfluramine or inflammation. We also will produce and characterize a mouse with conditional global or SMC targeted deletion of BMP-RII, in terms of vascular cell structure, function, and propensity to PH. Genomic data from lung, endothelial and SMC from Mts1 over-expressing, BMP-RII deleted, and control mice will be correlated with function to identify common and complementary gene expression patterns related to PVOD. These studies should provide insights into the genetic basis for PVOD and information useful in developing strategies to arrest or reverse the process.

描述（由申请人提供）：特发性肺动脉高压（PH）与其他方面健康的儿童和年轻人的进行性肺血管闭塞性疾病（PVOD）相关。一些患者存在潜在的基因突变，导致骨形态发生蛋白受体 II (BMP-RII) 功能丧失，而另一些患者则存在多态性，导致血清素转运蛋白 (SERT) 活性增强。然而，这些基因异常如何导致 PVOD 尚不清楚。我们最近对培养的人肺动脉 (PA) 平滑肌细胞 (SMC) 的研究表明，血清素可能与 SERT 结合，增加钙结合蛋白 S100A4/Mts1 (Mts1) 的表达。我们进一步观察到重组 Mts1 诱导 PA SMC 的运动。与此一致的是，Mts1 在临床组织中的 PVOD 中明显强烈表达，而正常血管中的表达很少。此外，5-10% 过度表达 Mts1 的转基因小鼠随着年龄的增长而获得 PVOD，并且所有 Mts1 过度表达小鼠在缺氧期间会出现比对照更严重的 PH，并且不会随着恢复正常氧而消退。与对照小鼠相比，Mts1 过表达小鼠肺部 DNA 的微阵列分析显示 BMP-RI（BMP-RII 的共同受体）减少。因此，我们提出在 PVOD 的病理生理学中存在连接 BMP-RII、SERT 和 Mts1 的共同且互补的分子途径。具体目标 I 利用多种工具，包括培养的人 PA 内皮和 SMC 中的短干扰 (si) RNA，以确定 BMP-RII、SERT 和 Mts1 如何通过诱导弹性蛋白酶活性和/或促进细胞迁移来影响血管重塑、增殖、转分化、或对细胞凋亡的倾向或抵抗。这些特征将与转录因子 AML1 的功能以及细胞的基因组谱相关。在特定目标 II 中，将研究 Mts1 过度表达的转基因小鼠对右芬氟拉明或炎症反应的血管反应性和 PH 严重程度。我们还将在血管细胞结构、功能和 PH 倾向方面生产和表征具有条件性全局或 SMC 靶向删除 BMP-RII 的小鼠。来自 Mts1 过表达、BMP-RII 缺失和对照小鼠的肺、内皮和 SMC 的基因组数据将与功能相关联，以识别与 PVOD 相关的常见和互补基因表达模式。这些研究应该提供对 PVOD 遗传基础的见解，以及有助于制定阻止或逆转这一过程的策略的信息。