VEGF-C/VEGFR3 Regulation of VE-Cadherin in Sinusoidal Angiogenesis and Lymphangiogenesis

VEGF-C/VEGFR3 对 VE-钙粘蛋白在正弦血管生成和淋巴管生成中的调节

• 批准号: 10563110
• 负责人: Derek C Sung
• 金额: $ 3.41万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10563110
• 关键词: Ablation; Binding; Blood; Blood Vessels; Bone Marrow; Cell Culture Techniques; Cells; Cre lox recombination system; Data; Defect; Development; Discontinuous Capillary; Edema; Electrical Resistance; Embryo; Embryonic Development; Endocytosis; Endothelial Cells; Endothelium; Erythroid; Fetal Liver; Fluid Balance; Genetic; Goals; Growth; Growth and Development function; Hematopoiesis; Histologic; Human; Immunofluorescence Immunologic; In Vitro; KDR gene; Knockout Mice; Ligands; Liver; Liver Regeneration; Lymphangiogenesis; Lymphatic; Lymphatic Diseases; Lymphatic Endothelial Cells; Measures; Mediating; Membrane; Modeling; Molecular; Mus; Organ; Pathway interactions; Permeability; Pharmacology; Phenocopy; Phenotype; Phosphorylation; Process; Regulation; Research; Role; Signal Transduction; Signaling Molecule; Skin; Small Interfering RNA; Stains; Testing; Tissues; VEGFA gene; Vascular Endothelial Growth Factor C; Vascular Endothelial Growth Factor Receptor-3; Vascular Endothelial Growth Factors; Western Blotting; Work; angiogenesis; beta-arrestin; blood vessel development; cadherin 5; cell growth; gain of function; improved; in vivo; inhibitor; insight; lymphatic malformations; lymphatic vasculature; novel; reconstitution; regenerative therapy; therapeutic target; vascular bed

Research Summary The blood, lymphatic, and sinusoidal vasculatures are molecularly and functionally distinct vascular networks whose normal growth and development are essential to support embryogenesis. While there has been notable effort to understand mechanisms of blood vessel development, relatively little is known about the role of vascular endothelial growth factor (VEGF) receptor 3 (VEGFR3) and its main ligand VEGF-C in sinusoidal development. Despite decades of work on VEGFR2 and VEGFR3 signaling, insights into the field have been complicated by use of various genetic and pharmacologic models with divergent phenotypes. Furthermore, VEGF-C can weakly bind and activate VEGFR2 in addition to inducing VEGFR2-VEGFR3 heterodimers. As a result, the role of VEGF-C in vascular development remains incompletely understood. Therefore, our goal is to determine the role of VEGF-C/VEGFR3 in vascular development and identify novel signaling mechanisms through which it functions through a combination of in vivo mouse and in vitro cell culture studies. We and others have previously identified VEGF-C as an important regulator of fetal liver hematopoiesis. This was primarily attributed to defects in erythroid maturation, however the sinusoids are the only VEGFR3-positive cells in the fetal liver. Strikingly, endothelial deletion of VEGFR3 and vascular endothelial (VE-)cadherin gain- of-function mice both phenocopy VEGF-C knockout mice, suggesting that diminished fetal liver hematopoiesis is in fact due to sinusoidal defects. These mice also have decreased bone marrow (BM) sinusoids. BM defects due to endothelial loss of VEGFR3 are rescued by VE-cadherin haploinsufficiency, demonstrating that VEGFR3 negatively regulates VE-cadherin to promote sinusoidal growth. Whether this process depends on VEGF-C and the signaling mechanisms through which VEGFR3 regulates VE-cadherin remain unclear. Therefore, I hypothesize that VEGF-C activates VEGFR3 to directly and negatively regulate VE-cadherin during sinusoidal angiogenesis and lymphangiogenesis. Aim 1 will test whether haploinsufficiency of VE- cadherin can rescue fetal liver and lymphatic growth defects, as I have demonstrated to be true in the bone marrow sinusoids. Aim 2 will test whether VEGFR3 functions in a VEGF-C-dependent or independent manner in the skin, liver, and bone marrow during sinusoidal angiogenesis and lymphangiogenesis. Finally, Aim 3 will determine the mechanism by which VEGFR3 negatively regulates VE-cadherin through in vitro studies using human lymphatic endothelial cells. Together, these studies will bring novel insights into the mechanism by which VEGF-C/VEGFR3 may regulate VE-cadherin across various vascular beds in multiple organs, which could have important implications for liver regeneration, BM reconstitution, and lymphatic disorders.

研究摘要 血液，淋巴和正弦血管是分子和功能上不同的血管网络 其正常的生长和发育对于支持胚胎发生至关重要。虽然有著名的 努力了解血管发育的机制，对 血管内皮生长因子（VEGF）受体3（VEGFR3）及其主要配体vegf-c在正弦中 发展。尽管在VEGFR2和VEGFR3信号上进行了数十年的工作，但对该领域的见解一直是 通过使用各种具有不同表型的遗传和药理学模型来复杂。此外， VEGF-C除了诱导VEGFR2-VEGFR3异二聚体外，还可以弱结合和激活VEGFR2。作为 结果，VEGF-C在血管发育中的作用尚未完全理解。因此，我们的目标是 确定VEGF-C/VEGFR3在血管发育中的作用并确定新型信号传导机制 通过体内小鼠和体外细胞培养研究的结合，它通过它发挥作用。我们和 其他人先前已将VEGF-C确定为胎儿肝造血的重要调节剂。这是 主要归因于红斑成熟的缺陷，但是正弦是唯一的VEGFR3阳性 胎儿肝脏中的细胞。引人注目的是，VEGFR3和血管内皮（VE-）钙粘蛋白增益的内皮缺失 - 功能小鼠两种表观复制VEGF-C基因敲除小鼠，表明胎儿肝造血减少 实际上是由于正弦缺陷。这些小鼠还减少了骨髓（BM）正弦曲线。 BM缺陷 由于内皮的损失，VEGFR3被VE-钙粘着蛋白的单倍症挽救了，表明 VEGFR3负调节VE-钙粘蛋白以促进正弦生长。这个过程是否取决于 VEGF-C和VEGFR3调节VE-Cadherin的信号传导机制尚不清楚。 因此，我假设VEGF-C激活VEGFR3直接和负调节VE-Cadherin 在正弦血管生成和淋巴管生成期间。 AIM 1将测试Ve-ve-ve- 钙粘蛋白可以挽救胎儿肝脏和淋巴生长缺陷，正如我在骨骼中是真实的那样 骨髓正弦。 AIM 2将测试VEGFR3是以VEGF-C依赖性还是独立的方式运行 在正弦血管生成和淋巴管生成期间的皮肤，肝脏和骨髓中。最后，AIM 3将 确定VEGFR3通过使用体外研究对VEGFR3负调节VE-钙黏着蛋白的机制 人淋巴内皮细胞。这些研究将通过 VEGF-C/VEGFR3可以调节多个器官中各种血管床的VE-钙粘蛋白 可能对肝脏再生，BM重建和淋巴性疾病具有重要意义。