Integrating Cell Division and Morphogenesis in Developing Vessels

描述(申请人提供)：为了制造一个合适的血管，几个主要的细胞过程必须被调节和整合。具体地说，内皮细胞分裂发生在导致血管网络萌芽形成、融合和扩张的形态发生过程中。正常情况下，这些不同的过程被优雅地交织在一起，以产生适量的具有适当三维图案的血管。然而，关于血管生成过程中内皮细胞分裂和形态发生在空间和时间上是如何调控的，人们知之甚少，更不知道这些过程是如何整合形成血管的。我们有证据表明，内皮细胞分裂的两个主要方面受到形态发生线索的调节，即细胞分裂的速度和有丝分裂期间裂解平面的方向。因此，我们假设形态发生信号影响内皮细胞分裂的特定参数，并且这种输入对正确的血管形态发生至关重要。我们还假设，影响内皮细胞分裂速度和方向的形态发生信号是通过内皮细胞-细胞连接和有丝分裂极性成分传递的。为了验证这些假说，我们将使用动态成像来阐明血管生成过程中内皮细胞分裂在时间和空间上受到调节的“规则”。我们将操纵影响纺锤体动力学的内皮连接和极性分子，并确定这些操纵对细胞分裂和极性的影响。最后，我们将详细研究两个信号通路，血管内皮生长因子和平面细胞极性(PCP或非规范Wnt信号)在内皮细胞形态发生和细胞分裂之间的协调中所起的作用。对血管生成过程中细胞过程如何整合的分子理解将有助于设计血管再生的方法。能够概括生物过程导致适当的血管形成是再生医学的许多方面的要求，因此这项工作将在这个翻译领域产生很大影响。这项提议将使用老鼠模型来研究构成血管壁的细胞的繁殖如何影响血管的形状和形状。结果将帮助我们了解血管是如何形成的，这反过来将有助于设计制造用于治疗的人造血管的方法。

DESCRIPTION (provided by applicant): To make a proper blood vessel, several major cellular processes must be regulated and integrated. Specifically, endothelial cell division occurs in the context of morphogenetic processes that lead to sprout formation, fusion, and expansion of the vascular network. Normally these distinct processes are elegantly interwoven to produce the appropriate amount of vasculature with the proper 3-dimensional pattern. However, relatively little is known about how endothelial cell division and morphogenesis are regulated in space and time during angiogenesis, and even less is known about how these processes integrate to form blood vessels. We have evidence that two major aspects of endothelial cell division are regulated by morphogenetic cues, the rate of cell division and the orientation of the cleavage plane during mitosis. Thus we hypothesize that morphogenetic signals impact specific parameters of endothelial cell division, and that this input is critical to proper vessel morphogenesis. We also hypothesize that morphogenetic signals affecting the rate and orientation of endothelial cell division are transduced via endothelial cell-cell junctions and mitotic polarity components. To test these hypotheses, we will use dynamic imaging to elucidate the "rules" by which endothelial cell division is regulated in time and space during angiogenesis. We will manipulate endothelial junctions and polarity molecules that affect spindle dynamics, and determine the impact of these manipulations on cell division and polarity. Finally, we will examine in detail the role of two signaling pathways, VEGF and Planar Cell Polarity (PCP or non-canonical Wnt signaling) in the co- ordination between endothelial morphogenesis and cell division. A molecular understanding of how cellular processes are integrated during angiogenesis will help in the design of approaches to vessel regeneration. The ability to recapitulate biological processes leading to proper vessel formation is a requirement for many aspects of regenerative medicine, so this work will have high impact in this translational arena. This proposal will use mouse models to examine how the propagation of cells making up the wall of blood vessels affects the form and shape of the vessel. The results will help us understand how vessels are shaped, which in turn will aid in designing ways to make artificial vessels for therapeutic use.