Neurovascular cross-talk, functions and sources of soluble VEGF receptors in ocular vascular demarcation

项目摘要 视觉清晰度取决于眼血管床和视觉关键组织之间严格的血管分界， 角膜和黄斑光感受器。我们的长期目标是阐明维持这一点的分子网络 我们的目的是研究眼部的区室化，并了解病理性眼部血管生成中网络是如何改变的。我们 目前的目标是解剖血管与神经元对血管内皮细胞需求的组织特异性差异， 在正常和病理条件下的生长因子（VEGF）。 我们将确定可溶性血管内皮生长因子受体如何协调角膜间室的相互作用，以滴定平衡 血管分界和神经元稳态的VEGF需求之间的关系。我们将从基因上剖析 特定的眼部隔室，以确定VEGF依赖性神经再生是如何完成的，而不触发 血管破裂我们将严格分析可溶性VEGFR 1（sFlt 1）敲低的血管反应， 上皮伤口愈合和神经再生过程中的角膜隔室。为了补充这一分析，我们将剖析 sVEGF受体在角膜神经元表达中的作用。最后，我们将阐明剪接移位吗啉是否 与靶向新血管的肽基序缀合的药物可以治疗角膜新生血管形成（KNV）和脉络膜新生血管形成。 新生血管形成（CNV）。我们下一个资助期的具体目标是： 具体目标#1：严格测试可溶性VEGFR 1（sFlt 1）或可溶性VEGFR 2（sKDR）的功能要求， 在正常和受伤的角膜中实现血管分界和适当的神经支配的特定角膜隔室 角膜 具体目标#2：确定由Pax 6单倍不足引起的无虹膜相关KNV是否来源于 角膜基质中神经嵴相关的缺陷改变了血管生成的潜力，以及血管是否从角膜缘破裂 干细胞缺乏是由于上皮细胞上调MT 2-MMP引起的次级旁分泌机制。 具体目的#3：确定局部施用RGD缀合的剪接移位吗啉代 （cRGD.KDR.MO）可显著增加激光-CNV损伤中sKDR/mKDR比值，减少新生血管形成， 野生型小鼠的缝合角膜和Pax 6 +/-小鼠角膜。 这些研究将确定sFlt 1和sKDR的组织特异性功能， 角膜中神经元和血管的稳态，系统地详细说明了血管的发展和维持， 正常和病理情况下的分区和神经突生长。

Project Summary Visual clarity depends on strict vascular demarcations between ocular vascular beds and vision-critical tissues such as the cornea and macular photoreceptors. Our long-range goals are to elucidate the molecular network that maintains this compartmentalization in the eye and to understand how the network is altered in pathological ocular angiogenesis. Our immediate goal is to dissect tissue-specific differences in vascular versus neuronal requirements for vascular endothelial growth factor (VEGF) under normal and pathological conditions. We will determine how soluble VEGF receptors orchestrate cross-talk among corneal compartments to titrate the balance between VEGF requirements for vascular demarcation and neuronal homeostasis. We will genetically dissect the roles of specific ocular compartments to determine how VEGF-dependent nerve regeneration is accomplished without triggering a vascular breach. We will rigorously analyze the vascular response of soluble VEGFR1 (sFlt1) knockdown within each corneal compartment during epithelial wound healing and nerve regeneration. Complementing this analysis, we will dissect the roles of sVEGF receptors expressed on corneal neurons. Finally, we will elucidate whether splice shifting morpholinos conjugated to a neovessel-targeting peptide motif can treat corneal neovascularization (KNV) and choroidal neovascularization (CNV). Our specific aims for the next grant period are: Specific Aim #1: To rigorously test functional requirements for soluble VEGFR1 (sFlt1) or soluble VEGFR2 (sKDR) within specific corneal compartments for achieving vascular demarcation and appropriate innervation in the normal and injured cornea. Specific Aim #2: To determine whether aniridia-associated KNV resulting from Pax6 haploinsufficiency derives from neural crest related defects in the corneal stroma that shift angiogenic potential, and whether vascular breach from limbal stem cell deficiency is a secondary paracrine mechanism due to epithelial upregulation of MT2-MMP. Specific Aim #3: To determine whether local administration of RGD-conjugated splice shifting morpholino (cRGD.KDR.MO) can significantly increase sKDR/mKDR ratios and decrease neovascularization in laser-CNV injury and sutured corneas of wild type mice, and in Pax6+/- mouse corneas. These studies will define the tissue-specific functions of sFlt1 and sKDR in the context of competing requirements for neuronal and vascular homeostasis in the cornea, systematically detailing the development and maintenance of vascular zoning and neurite outgrowth in normal and pathological situations.