Cell lineage induction during coronary vessel development

冠状血管发育过程中的细胞谱系诱导

• 批准号: 6893308
• 负责人: Takashi Mikawa
• 金额: $ 37.79万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6893308
• 关键词: angiogenesis; apoptosis; cell differentiation; cell migration; cell population study; cell type; chick embryo; coronary vessels; developmental genetics; embryogenesis; endothelin; fibroblast growth factor; neural crest; pericardium; transfection

Although functional coronary arterial circulation is vital to cardiac viability and function, very little is known about how the branching pathways of the coronary arterial network are established during embryogenesis Our retroviral cell lineage studies of the chick embryos have demonstrated conclusively that coronary progenitors arises from the epicardial anlagen and that coronary progenitors enter the tubular stage heart with the growing epicardial mantle. In addition, coronary vessels are established by vasculogenic mechanisms and not by angiogenic sprouting form the aorta. We have also shown that coronary vasculogenesis is more prominent in the outer than the inner myocardium and serves as a template for coronary arterial development. This morphogenetic process can be altered by the local expression of exogenous genes (e.g., FGF) using retroviral-mediated gene transfer. Finally, ablation of cardiac neural crests results in the loss of differentiation of the intramural, but not the subepicardial, arterial network. These data lead to the central hypothesis that paracrine signals form the myocardium regulate both the directed migration of coronary vessel precursors to the heart (Hypothesis-1) and capillary plexus formation within the heart (Hypothesis-2). We also hypothesis that a sub-population of neural crest-derived cells regulate the differentiation and patterning of the coronary arterial network (Hypothesis-3). We will test these three specific hypotheses by: 1) determining the region-specific interaction between pro-epicardial cells and myocardium-derived instructive signals and analyzing myocardial signaling properties; 2) reprogramming the gradient of the myocyte-derived FGF within the heart wall by using retroviral vectors expressing FGF-ligands and quantifying alteration of coronary patterning using lineage-specific markers; and 3) retrovirally expressing endothelin, NT3/trkC, or their dominant negative mutants to alter their expression in the developing heart and assaying neural crest migration, differentiation and apoptosis in vivo to determine their requirement for intramural arterial development. The studies proposed here will identify the regulators in establishing coronary artery network and build a foundation for rational therapeutics of coronary disorders in adults.

尽管功能性冠状动脉循环对于心脏的生存力和功能至关重要，但对于在胚胎发生过程中如何建立冠状动脉网络的分支途径如何，我们的雏鸡胚胎逆转录病毒细胞谱系的研究最终证明了冠状动脉祖细胞的逆转录病毒细胞谱系，表明冠状祖细胞是由表皮型胎盘群体产生的。 地幔。此外，冠状动脉血管是通过血管生成机制建立的，而不是通过主动脉形成血管生成的发芽。我们还表明，冠状动脉血管生成在外部心肌中更为突出，并且是冠状动脉发育的模板。使用逆转录病毒介导的基因转移的外源基因（例如FGF）的局部表达可以改变这种形态发生过程。最后，心脏神经rest的消融导致壁内的分化丧失，但没有心膜下动脉网络。这些数据导致了一个中心假设，即旁分泌信号形成心肌既调节心脏（假设1）的冠状动脉血管前体的定向迁移和心脏内毛细血管丛的形成（假设2）。我们还假设神经衍生细胞的亚群调节冠状动脉网络的分化和模式（假设3）。我们将通过以下方式测试这三个特定的假设：1）确定心发细胞与心肌衍生的指导性信号之间的区域特异性相互作用，并分析心肌信号传导特性； 2）通过使用表达FGF-配体的逆转录病毒载体并使用谱系特异性标记来量化冠状动脉模式的改变，对心脏壁内肌细胞衍生的FGF的梯度进行重新编程； 3）逆转录病毒表达内皮素，NT3/TRKC或它们的主要阴性突变体，以改变其在发育中的心脏中的表达，并在体内分化神经rest迁移，分化和凋亡，以确定其对壁内动脉发育的需求。这里提出的研究将确定建立冠状动脉网络的调节剂，并为成人冠状动脉疾病的理性治疗奠定基础。