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: 10502991
• 负责人: Esak Lee
• 金额: $ 48.34万
• 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/10502991
• 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; 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; Play; 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.

项目概要 淋巴管 (LV) 分化、发育和形态发生对于维持液体至关重要 体内平衡，调节宿主免疫力，以及运输膳食脂肪和神经元废物。所有这些功能 受淋巴引流控制，即间质液通过最初的淋巴管输送到淋巴系统 LV 和收藏 LV。最初的 LV 显示出可渗透的纽扣状连接形态，并准备好 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， 以便收集左心室将“淋巴”输送到淋巴结而不发生泄漏。淋巴引流受损 导致许多人类疾病，如淋巴水肿、免疫功能障碍、纤维化、肥胖、癌症、 和阿尔茨海默病。虽然我们对左心室功能失调的原因知之甚少，但临床研究表明 炎症是淋巴功能障碍的主要原因之一。虽然功能失调 收集 LV 已被广泛研究，炎症如何影响 LV 的初始发育和 形态发生尚不清楚，因为在我们目前的实验模型，包括动物模型中，我们经常 不能解耦淋巴管内皮中的多因素炎症因子。由于二维细胞 研究人员发现，培养物未能重现淋巴管的三维 (3D) 组织结构 开发了 LV 的 3D 体外模型，展示了淋巴萌芽、淋巴网络形成，以及 左心室与其他细胞的相互作用。然而，之前的这些模型并没有创建 3D 淋巴结构 具有专门的 LEC 连接开发，可通过按钮状控制液体排出 连接和生理炎症反应。在本提案中，我们将使用生物工程体外 3D 淋巴管系统，表现出左心室的纽扣状连接形态发生和液体引流 通过关注 ROCK1/2 和整合素 α5 信号传导来了解 LEC 连接和引流的调节。 在目标 1 中，我们将研究 ROCK 在 LEC 连接和引流中的作用。接下来，我们将仔细审视 ROCKs介导的LECs连接拉链机制。在目标 2 中，我们将研究整合素 α5 介导的 LEC 连接和淋巴引流的调节。然后我们将通过以下方式确定信号转导 ROCKs 和整合素 α5 并评估淋巴管中靶向 ROCKs 和整合素 α5 的治疗效果 体内功能障碍和炎症模型。总之，我们将使用 3D 淋巴管的生物工程模型 血管和液体运输，以了解正常和炎症的淋巴引流 状况。