Anti-angiogenic activity of neurokinin B

性状（由申请方提供）：神经激肽B（NK-B）是速激肽家族的一种肽。经典地，在神经系统的背景下研究NK-B表达和功能。然而，NK-B的过表达与妊娠相关疾病先兆子痫有关，先兆子痫的特征在于胎盘血管系统的缺陷性重塑和全身血管失调。我们证明，NK-B可逆地抑制内皮细胞血管网络组装在体外基质胶和反对血管生成在体内。机制分析表明，NK-B信号抑制血管生成信号诱导的血管内皮生长因子受体1（VEGFR 1）和VEGFR 2的表达，并减少内皮细胞迁移。本提案的目的是确定NK-B抑制血管生成信号诱导的VEGFR 1和VEGFR 2诱导以及内皮细胞迁移的调节机制。我们将制定两个具体目标来解决这个问题。在第一个具体目标中，我们将确定神经激肽B如何抑制血管生成信号依赖性VEGFR诱导。转录因子Ets 1和HIF-2 α参与VEGFR 1和VEGFR 2的转录调控。我们将测试Ets 1和HIF-2 α是否在体外和体内直接激活VEGFR 1和VEGFR 2的转录以响应血管生成信号，以及NK-B信号是否通过阻断Ets 1/HIF-2 α染色质占用或占用后的关键功能来抑制转录。在第二个具体目标中，我们将确定NK-B介导的内皮细胞迁移抑制的机制。我们假设，在内皮细胞中，NK-B信号调节介导定向细胞迁移的关键信号通路。为了验证这一假设，我们将记录刺激内皮细胞进行迁移的信号转导事件和NK-B对这些事件的影响，包括粘着斑激酶，桩蛋白，p130 Cas的磷酸化，这些分子的空间和时间关联形成粘着斑，以及GTP酶Rac 1和CDC 42的Rho家族的激活。公共卫生相关性：血管生成，即从现有血管系统发育出新血管，是许多生理过程中的关键事件，如器官生长和发育、伤口愈合和生殖。血管生成对于包括肿瘤生长/转移的某些病理学病症也是关键的。从这些研究中收集的信息将有助于我们了解血管生成的分子机制，并有望导致新的治疗方式。

DESCRIPTION (provided by applicant): Neurokinin B (NK-B) is a member of tachykinin family of peptides. Classically, NK-B expression and function was studied in the context of the nervous system. However overexpression of NK-B has been implicated in the pregnancy-associated disorder preeclampsia, which is characterized by defective remodeling of the placental vasculature and systemic vascular deregulation. We demonstrated that NK-B reversibly inhibits endothelial cell vascular network assembly in vitro on Matrigel and opposes angiogenesis in vivo. Mechanistic analyses revealed that NK-B signaling inhibited angiogenic signal-induced expression of vascular endothelial growth factor receptor1 (VEGFR1) and VEGFR2, and reduced endothelial cell migration. The objective of this proposal is to determine the regulatory mechanisms, by which NK-B inhibits angiogenic signal-induced VEGFR1 and VEGFR2 induction, and endothelial cell migration. We will institute two specific aims to address this. In the first Specific Aim we will determine how neurokinin B represses angiogenic signal-dependent VEGFR induction. Transcription factors Ets1 and HIF-2alpha have been implicated in the transcriptional regulation of VEGFR1 and VEGFR2. We will test whether Ets1 and HIF-2alpha directly activate transcription of VEGFR1 and VEGFR2 in response to angiogenic signals in vitro and in vivo, and whether NK-B signaling represses transcription by blocking Ets1/HIF-2alpha chromatin occupancy or a critical function post-occupancy. In the second Specific Aim we will determine the mechanism of NK-B-mediated inhibition of endothelial cell migration. We hypothesize that, in endothelial cells, NK-B signaling modulates critical signaling pathways that mediate directed cell migration. To test this hypothesis we will document signal transduction events in endothelial cells stimulated to undergo migration and the effect of NK-B on those events, including phosphorylation of focal adhesion kinase, paxillin, p130Cas, spatial and temporal association of these molecules to form focal adhesions, and activation of Rho family of GTPases Rac1 and CDC42. PUBLIC HEALTH RELEVANCE: Angiogenesis, the development of new blood vessels from existing vasculature, is a key event in many physiological processes, like organ growth and development, wound healing, and reproduction. Angiogenesis is also critical for certain pathological disorders including tumor growth/metastasis. Information gleaned from these studies will contribute to our understanding of the molecular mechanisms of angiogenesis and hopefully will lead to novel therapeutic modalities.