Regulation of coronary vascular development by differential tissue hypoxia

组织缺氧差异对冠状血管发育的调节

• 批准号: 7915293
• 负责人: MICHIKO WATANABE
• 金额: $ 47.83万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7915293
• 关键词: Adult; Apoptosis; Appearance; Architecture; Behavior; Biological Assay; Birds; Blood Vessels; Cardiac; Cells; Child; Clinical; Complex; Coronary; Coronary Vessels; Coronary artery; Defect; Development; Differentiation Antigens; Dominant-Negative Mutation; Embryo; Embryonic Heart; Ensure; Epicardium; Epithelial; Genes; Heart; Hypoxia; Hypoxia Inducible Factor; In Vitro; Infant Mortality; Lead; Location; Mediating; Mesenchymal; Methods; Microarray Analysis; Minor; Modeling; Molecular; Morbidity - disease rate; Myocardial; Myocardium; Nephroblastoma; Nuclear; Pathway interactions; Pattern; Proteins; Pulmonary artery structure; Regulation; Site; Staging; Stem cells; Stereotyping; Sudden Death; System; Thromboplastin; Time; Tissues; Transcriptional Activation; Transcriptional Regulation; Transplanted tissue; Tumor Suppressor Genes; Variant; Viral Vector; chromatin immunoprecipitation; clinically significant; gene therapy; in vivo; left coronary artery; novel; precursor cell; prevent; public health relevance; reconstruction; vasculogenesis

DESCRIPTION (provided by applicant): The major coronary arteries form in stereotyped locations. Abnormalities in location such as in Anomalous Left Coronary Artery from the Pulmonary Artery (ALCAPA) have serious clinical consequences including sudden death. The overall aim of this study is to delineate the mechanisms that guide coronary vessel differentiation and organization. We detected differentially high levels of hypoxia and hypoxia inducible factor -1a (HIF-1a) nuclear localization in the developing heart at specific sites where major coronary vessels will develop. Our hypothesis is that these differential levels of hypoxia within the embryonic heart are critical for steps in coronary vessel development through HIF-1a transcriptional regulation of downstream genes. Our approach will be to characterize the consequences of altering HIF-1a-mediated transcriptional activity by forcing the expression of dominant negative or constitutively active HIF-1a specifically in the myocardium or the epicardium of avian embryos in ovo using viral vectors. We will disrupt HIF-1a activity and assay for consequences to the architecture of the coronary arteries at late stages in development. Disruption of normal vascular patterns will be assessed at different developmental stages using advanced quantitative methods that may allow us to predict which downstream factors are involved. This complex analysis will require 3- D reconstruction and novel quantitative analyses provided by our collaborators. We will next focus on the effects of altering HIF-1a activity on the behavior of coronary precursor cells originating from the proepicardium and epicardium using in vitro systems as well as avian embryos in ovo. We will assay for expression of differentiation markers, epithelial mesenchymal transition, proliferation, apoptosis and myocardial invasion. The last specific aim will be to determine how HIF-1a regulates the activity of the Wilms' tumor suppressor gene product Wt1 in epicardial cells using in vitro systems. Genes directly regulated by HIF-1a and/or Wt1 will be identified. These studies will elucidate how microenvironmental hypoxia, coordinates where and when the major coronary vessels develop. PUBLIC HEALTH RELEVANCE: The clinical significance of this study is that it will increase understanding of coronary vascular anomalies that impact the morbidity and mortality of infants, children, and adults. The findings may serve not only to predict defects earlier, but to lead to strategies to prevent defects. The broader significance of these studies is that they may also lead to strategies for the controlled revascularization of diseased cardiac tissues, transplanted tissues, or newly formed tissues from stem cell or gene therapy.

描述(申请人提供)：主要冠状动脉形成在固定的位置。位置异常，如左冠状动脉起源于肺动脉(ALCAPA)，会导致严重的临床后果，包括猝死。这项研究的总体目标是描绘引导冠状动脉分化和组织的机制。我们在发育中的心脏中发现了不同程度的高水平的低氧和低氧诱导因子-1a(HIF-1a)的核定位于主要冠状动脉发育的特定部位。我们的假设是，胚胎心脏内的这些不同程度的低氧对于通过HIF-1a转录调控下游基因而在冠脉血管发育中的步骤至关重要。我们的方法将描述改变HIF-1a介导的转录活性的后果，方法是使用病毒载体迫使显性阴性或结构性活性的HIF-1a在禽类胚胎的心肌或心外膜中特异地表达。我们将干扰HIF-1a的活性，并分析其对发育后期冠状动脉结构的影响。正常血管模式的破坏将在不同的发育阶段使用先进的定量方法进行评估，这些方法可能使我们能够预测哪些下游因素参与其中。这种复杂的分析将需要我们的合作者提供的3D重建和新颖的定量分析。接下来，我们将利用体外系统和禽类胚胎，重点研究改变HIF-1a活性对起源于心前膜和心外膜的冠状动脉前体细胞行为的影响。我们将检测分化标记物的表达、上皮间充质转化、增殖、凋亡和心肌侵袭。最后一个具体目标将是利用体外系统确定HIF-1a如何调节Wilms的肿瘤抑制基因产物Wt1在体外培养的心外膜细胞中的活性。直接受HIF-1a和/或Wt1调控的基因将被识别出来。这些研究将阐明微环境缺氧是如何协调主要冠状动脉血管何时何地发展的。公共卫生相关性：这项研究的临床意义在于，它将增加对影响婴儿、儿童和成人发病率和死亡率的冠状动脉血管异常的了解。这些发现可能不仅有助于更早地预测缺陷，还可能导致预防缺陷的策略。这些研究的更广泛的意义在于，它们还可能导致对患病的心脏组织、移植组织或干细胞或基因疗法新形成的组织进行受控血运重建的策略。

项目成果

Hypoxia Supports Epicardial Cell Differentiation in Vascular Smooth Muscle Cells through the Activation of the TGFβ Pathway.

• DOI: 10.3390/jcdd5020019
• 发表时间: 2018-04-13
• 期刊: Journal of cardiovascular development and disease
• 影响因子: 2.4
• 作者: Tao J;Barnett JV;Watanabe M;Ramírez-Bergeron D
• 通讯作者: Ramírez-Bergeron D

Epicardial HIF signaling regulates vascular precursor cell invasion into the myocardium.

• DOI: 10.1016/j.ydbio.2013.01.026
• 发表时间: 2013-04-15
• 期刊: DEVELOPMENTAL BIOLOGY
• 影响因子: 2.7
• 作者: Tao, Jiayi;Doughman, Yongqiu;Yang, Ke;Ramirez-Bergeron, Diana;Watanabe, Michiko
• 通讯作者: Watanabe, Michiko