Morphogenesis of the pulmonary artery smooth muscle layer

肺动脉平滑肌层的形态发生

• 批准号: 7511793
• 负责人: Daniel Greif
• 金额: $ 13.21万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7511793
• 关键词: 5-bromo-4-chloro-3-indolyl beta-galactoside; Angioplasty; Apoptosis; Arteries; Atherosclerosis; Biological Assay; Biological Markers; Blood Vessels; Boxing; Cell Differentiation process; Cell Proliferation; Cell physiology; Cells; Confocal Microscopy; Coronary artery; Development; Development, Other; Disease; Elements; Embryo; Endothelial Cells; Event; Functional disorder; Goals; Hypertension; In Situ Hybridization; In Vitro; Indium; Investigation; Knockout Mice; LacZ Genes; Left; Left pulmonary artery; Ligands; Light; Lung; Lymphangioleiomyomatosis; Maintenance; Maps; Mediating; Mesenchymal; Molecular; Morphogenesis; Mus; Nutrient; Other Finding; Pathogenesis; Pathology; Pathway interactions; Patients; Pattern; Physiological; Platelet-Derived Growth Factor; Platelet-Derived Growth Factor Receptor; Platelet-Derived Growth Factor alpha Receptor; Platelet-Derived Growth Factor beta Receptor; Play; Population; Process; Proteins; Proto-Oncogene Proteins c-sis; Pulmonary Hypertension; Pulmonary artery structure; Receptor Signaling; Reporter; Research Personnel; Rodent Model; Role; Seminal; Signal Pathway; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Staging; Staining method; Stains; Structure; Sum; TimeLine; Tissues; Transgenes; Transgenic Mice; Tube; Vascular Diseases; Work; cell motility; day; insight; lung development; programs; receptor; receptor expression; research study; restenosis; transcription factor; vascular bed; vascular smooth muscle cell migration

DESCRIPTION (provided by applicant): Dedifferentiation and enhanced proliferation and motility of vascular smooth muscle cells (VSMCs) are key elements in the pathogenesis of many vascular diseases. Thus, a detailed understanding of the development of the VSMC layer of normal blood vessels promises to provide critical insights into vascular disease. However, the processes underlying the morphogenesis of any specific vessel or vascular bed are not well understood. The central goal of this proposal is to elucidate the key molecular and cellular events encompassing the morphogenesis of the smooth muscle cell (SMC) layer of the left pulmonary artery during the initial four days of murine lung development. This goal is the first step towards the long term objective of delineating the critical events in the building of all layers of the pulmonary artery throughout development. An essential element of achieving these goals is to characterize the roles specific signaling pathways play in pulmonary artery development. Platelet derived growth factor (PDGF)-induced signaling has been implicated in the pathogenesis of pulmonary artery hypertension and restenosis following coronary artery angioplasty; yet, the specific effects of PDGF signaling in these diseases or in the morphogenesis of large vessels remain elusive. I hypothesize that signaling through the PDGF pathway is critical for the maintenance, proliferation and/or recruitment of a multipoint early lung mesenchymal cell population that has the capacity to differentiate into the SMC layer of the left pulmonary artery. The proposal has three specific aims: 1) establish a timeline of molecular markers and cellular events that identify specific stages during the transition of a murine lung mesenchymal cell into a differentiated left pulmonary artery SMC and relate this timeline to'endothelial cell and non-vascular tissue development; 2) identify the pattern and effects of activation of PDGF receptor-mediated signaling pathways in left pulmonary artery SMC development; 3) determine the fate in the lung of early PDGF receptor-beta-positive mesenchymal cells through lineage analysis. To achieve these aims, lungs will be dissected from wildtype and transgenic mouse embryos and subjected to whole mount immunostaining, in situ hybridization or X-gal staining. In selected experiments, embryonic lungs will be cultured prior to analysis. Confocal microscopy will be used to analyze fluorescent stains on a cellular level. In sum, the proposed work will make the left pulmonary artery the best understood example of morphogenesis of a specific blood vessel. These studies will yield critical insights into the mechanisms underlying prevalent vascular diseases such as coronary artery atherosclerosis. In addition, this work promises to advance the investigation of critically understudied disorders of smooth muscle cell physiology in the lung, such as pulmonary artery hypertension and lymphangioleiomyomatosis.

描述（由申请人提供）：血管平滑肌细胞（VSMC）的去分化和增强的增殖和运动是许多血管疾病发病机制的关键因素。因此，详细了解正常血管VSMC层的发展有望为血管疾病提供重要的见解。然而，任何特定的血管或血管床的形态发生的过程还没有很好的理解。该建议的中心目标是阐明关键的分子和细胞事件，包括左肺动脉平滑肌细胞（SMC）层的形态发生在小鼠肺发育的最初四天。这一目标是实现长期目标的第一步，即在整个开发过程中描绘肺动脉各层构建中的关键事件。 实现这些目标的一个基本要素是表征特定信号通路在肺动脉发育中的作用。血小板衍生生长因子（PDGF）诱导的信号转导与肺动脉高压和冠状动脉成形术后再狭窄的发病机制有关;然而，PDGF信号转导在这些疾病或大血管形态发生中的具体作用仍然难以捉摸。我推测，通过PDGF途径的信号传导对于多点早期肺间充质细胞群的维持、增殖和/或募集至关重要，该细胞群具有分化成左肺动脉SMC层的能力。 该建议有三个具体目标：1）建立分子标记物和细胞事件的时间轴，其鉴定鼠肺间充质细胞向分化的左肺动脉SMC转变期间的特定阶段，并将该时间轴与内皮细胞和非血管组织发育相关联; 2）鉴定左肺动脉SMC发育中PDGF受体介导的信号传导途径的激活模式和作用; 3）通过谱系分析确定早期PDGF受体-β-阳性间充质细胞在肺中的命运。为了实现这些目标，将从野生型和转基因小鼠胚胎中解剖肺，并进行整体包埋免疫染色、原位杂交或X-gal染色。在选定的实验中，将在分析前培养胚胎肺。共聚焦显微镜将用于在细胞水平上分析荧光染色。 总之，所提出的工作将使左肺动脉成为特定血管形态发生的最好理解的例子。这些研究将产生关键的洞察机制的普遍血管疾病，如冠状动脉粥样硬化。此外，这项工作有望推进研究严重不足的疾病，如肺动脉高压和淋巴管平滑肌瘤在肺平滑肌细胞生理学。