Modeling atypical hemolytic uremic syndrome with hydrogel-based microvasculature-on-chip technologies

利用基于水凝胶的微血管芯片技术模拟非典型溶血性尿毒症综合征

• 批准号: 9809757
• 负责人: Satheesh Chonat
• 金额: $ 20.95万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9809757
• 关键词: Acute; Affect; Alternative Complement Pathway; Anemia; Animal Model; Animals; Ascorbic Acid; Biological Markers; Biomedical Engineering; Blood; Blood Platelets; Blood Vessels; Blood coagulation; Body part; Cessation of life; Chronic; Chronic Kidney Failure; Clinical; Coagulation Process; Complement; Complement 3a; Complement 5a; Complement Activation; Complement Inactivators; Complement Membrane Attack Complex; Complex; Data; Deposition; Development; Disease; Disease Outcome; End stage renal failure; Endothelium; Engineering; Environment; Erythrocytes; Exhibits; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; Extracellular Matrix; Fibrin; Flare; Fright; Functional disorder; Hematology; Heme; Hemolytic Anemia; Hemolytic-Uremic Syndrome; Hemostatic function; Human; Hydrogels; Immune system; In Vitro; Injury; Kidney; Knowledge; Lead; Life; Malignant Neoplasms; Mediating; Methodology; Microscopic; Modeling; Molecular; Monitor; Monoclonal Antibodies; Natural History; Nature; Nitric Oxide; Outcome; Pathogenesis; Pathway interactions; Patients; Permeability; Pharmaceutical Preparations; Pharmacology; Physiological; Plasma; Plasma Proteins; Platelet Count measurement; Play; Process; Production; Proteins; Reactive Oxygen Species; Recovery; Research; Resolution; Role; Shiga Toxin; Sickle Cell Anemia; System; Technology; Testing; Thrombocytopenia; Thrombosis; Thrombus; Time; Translating; Transplantation; Triad Acrylic Resin; Validation; Vascular Diseases; Work; base; biophysical properties; complement pathway; complement system; drug discovery; endothelial dysfunction; experimental study; improved; in vitro Model; in vivo; in vivo Model; injured; insight; membrane assembly; mortality; novel; novel therapeutics; rare genetic disorder; real time monitoring; repaired; response; shear stress; tool; treatment strategy

PROJECT SUMMARY ABSTRACT Atypical hemolytic uremic syndrome (aHUS) is a life-threatening disease that causes microvascular thrombosis, especially in the kidneys, with a global mortality rate of 25%, and a frequent progression to end-stage renal disease (ESRD)1,2. While aHUS is known to be associated with uncontrolled activation of the alternative pathway (AP) of complement, the underlying pathophysiology of how AP activation causes endothelial injury and thrombosis remains unclear, which largely limits the development of additional therapies for aHUS. Even with the currently most effective drug, eculizumab, many still suffer from acute flare ups, progression to ESRD, and extra-renal manifestations, which are thought to be associated with unregulated endothelial activation. The proposed work aims to develop the first in vitro model of aHUS that incorporates all of the major components thereof, including complement and other plasma proteins, platelets, red blood cells, intact endothelium, extracellular matrix with physiologic biophysical properties, and shear stress, to not only improve our understanding of the pathophysiology of this feared disease but also to explore new therapeutics that may improve better clinical outcomes for aHUS patients. Here we hypothesize that a novel hydrogel-based “endothelialized” microvasculature-on-a-chip that we recently invented recapitulates the in vivo microvascular microenvironment and exhibits long-term (>2 months) microvasculature functionality. This can be used as an in vitro model for aHUS, as our microvasculature-on-a-chip system enables the decoupling of the complex cellular and molecular factors (including complement and regulators of complement activation) involved in the pathophysiology of aHUS as well as quantitatively characterizes how these factors interact and lead to endothelial injury or activation. These factors will be perfused into the engineered microvasculature, and biomarkers of endothelial dysfunction, such as endothelial permeability, reduced nitric oxide and increased reactive oxygen species, will be monitored in real time. In addition, the system will allow assessment of how these factors synergistically lead to thrombi formation in the engineered microvasculature in the context of aHUS through quantifying fibrin formation and platelet aggregate/microthrombi size. As eculizumab only blocks the formation of the membrane attack complex in the terminal pathway of complement activation, this microvasculature-on-a-chip model will enable the testing of other novel treatment strategies to promote endothelial repair. Successful completion of this study will result in the development of a novel and robust model for aHUS, gain a better understanding of endothelial damage in aHUS and more broadly how the complement system interacts with hemostasis and thrombosis, and provide insights into developing rational pharmacological approaches in the management of aHUS and other thrombotic microangiopathies.

项目摘要摘要 非典型溶血性尿毒症综合征（AHU）是一种威胁生命的疾病，会导致微血管血栓形成， 尤其是在儿童中，全球死亡率为25％，并且频繁发展为终端肾脏 疾病（ESRD）1,2。虽然已知AHU与替代途径的不受控制激活有关 （AP）完成，AP激活如何导致内皮损伤和 血栓形成尚不清楚，这在很大程度上限制了针对AHU的其他疗法的发展。即使有 目前最有效的药物，eculizumab，许多人仍然患有急性爆发，进展到ESRD，并且 肾外表现，被认为与不受监管的内皮激活有关。 拟议的工作旨在开发首个结合所有专业的体外模型 其成分，包括完成和其他血浆蛋白，血小板，红细胞，完整 内皮，具有物理生物物理特性的细胞外基质和剪切应力，不仅 提高我们对这种Feeed疾病的病理生理学的理解，但也探索新疗法 这可能会改善艾哈斯患者的更好临床结果。在这里，我们假设一种新型水凝胶 我们最近发明的“内皮化”片上的微脉管系统概括了体内微血管 微环境并表现出长期（> 2个月）的微脉管功能。这可以用作 Ahus的体外模型，因为我们在芯片上的微脉管系统可以使复杂细胞的脱钩 和分子因素（包括完整激活的补体和调节剂） Ahus的病理生理以及定量表征这些因素如何相互作用并导致 内皮损伤或激活。这些因素将被灌注到工程的微脉管系统中，并且 内皮功能障碍的生物标志物，例如内皮通透性，一氧化氮降低并增加 活性氧将进行实时监测。此外，该系统将允许评估如何 这些因素在Ahus的背景下协同导致工程微脉管系统中的血栓形成 通过量化纤维蛋白形成和血小板骨料/微骨大小。由于eculizumab只能阻止 膜攻击复合物在完成终端途径中的形成，这是 片上的微脉管模型将使其他新型治疗策略能够促进 内皮修复。 这项研究的成功完成将导致开发一个新颖而健壮的痤疮模型，获得 更好地理解AHUS中的内皮损害以及更广泛的完成系统如何相互作用 止血和血栓形成，并提供有关开发理性药物方法的见解 Ahus和其他血栓形成微动病的管理。