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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要摘要非典型溶血性尿毒症综合征（aHUS）是一种危及生命的疾病，会导致微血管血栓形成，尤其是肾脏，全球死亡率为 25%，并且经常进展为终末期肾病疾病 (ESRD)1,2，而已知 aHUS 与旁路途径的不受控制的激活有关。补体 (AP)，AP 激活如何导致内皮损伤的潜在病理生理学血栓形成仍不清楚，这在很大程度上限制了 aHUS 额外疗法的开发。目前最有效的药物依库丽单抗（eculizumab），许多人仍然遭受急性发作、进展为 ESRD 的困扰，并且肾外表现，被认为与不受调节的内皮激活有关。拟议的工作旨在开发第一个 aHUS 体外模型，其中包含所有主要功能完整的成分，包括补体和其他血浆蛋白、血小板、红细胞内皮、具有生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可怕疾病的病理生理学的理解，同时探索新的治疗方法这可能会改善 aHUS 患者更好的临床结果。在这里，我们致力于开发一种新型的基于水凝胶的药物。我们最近发明的“内皮化”微血管芯片概括了体内微血管微环境并表现出长期（> 2 个月）微血管功能，这可以用作内部。 aHUS 的体外模型，因为我们的微血管芯片系统能够解耦复杂的细胞和参与的分子因素（包括补体和补体激活的调节因子） aHUS 的病理生理学以及定量描述这些因素如何相互作用并导致这些因子将被灌注到工程化的微血管系统中，并且内皮功能障碍的生物标志物，例如内皮通透性、一氧化氮减少和增加此外，系统还可以评估活性氧的情况。这些因素协同导致 aHUS 背景下工程化微血管系统中的血栓形成通过量化纤维蛋白形成和血小板聚集/微血栓大小，因为依库丽单抗仅阻断在补体激活的终末途径中形成膜攻击复合物，这芯片上的微血管模型将能够测试其他新颖的治疗策略，以促进内皮修复。这项研究的成功完成将为 aHUS 开发一种新颖且稳健的模型，获得更好地了解 aHUS 中的内皮损伤以及更广泛的补体系统如何相互作用止血和血栓形成，并为开发合理的药理学方法提供见解 aHUS 和其他血栓性微血管病的治疗。