Molecular Mechanisms Regulating Neuropilin 2 in Lymphangiogenesis

调节淋巴管生成中 Neuropilin 2 的分子机制

基本信息

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
项目类别：
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

项目摘要

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.

淋巴系统对于介导组织液体稳态，肠脂质摄取和免疫力至关重要监视。包括淋巴水肿在内的淋巴疾病来自功能失调的淋巴网络和是一个重要的公共卫生问题，可能会导致四肢功能丧失。功能失调淋巴管加剧淋巴病，相反，功能障碍淋巴管生成如何改善淋巴性疾病知之甚少，但重要的医学疾病问题。我们的长期目标是了解控制淋巴管生成和淋巴的细胞和分子过程功能以更好地治疗淋巴病。 Neuropilin 2（NRP2）受体是独一无二的与VEGFR3一起从VEGF-C/D配体中传输促淋巴生殖器信号以及来自Semaphorin-3F（SEMA3F）的抑制信号通过丛受体。原发性淋巴水肿患者已经发现在FOXC2，VEGFR3或NRP2中具有突变。我们最近的出版物显示 NRP2缺陷型小鼠的长时间淋巴水肿，持续肿胀和淋巴引流不足在急性炎症反应期间，该提案为该提案提供了有力的前提。我们的新老鼠在淋巴内皮中缺乏NRP2的淋巴管生成。另外，新数据证明FOXC2负调控NRP2（以及VEGFR3）的表达。在一起，这些数据表明，NRP2对于正常分辨出分液的液体是必需的。我们假设可以操纵NRP2信号轴，以在淋巴功能障碍和慢性炎症。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。 AIM 1将探索成人淋巴管生成中NRP2和VEGFR3信号的基础机制使用组织特异性的遗传消融策略和近红外成像的体内进行评估在炎症的情况下，淋巴引流。鉴于FOXC2下调了NRP2，但NRP2是通常在病理状况下上调，我们将评估NRP2介导的增强成人野生型和淋巴特异性FOXC2功能丧失小鼠的淋巴功能。 AIM 2将检查调节淋巴内皮中NRP2表达的分子机制。相应地，淋巴特异性FOXC2功能获得的小鼠表现出降低的淋巴功能特定的NRP2缺陷小鼠。如何通过FOXC2调节NRP2表达的理解很少，但是新的数据表明它可能会干扰政变 - TFII活动。 AIM 3将研究治疗潜力抑制内源性SEMA3F治疗淋巴功能障碍。总的来说，这个项目是创新的将产生新型的转基因动物模型，揭示淋巴的不同调节分子机制内皮，并提供原始的潜在治疗靶标。总而言之，我们的发现可以翻译进入包括淋巴水肿在内的淋巴功能障碍的创新方法。