Flt-1 (VEGFR-1) Regulation of Endothelial Cell Sprouting and Vessel Morphogenesis

Flt-1 (VEGFR-1) 调节内皮细胞出芽和血管形态发生

基本信息

批准号：
7614747
负责人：
John Christopher Chappell
金额：
$ 4.96万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-01-01 至 2010-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7614747
关键词：
Affinity Binding Blood Vessels Cell Culture Techniques Cell Survival Cell division Cell model Cells Complex Coronary heart disease Cues Development Diabetic Retinopathy Disease Embryo Embryonic Development Endothelial Cells Filopodia Generations Genetic Goals Growth Growth Factor Growth and Development function Knowledge Ligands Link Location Mechanics Modeling Modification Molecular Morphogenesis Morphology Mus Nature Notch Signaling Pathway Pathway interactions Patients Pattern Peripheral Vascular Diseases Play Process Protein Isoforms Protein Tyrosine Kinase Regulation Relative (related person)Retina Role Signal Transduction Spatial Distribution Testing Tissues Vascular Endothelial Growth Factor A Vascular Endothelial Growth Factor Receptor-1 angiogenesis clinically relevant design embryonic stem cell inhibitor/antagonist insight migration mutant neutralizing antibody notch protein receptor receptor binding retina blood vessel structure secretase stem therapeutic development therapy design tumor growth vasculogenesis

项目摘要

DESCRIPTION (provided by applicant): Vascular networks develop through several process including vasculogenesis, the de novo formation of primitive vessels, and angiogenesis, the sprouting of new vessels from pre-existing ones. Vascular endothelial growth factor-A (VEGF) and its receptors, Flt-1 and Flk-1, play critical roles in controlling these processes. VEGF signaling effects via Flk-1 have been well characterized, but the role of Flt-1 in regulating vessel formation remains unclear. Flt-1 regulates vessel branching, primarily by producing a soluble Flt-1 isoform that acts largely as a ligand sink. Furthermore, recent evidence suggests that Notch signaling interacts with VEGF pathways to guide developing vessels, although the exact nature of this interaction is not well understood. Thus, the hypotheses that will be tested are that Flt-1 regulates vessel morphogenesis by providing spatial cues to guide endothelial cell (EC) filopodia and sprout formation, and that Notch signaling modulates downstream Flt-1 activity to establish the proper pattern of sprout extension and in turn vessel branching. To achieve these goals, models of vessel formation in differentiated embryonic stem (ES) cell cultures and in mouse retina development will be utilized. In both models, VEGF signaling will be disrupted through genetic modification of Flt-1 expression. After characterizing the spatial distribution of Flt- 1 expression in these models, Flt-1 mutant and wild-type (WT) vessels will be compared with regard to EC filopodia extension, tip cell formation, vessel sprout guidance, and overall vessel branching and morphology. To characterize how Notch signaling regulates Flt-1 in guiding vessel sprouts, the location of Notch signaling relative to EC filopodia and vessel sprouts will be established in both the WT and Flt-1 mutant ES cell and retina vessel models. Notch signaling will be disrupted genetically and through the use of inhibitors or neutralizing antibodies. Developing vessels will be analyzed as described above. From these studies, knowledge will be gained regarding how Flt-1 regulates VEGF signaling such that proper EC filopodia extension, sprout formation and vessel branching occur, and how Notch signaling coordinates Flt-1 activity to yield properly branched vascular networks. Insights from this study will be essential in the development of therapeutic strategies to disrupt blood vessel growth in treating certain pathological conditions such as tumor growth and diabetic retinopathy. Furthermore, this study will be informative in the design of clinically relevant treatments for stimulating blood vessel formation in patients suffering from vascular occlusive diseases such as coronary heart disease and peripheral vascular disease.

描述（由申请人提供）：血管网络通过几个过程发育，包括血管生成、原始血管的从头形成和血管生成、新血管从先前存在的血管中萌芽。血管内皮生长因子-A (VEGF) 及其受体 Flt-1 和 Flk-1 在控制这些过程中发挥着关键作用。通过 Flk-1 的 VEGF 信号传导作用已得到很好的表征，但 Flt-1 在调节血管形成中的作用仍不清楚。 Flt-1 主要通过产生可溶性 Flt-1 亚型来调节血管分支，该亚型主要充当配体汇。此外，最近的证据表明，Notch 信号传导与 VEGF 通路相互作用以引导血管发育，尽管这种相互作用的确切性质尚不清楚。因此，将要测试的假设是 Flt-1 通过提供空间线索来引导内皮细胞 (EC) 丝状伪足和芽形成来调节血管形态发生，并且 Notch 信号传导调节下游 Flt-1 活性以建立正确的芽延伸模式进而血管分支。为了实现这些目标，将利用分化胚胎干（ES）细胞培养物和小鼠视网膜发育中的血管形成模型。在这两种模型中，VEGF 信号传导将通过 Flt-1 表达的基因修饰而被破坏。在表征这些模型中 Flt-1 表达的空间分布后，将比较 Flt-1 突变体和野生型 (WT) 血管的 EC 丝状伪足延伸、尖端细胞形成、血管萌芽引导以及整体血管分支和形态。为了表征Notch信号如何在引导血管芽中调节Flt-1，将在WT和Flt-1突变体ES细胞和视网膜血管模型中建立Notch信号相对于EC丝状伪足和血管芽的位置。 Notch 信号传导将通过基因和抑制剂或中和抗体的使用而被破坏。开发中的船只将如上所述进行分析。从这些研究中，我们将了解 Flt-1 如何调节 VEGF 信号传导，从而实现适当的 EC 丝状伪足延伸、芽形成和血管分支，以及 Notch 信号传导如何协调 Flt-1 活性以产生适当分支的血管网络。这项研究的见解对于制定破坏血管生长的治疗策略至关重要，以治疗某些病理状况，例如肿瘤生长和糖尿病视网膜病变。此外，这项研究将为刺激患有血管闭塞性疾病（例如冠心病和外周血管疾病）患者的血管形成的临床相关治疗的设计提供信息。