Molecular Mechanisms Regulating Neuropilin 2 in Lymphangiogenesis

调节淋巴管生成中 Neuropilin 2 的分子机制

• 批准号: 10322716
• 负责人: DIANE Renee BIELENBERG
• 金额: $ 76.38万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-18 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322716
• 关键词: Ablation; Acute; Adult; Animal Model; Attenuated; Binding; Biological Assay; Cell Proliferation; Cell Surface Receptors; Cells; ChIP-seq; Chronic; Coculture Techniques; Complex; Cornea; Data; Disease; Disease model; Dissection; Embryo; Enhancers; Epithelial; Exhibits; FOXC2 gene; Family; Fluid Balance; Fostering; Functional disorder; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Goals; Health; Homeostasis; Immune; Immunologic Surveillance; Immunoprecipitation; In Vitro; Inflammation; Inflammatory; Intestines; Ligands; Limb structure; Lipids; Liquid substance; Luciferases; Lymphangiogenesis; Lymphatic; Lymphatic Diseases; Lymphatic Endothelial Cells; Lymphatic Endothelium; Lymphatic System; Lymphatic function; Lymphedema; Mediating; Medical; Metabolic Diseases; Mission; Modeling; Molecular; Mus; Mutation; Natural regeneration; Near-infrared optical imaging; Neuropilin-2; Operative Surgical Procedures; Pathologic; Patients; Phenotype; Play; Process; Proteins; Public Health; Publications; Reaction; Recycling; Regenerative response; Reporter; Research; Resolution; Role; Semaphorin-3; Semaphorins; Signal Transduction; Surgical sutures; Swelling; Tail; Testing; Therapeutic; Tissues; Transcription Coactivator; Transgenic Animals; Translating; Tube; United States National Institutes of Health; VEGFC gene; Vascular Endothelial Growth Factor C; Vascular Endothelial Growth Factor Receptor-3; Vascular Endothelial Growth Factors; Wound models; apoAI regulatory protein-1; gain of function; improved; in vivo; innovation; keratinocyte; loss of function; lymphatic development; lymphatic drainage; lymphatic dysfunction; lymphatic vessel; member; migration; mouse model; novel; novel strategies; plexin; primary lymphedema; promoter; receptor; regenerative; repaired; secondary lymphedema; targeted treatment; therapeutic target; trafficking; transcription factor; uptake

The lymphatic system is essential in mediating tissue fluid homeostasis, intestinal lipid uptake and immune surveillance. Lymphatic diseases including lymphedema occur from dysfunctional lymphatic networks and are a significant public health issue that may cause loss of function in the limbs. How dysfunctional lymphatics aggravate lymphatic diseases, and conversely, how dysfunctional lymphangiogenesis improves lymphatic disorders are poorly understood but important medically-relevant questions. Our long-term goal is to understand the cellular and molecular processes that control lymphangiogenesis and lymphatic function in order to better treat lymphatic diseases. The Neuropilin 2 (NRP2) receptor is unique in that it transmits both pro-lymphangiogenic signals from VEGF-C/D ligands in concert with VEGFR3 as well as suppressive signals from Semaphorin-3F (SEMA3F) via plexin receptors. Primary lymphedema patients have been found to have mutations in Foxc2, VEGFR3, or NRP2. Our recent publication showing prolonged lymphedema, persistent swelling, and insufficient lymphatic drainage in NRP2-deficient mice during acute inflammatory reactions provides a strong premise for this proposal. Our new mice which inducibly lack NRP2 in lymphatic endothelium have attenuated lymphangiogenesis. Additionally, new data demonstrates that Foxc2 negatively regulates NRP2 (as well as VEGFR3) expression. Together, these data suggest that NRP2 is necessary for normal resolution of extravasated fluid. We hypothesize that the NRP2 signaling axis can be manipulated for therapeutic gain in models of lymphatic dysfunction and chronic inflammation. We will test our hypothesis by pursuing three interrelated yet independent aims. Aim 1 will explore the mechanisms underlying NRP2 and VEGFR3 signaling in adult lymphangiogenesis in vivo using tissue-specific genetic ablation strategies and quantitative near infrared imaging to evaluate lymphatic drainage in the context of inflammation. Given that Foxc2 downregulates NRP2 but NRP2 is normally upregulated during pathological conditions, we will assess NRP2-mediated augmentation of lymphatic function in adult wild type and lymphatic-specific Foxc2 loss-of-function mice. Aim 2 will examine the molecular mechanisms regulating NRP2 expression in lymphatic endothelium. Correspondingly, lymphatic-specific Foxc2 gain-of-function mice exhibit reduced lymphatic function similar to lymphatic- specific NRP2-deficient mice. How NRP2 expression is regulated via Foxc2 is poorly understood but new data suggests that it may interfere with COUP-TFII activity. Aim 3 will investigate the therapeutic potential of inhibiting endogenous SEMA3F to treat lymphatic dysfunctions. Overall, this project is innovative as it will generate novel transgenic animal models, reveal distinct regulatory molecular mechanisms in lymphatic endothelium, and provide original potential therapeutic targets. In summary, our findings could translate into innovative approaches to treat lymphatic dysfunctions including lymphedema.

淋巴系统在调节组织液平衡、肠道脂质摄取和免疫方面是必不可少的。 监视系统。包括淋巴水肿在内的淋巴疾病是由淋巴网络功能障碍和 是一个重大的公共卫生问题，可能会导致肢体功能丧失。功能多么失灵 淋巴管加重淋巴疾病，反之，功能障碍的淋巴管生成如何改善 淋巴疾病虽然知之甚少，但却是与医学相关的重要问题。我们的长期目标 是为了了解控制淋巴管生成和淋巴循环的细胞和分子过程 功能，以便更好地治疗淋巴疾病。神经粘蛋白2(NRP2)受体的独特之处在于它 与VEGFR3协同传递来自VEGF-C/D配体的促淋巴管生成信号以及 Semaphorin-3F(SEMA3F)通过丛状受体抑制信号。原发性淋巴水肿患者 已被发现在FOXC2、VEGFR3或NRP2有突变。我们最近的出版物显示 NRP2基因缺陷小鼠的淋巴水肿、持续性肿胀和淋巴引流不足 在急性炎症反应期间为这一提议提供了强有力的前提。我们的新老鼠 淋巴管内皮细胞中诱导缺失NRP2可抑制淋巴管生成。此外，新数据 表明FOXC2负调控NRP2(以及VEGFR3)的表达。加在一起，这些 数据表明，NRP2是正常分解渗出液所必需的。我们假设 在淋巴功能障碍模型中，NRP2信号轴可以被操纵以获得治疗收益 慢性炎症。我们将通过追求三个相互关联但独立的目标来检验我们的假设。 目的1探讨NRP2和VEGFR3信号在成人淋巴管生成中的作用机制 活体使用组织特异性基因消融策略和定量近红外成像进行评估 炎症背景下的淋巴引流。假设FOXC2下调了NRP2，但NRP2下调了 通常在病理条件下上调，我们将评估NRP2介导的 成年野生型和淋巴特异性FOXC2功能丧失小鼠的淋巴功能。Aim 2将检查 淋巴管内皮细胞NRP2表达调控的分子机制。相应地， 淋巴特异的FOXC2功能获得小鼠表现出与淋巴功能相似的淋巴功能降低。 特定的NRP2缺陷小鼠。NRP2的表达是如何通过FOXC2来调节的还不是很清楚，但这是一个新的现象 数据显示，它可能会干扰政变TFII的活动。目标3将调查治疗潜力 抑制内源性SEMA3F治疗淋巴功能障碍。总的来说，这个项目是创新的，因为它 将产生新的转基因动物模型，揭示淋巴系统不同的调控分子机制 血管内皮细胞，并提供原创的潜在治疗靶点。总而言之，我们的发现可以转化为 进入治疗淋巴功能障碍的创新方法，包括淋巴水肿。