Regulation of Lymphatic Endothelial Cell Junction and Drainage

淋巴内皮细胞连接和引流的调节

• 批准号: 10642883
• 负责人: Esak Lee
• 金额: $ 47.54万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10642883
• 关键词: 3-Dimensional; Alzheimer' s Disease; Animal Model; Architecture; Arthritis; Binding; Biological; Biology; Biomedical Engineering; Biophysics; Blood Vessels; Boston; Brain; Cell Culture Techniques; Cells; Clinical Research; Collaborations; Complex; Development; Dietary Fats; Disease; Drainage procedure; Exhibits; Experimental Models; Fibronectins; Fibrosis; Fluid Balance; Glaucoma; Goals; Homeostasis; Human; ITGA5 gene; Immune System Diseases; Immunity; Immunologist; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Intercellular Fluid; Intercellular Junctions; Ligands; Liquid substance; Lymph; Lymphatic; Lymphatic Diseases; Lymphatic Endothelial Cells; Lymphatic Endothelium; Lymphatic System; Lymphatic function; Lymphedema; Malignant Neoplasms; Mediating; Meningeal lymphatic system; Metabolic Diseases; Modeling; Morphogenesis; Morphology; Mus; Neurodegenerative Disorders; Neurons; Obesity; Organ; Outcome; Pediatric Hospitals; Pericytes; Permeability; Physiological; Protein Kinase; ROCK1 gene; Regulation; Research Personnel; Role; Signal Transduction; Small Intestines; Structure; Tissues; Treatment Efficacy; Universities; Vascular System; drug candidate; human disease; in vitro Model; in vivo; in vivo Model; inhibitor; lacteal; lymph nodes; lymphatic drainage; lymphatic dysfunction; lymphatic vessel; rho; screening; therapeutic evaluation; three-dimensional modeling; tool; two-dimensional; uptake; wasting

PROJECT SUMMARY Lymphatic vessel (LV) differentiation, development, and morphogenesis are central in maintaining fluid homeostasis, regulating host immunity, and transporting dietary fat and neuronal waste. All these functions are governed by lymphatic drainage, a transport of interstitial fluid into the lymphatic system through the initial LVs and collecting LVs. The initial LVs show permeable button-like junction morphology and are ready to uptake interstitial fluid; by contrast, the collecting LVs are less permeable with zipper-like junction structure, so that the collecting LVs transport ‘lymph’ to lymph nodes without leaking. Impaired lymphatic drainage contributes to many human diseases, such as lymphedema, immune dysfunction, fibrosis, obesity, cancer, and Alzheimer’s disease. While little is known about why LVs become dysfunctional, clinical studies reveal that inflammation is one of the leading contributors to the lymphatic dysfunction. Although dysfunctional collecting LVs has been extensively studied, how inflammation impacts initial LV development and morphogenesis is unclear, because in our current experimental models, including animal models, we often cannot decouple multifactorial inflammatory factors in the lymphatic endothelium. Since two-dimensional cell culture has failed to recapitulate three-dimensional (3D) tissue architecture of lymphatics, researchers have developed 3D in vitro models of LVs, demonstrating lymphatic sprouting, lymphatic network formation, and LV interactions with other cells. However, these previous models have not created 3D lymphatic structure with specialized LEC junction development enabling controlled fluid drainage through the button-like junctions and physiological inflammatory response. In this proposal, we will use a bioengineered in vitro 3D lymphatic vascular system, exhibiting button-like junction morphogenesis of the LVs and fluid drainage to understand the regulation of LEC junction and drainage by focusing on ROCK1/2 and integrin α5 signaling. In Aim 1, we will examine the roles of ROCKs in LEC junction and drainage. Next, we will scrutinize the mechanisms of ROCKs-mediated junction zippering in LECs. In Aim 2, we will study integrin α5 mediated regulation of LEC junction and lymphatic drainage. We will then determine signal transduction through ROCKs and integrin α5 and evaluate therapeutic efficacy of targeting ROCKs and integrin α5 in lymphatic dysfunction and inflammation models in vivo. In summary, we will use a bioengineered model of 3D lymphatic vessels and fluid transport to provide an understanding of lymphatic drainage in normal and inflammatory conditions.

项目摘要 淋巴管（LV）的分化、发育和形态发生是维持体液平衡的核心。 体内平衡，调节宿主免疫，以及转运膳食脂肪和神经元废物。所有这些功能 是由淋巴引流，间质液进入淋巴系统的运输，通过最初的 LV和收集LV。最初的LV显示出可渗透的纽扣状连接形态，并准备 摄取间质液;相比之下，收集LV的渗透性较低，具有拉链状连接结构， 这样收集的LV将“淋巴液”输送到淋巴结而不会泄漏。淋巴引流受损 导致许多人类疾病，例如水肿、免疫功能障碍、纤维化、肥胖、癌症， 和老年痴呆症尽管人们对左心室功能障碍的原因知之甚少，但临床研究表明， 炎症是导致淋巴功能障碍的主要因素之一。虽然功能失调 收集LV已被广泛研究，炎症如何影响初始LV发育， 形态发生是不清楚的，因为在我们目前的实验模型，包括动物模型，我们经常 不能分离淋巴内皮中的多因子炎症因子。由于二维单元格 培养未能概括三维（3D）组织结构的生物学，研究人员已经 开发了LV的3D体外模型，展示了淋巴发芽，淋巴网络形成， LV与其他细胞的相互作用。然而，这些先前的模型没有创建3D淋巴结构 通过专门的LEC接头开发， 连接和生理炎症反应。在这项提案中，我们将使用生物工程体外3D 淋巴管系统，表现出LV的纽扣样连接形态发生和液体引流， 通过关注ROCK 1/2和整合素α5信号转导，了解LEC连接和引流的调节。 在目标1中，我们将研究ROCKs在LEC连接和引流中的作用。接下来，我们将仔细研究 在LEC中ROCKs介导的连接拉链机制。目的2：研究整合素α5介导的 调节LEC连接和淋巴引流。然后，我们将确定信号转导通过 ROCKs和整合素α 5靶向治疗淋巴结转移的疗效评价 功能障碍和体内炎症模型。总之，我们将使用生物工程模型的3D淋巴 血管和液体运输，以了解正常和炎症中的淋巴引流 条件