A Bioengineered Model for Deciphering Lymphatic Dysfunction in Inflammation

破译炎症中淋巴功能障碍的生物工程模型

基本信息

批准号：
10493273
负责人：
Esak Lee
金额：
$ 21.34万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-22 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10493273
关键词：
3-Dimensional 3D Print Address Affect Alzheimer&apos s Disease Animal Model Architecture Biological Biology Biomedical Engineering Biophysics Blood Vessels Boston Brain Cell Culture Techniques Cells Characteristics Clinical Research Collaborations Dendritic Cells Dermis Dextrans Dietary Fats Diffuse Drainage procedure Ear Experimental Models Extracellular Matrix Fibrosis Fluid Balance Functional disorder Gastrointestinal tract structure Gene Expression Geometry Goals Homeostasis Human Immune Immune System Diseases Immunity Immunologist Impairment In Vitro Inflammation Inflammatory Inflammatory Response Intercellular Fluid Intercellular Junctions Liquid substance Lymph Lymphatic Lymphatic System Lymphatic function Lymphedema Lymphocyte Malignant Neoplasms Measures Meningeal lymphatic system Metabolic Diseases Modeling Mus Neurodegenerative Disorders Neurons Obesity Organ Pathway interactions Pattern Pediatric Hospitals Permeability Pharmaceutical Preparations Physiological Play Process Research Personnel Resolution Role Skin Structure System Thick Tissues Tracer Vascular System cytokine draining lymph node drug candidate human disease in vitro Model in vivo in vivo Model interstitial cell lacteal lymph nodes lymphatic drainage lymphatic dysfunction lymphatic vessel mechanotransduction new therapeutic target screening three-dimensional modeling tool trafficking two-dimensional uptake wasting

项目摘要

PROJECT SUMMARY Lymphatic vessels (LVs) 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 and immune cells into the lymphatic system through initial LVs and collecting LVs. The initial and collecting LVs have different structures and roles in maintaining lymphatic drainage. The initial LVs have permeable cell-cell junctions and are ready to uptake interstitial fluid and cells; by contrast, the collecting LVs are less permeable, so that the collecting LVs can transport ‘lymph’ to lymph nodes without leaking. Impaired lymphatic drainage contributes to LV-related human diseases, such as lymphedema, immune dysfunction, fibrosis, obesity, cancer, Alzheimer’s disease, etc. While little is known about why LVs become dysfunctional, clinical studies reveal that inflammation is one of the leading contributors to the lymphatic dysfunction. Mechanisms of how inflammation affects both initial and collecting LVs making them dysfunctional are unclear, because in our current experimental models, including animal models, we often cannot decouple multifactorial inflammatory factors in the initial and collecting LVs. 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, LV permeability, and LV interactions with surrounding cells. However, these previous models have not created 3D lymphatics with LEC junctions enabling controlled drainage, valve formation, or physiological inflammatory response. Also, they have not considered the distinct roles of initial and collecting LVs. In this proposal, we will establish a bioengineered in vitro 3D lymphatic vascular system, including both initial and collecting LVs, and determine how inflammatory conditions make the initial and collecting LVs dysfunctional. In Aim 1, we will establish collecting LV-on-chip, considering (i) tightened cell-cell junctions, (ii) luminal valves, and (iii) mural cell coverage (Aim 1.1). Next, we will combine the collecting LVs with the initial LVs and demonstrate fluid and immune cell transport through these LVs in the normal condition (Aim 1.2). In Aim 2, we will determine the effects of inflammatory conditions by using inflammatory cytokines that are relevant to lymphatic dysfunction on the initial and collecting LVs and overall lymphatic drainage (Aim 2.1). Next, we will identify new targets to reverse the lymphatic dysfunction by screening pathways of lymphatic vascular mechanotransduction (Aim 2.2). We will then validate the mechanisms and targets in an in vivo skin inflammation model (Aim 2.3). In summary, we will develop a new bioengineered model of 3D lymphatic vascular system and fluid/immune cell transport through the initial and collecting LVs and provide a unique in vitro platform to address how the LV dysfunction occurs in inflammation, and identify new therapeutic targets to reverse the dysfunction.

项目概要淋巴管 (LV) 在维持体液稳态、调节宿主免疫力和运输膳食脂肪和神经废物所有这些功能都由淋巴引流控制。通过初始左室和集合左室将间质液和免疫细胞转运至淋巴系统。初始左心室和集合左心室在维持淋巴引流方面具有不同的结构和作用。相比之下，左心室具有可渗透的细胞-细胞连接，并准备好吸收间质液和细胞；收集左室的渗透性较差，因此收集左室可以将“淋巴”运输到淋巴结，而无需淋巴引流受损会导致与左心室相关的人类疾病，例如淋巴水肿、免疫功能障碍、纤维化、肥胖、癌症、阿尔茨海默氏病等。虽然人们对 LV 的原因知之甚少变得功能失调，临床研究表明炎症是导致功能失调的主要原因之一炎症如何影响初始左心室和集合左心室的机制。功能失调尚不清楚，因为在我们目前的实验模型，包括动物模型中，我们经常由于二维，无法解耦初始 LV 和收集 LV 中的多因素炎症因子。研究人员表示，细胞培养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，左室通透性以及左室与周围细胞的相互作用然而，这些先前的模型没有。创建具有 LEC 连接的 3D 淋巴管，可实现受控引流、瓣膜形成或生理学此外，他们没有考虑初始 LV 和收集 LV 的不同作用。根据提案，我们将建立一个生物工程体外3D淋巴血管系统，包括初始和收集左室，并确定炎症条件如何使初始左室和收集左室功能障碍。在目标 1 中，我们将建立收集 LV 芯片，考虑 (i) 紧密的细胞与细胞连接，(ii) 腔阀， (iii) 壁细胞覆盖（目标 1.1）接下来，我们将收集 LV 与初始 LV 结合起来。证明正常情况下液体和免疫细胞通过这些 LV 运输（目标 1.2）。我们将通过使用与以下相关的炎症细胞因子来确定炎症状况的影响初始左心室和集合左心室以及整体淋巴引流的淋巴功能障碍（目标 2.1）。通过筛选淋巴血管通路，确定逆转淋巴功能障碍的新靶点然后，我们将验证体内皮肤的机制和目标。炎症模型（目标 2.3）总之，我们将开发一种新的 3D 淋巴管生物工程模型。血管系统和液体/免疫细胞通过初始和集合 LV 运输，并提供独特的体外平台来解决炎症中左心室功能障碍如何发生，并确定新的治疗方法目标是逆转功能障碍。