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力关键组织之间严格的血管分界，例如 角膜和黄斑光感受器。我们的长期目标是阐明维持这种状态的分子网络 目的是为了了解眼部神经网络在病理性眼血管生成中是如何改变的。我们的 近期的目标是剖析血管和神经元对血管内皮细胞需求的组织特异性差异 正常和病理条件下的生长因子(VEGF)。 我们将确定可溶性的血管内皮生长因子受体如何协调角膜间隔间的串扰，以滴定平衡。 血管划分的血管内皮生长因子要求和神经元动态平衡之间的关系。我们将从基因上剖析 以确定如何在不触发血管内皮生长因子依赖的神经再生的情况下完成 血管破裂。我们将严格分析每个基因中可溶性VEGFR1(SFlt1)基因敲除的血管反应 角膜上皮损伤愈合和神经再生过程中的角膜隔膜。作为对此分析的补充，我们将剖析 角膜神经元表达sVEGF受体的作用最后，我们将阐明拼接移位吗啉是否 结合neovessel靶向多肽基序可以治疗角膜新生血管(KNV)和脉络膜 新生血管(CNV)。我们在下一个资助期的具体目标是： 具体目标#1：严格测试体内对可溶性VEGFR1(SFlt1)或可溶性VEGFR2(SKDR)的功能需求 在正常和损伤的人中实现血管划分和适当的神经支配的特定角膜隔室 眼角膜。 具体目标2：确定由Pax6单倍体功能不全引起的无虹膜相关性KNV是否源于 角膜基质中与神经脊相关的改变血管生成潜能的缺陷，以及血管是否从角膜缘破裂 干细胞缺乏是一种继发性旁分泌机制，其原因是上皮细胞MT2-MMPs表达上调。 具体目标#3：确定局部应用RGD-连接剪接转移吗啡 (cRGD.KDR.MO)可显著提高激光CNV损伤后sKDR/mKDR比值，减少新生血管形成。 野生型小鼠角膜缝合，Pax6+/-小鼠角膜缝合。 这些研究将在竞争要求的背景下定义sFlt1和sKDR的组织特异性功能 角膜中神经元和血管的动态平衡，系统地详细描述了血管的发育和维持 正常和病理情况下的分区和轴突生长。

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.