Engineering a Physiomimetic Glomerulus-on-Chip to Model Diabetic Kidney Disease

设计拟态肾小球芯片来模拟糖尿病肾病

• 批准号: 9767521
• 负责人: Matthew Mohamed Ishahak
• 金额: $ 3.08万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2020-05-06
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9767521
• 关键词: 3-Dimensional; Address; Adopted; Affect; Albumins; Animal Model; Area; Biochemical; Biological; Biological Assay; Biomedical Engineering; Biomimetics; Blood Pressure; Blood capillaries; Cell Communication; Cell Culture Techniques; Cell Line; Cells; Chemicals; Clinical; Clinical Research; Complex; Computer Assisted; Custom; Development; Device Designs; Devices; Diabetes Mellitus; Diabetic Nephropathy; Disease model; Endothelial Cells; Endothelium; Engineering; Engravings; Epithelial Cells; Equipment; Exposure to; Extracellular Matrix; FDA approved; Filtration; Functional disorder; Gene Expression; Gene Proteins; Glucose; Goals; Human; Hydrogels; In Vitro; Injury; Inulin; Kidney; Kidney Diseases; Kidney Glomerulus; Label; Lasers; Liquid substance; Measures; Methodology; Microfluidics; Modeling; Nephrology; Nephrons; Output; Pathogenesis; Pathology; Patients; Perfusion; Pharmaceutical Preparations; Pharmacologic Substance; Phenotype; Physiological; Physiology; Plastics; Prevalence; Proteins; Research; Serum; Side; Signal Transduction; Specialized Epithelial Cell; Stress; Structure; System; Techniques; Testing; Therapeutic; Translating; Validation; base; clinically relevant; design; disease phenotype; drug development; drug discovery; effective therapy; engineering design; glomerular basement membrane; glomerular endothelium; glomerular filtration; nephrogenesis; novel; novel therapeutics; organ on a chip; podocyte; pressure; prototype; shear stress; tool; urinary

Project Summary/Abstract Diabetic kidney disease (DKD) affects up to 40% of patients with diabetes. Despite the increasing use of glucose- and blood pressure- lowering medication, the prevalence of DKD is on the rise. Injury and dysfunction of the specialized epithelial cells in the kidney glomeruli have been shown to be a fundamental component of DKD pathology. However, therapeutic discovery for DKD, and kidney disease in general, has lagged other areas due to the lack of assays that faithfully capture the complex pathophysiology of nephropathy. Organs-on-chips are quickly emerging as novel in-vitro platforms to model diseases and test potentially therapeutic compounds. As organs-on-chips are adopted for this purpose, there is a need for in-depth platform characterization and validated disease models. Therefore, the development of a DKD model in a glomerulus-on-chip will serve as a valuable research tool. In this proposal, we seek to engineer a physiomimetic model of the glomerulus, which can be utilized to study the pathophysiology of DKD. To achieve this goal, a thorough engineering methodology will be implemented to design and build an organ-on-chip platform to recapitulate the microenvironment of the glomerular filtration barrier. Based on the output of the engineering design, rapid prototyping equipment will be used to build a PDMS-free organ-on-chip for cell culture and custom fluid handling systems will be implemented to recapitulate physiologic pressures found in the glomerulus. The differentiation and culture of a conditionally immortalized human podocyte cell line through exposure to microenvironmental stresses, such as extracellular matrix stiffness, chemical signals, and pressure gradients, will be optimized on the platform. It is hypothesized that inclusion of these microenvironmental features will enhance the expression of podocyte specific genes and proteins required for maintaining the glomerular filtration barrier. Next, functionality of the engineered glomerulus- on-chip will be assessed via a filtration assay using clinically relevant proteins. Finally, DKD will be induced and characterized on the glomerulus-on-chip platform. Induction of the disease phenotype will be accomplished by exposing cells to sera isolated from patients with DKD. This approach is more advantageous than artificial inducers of injury, which decrease the likelihood that potentially therapeutic compounds will be beneficial in a clinical context. It is hypothesized that exposure to patient sera will result in both phenotypic and functional changes. Changes in filtration function will be assessed by the same filtration assay used to demonstrate normal filtration function. Changes in signaling between glomerular endothelial cells and podocytes will also be assessed. Ultimately, this proposal aims to develop a novel glomerulus-on-chip, which will be able to model the pathophysiology of DKD. Hopefully, this platform will assist in the development of new therapeutic compounds to treat DKD.

项目概要/摘要 高达 40% 的糖尿病患者患有糖尿病肾病 (DKD)。尽管葡萄糖的使用越来越多 和降血压药物的使用，DKD 的患病率正在上升。损伤和功能障碍 肾小球中的特化上皮细胞已被证明是 DKD 的基本组成部分 病理。然而，由于 DKD 和一般肾脏疾病的治疗发现落后于其他领域 缺乏能够忠实捕捉肾病复杂病理生理学的检测方法。器官芯片是 迅速成为新型体外平台，用于模拟疾病和测试潜在的治疗化合物。作为 为此目的采用器官芯片，需要深入的平台表征和验证 疾病模型。因此，在肾小球芯片上开发 DKD 模型将成为有价值的研究成果。 研究工具。在本提案中，我们寻求设计肾小球的拟态模型，该模型可以是 用于研究 DKD 的病理生理学。为了实现这一目标，需要采用彻底的工程方法 实现设计和构建片上器官平台来概括器官的微环境 肾小球滤过屏障。根据工程设计的输出，快速原型设备将被 用于构建用于细胞培养的不含 PDMS 的器官芯片，并将实施定制流体处理系统 概括肾小球中发现的生理压力。有条件的差异化和文化 通过暴露于微环境压力（例如细胞外压力）而永生化的人类足细胞系 基质刚度、化学信号和压力梯度将在该平台上进行优化。据推测 纳入这些微环境特征将增强足细胞特异性基因的表达， 维持肾小球滤过屏障所需的蛋白质。接下来，工程化肾小球的功能- 片上将通过使用临床相关蛋白质的过滤测定进行评估。最后，DKD将被诱导并且 在肾小球芯片平台上进行表征。疾病表型的诱导将通过以下方式完成 将细胞暴露于从 DKD 患者分离的血清中。这种方法比人工方法更有优势 损伤诱导剂，这降低了潜在治疗化合物在疾病中有益的可能性 临床背景。据推测，暴露于患者血清会导致表型和功能性的改变。 变化。过滤功能的变化将通过用于证明正常的相同过滤测定来评估 过滤功能。肾小球内皮细胞和足细胞之间信号传导的变化也会 评估。最终，该提案旨在开发一种新型肾小球芯片，它将能够模拟肾小球 DKD 的病理生理学。希望该平台将有助于开发新的治疗化合物 治疗 DKD。