3D Renal Tissue Chip Models to Evaluate Nephrotoxic Effects of Drugs

用于评估药物肾毒性作用的 3D 肾组织芯片模型

基本信息

批准号：
10249974
负责人：
Leslie Donoghue
金额：
$ 3.73万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10249974
关键词：
3-Dimensional Acute Acute Kidney Tubular Necrosis Address Age Animal Model Animals Architecture Basic Science Biocompatible Materials Biomedical Engineering Blood Vessels Blood capillaries Cell Culture Techniques Cell Line Cell Survival Cell physiology Cells Clinical Trials Coculture Techniques Communities Complex Cues Cultured Cells Dermal Development Devices Diabetic Nephropathy Disease Disease model Drug Evaluation Drug Industry Drug Screening Drug toxicity Drug usage Emerging Technologies Endothelial Cells Engineering Environment Epithelial Cells Evaluation Exhibits Exposure to Failure Fibroblasts Functional disorder Gelatin Generations Glucose Goals Homeostasis Human Hydrogels In Vitro Individual Injury Injury to Kidney Kidney Kidney Diseases Liquid substance Maintenance Measurement Mechanical Stress Mechanics Microfluidics Modeling Nephrons Organ Pathologic Pathway interactions Perfusion Pericytes Pharmaceutical Preparations Pharmacology Phenotype Physiological Play Process Proteins Protocols documentation Proximal Kidney Tubules Race Recreation Renal Tissue Renal function Renal tubular acidosis Research Risk Role Sex Differences Stimulus Stretching Structure System Technology Therapeutic Tight Junctions Tissue Engineering Tissue Microarray Tissues Toxic effect Toxicity Tests Toxin Translational Research Translations Tubular formation Umbilical vein Validation Work age difference base cell injury cell type clinical practice cost cytokine design drug discovery drug efficacy engineering design enzyme activity falls fluid flow glomerular filtration high throughput screening high-throughput drug screening in vitro Model in vivo interstitial model development nephrotoxicity novel organ on a chip patient population pre-clinical preservation pressure prevent prototype racial difference renal tubular dysfunction response screening sex shear stress species difference success three dimensional structure tool toxicant

项目摘要

PROJECT SUMMARY The current pathway for drug discovery is associated with costs of $2.55 billion and between 10-15 years of development for a single drug to reach the market. The challenges in predicting drug toxicities and efficacies are attributed to inherent species differences in drug-metabolizing enzyme activities and cell-type-specific sensitivities to toxicants. Organs-on-a-chip are an emerging technology in disease modeling and screening therapeutics to address discrepancies between animal models and human clinical trials. They utilize tissue engineering, fluid mechanics, and biomaterials to replicate in vivo architectures and functions of complex organs and tissues. The renal proximal tubule (PT) in vivo is exposed to fluid flow and mechanical stress (pressure, stretch, shear) and these stimuli play an important role in maintaining cellular phenotype and homeostasis. Currently, available prototypes fall short of replicating the in vivo environment because they often fail to mimic the physiological forces. Therefore, these models have had limited success in predicting drug-induced nephrotoxicity. In this proposal, we will bioengineer and evaluate a dynamic platform of the PT and study the effects of drugs and tubular dysfunction to establish its potential for translational research. Human renal proximal tubule cells (hRPTECs) will be cultured within gelatin methacryloyl (GelMA) hydrogels under physiological shear and pressure. These devices will also incorporate the diversity in the patient population by using hRPTECs from multiple donors to determine the impact of age, sex, and racial differences on nephrotoxicity effects. Drugs will be classified based on their nephrotoxic risk (high, intermediate, and low) and the platform will incorporate automated readouts to reflect cellular function and viability. Together, this will help investigate more accurate pharmacological and pathological responses and to determine the utility of in vitro perfusion models. Secondly, a more complex and novel bioengineered platform will be developed. This design contains a 3D PT tubule and 3D vascular vessels surrounded by pericyte vascular networks. The platform will then be subjected to physiological shear stress and pressure to demonstrate the flow loop can accurately mimic cellular organization, establishment of tight junctions, maintenance of barrier function, and selective transport as seen in vivo. This device composes of a co-culture of hRPTECs, human umbilical vein endothelial cells (hUVECs), and human dermal fibroblasts (hDF) within a GelMA hydrogel to model an environment where both reabsorption and secretion functions are replicated. Lastly, this proposal investigates the translational potential of PT tissue chips through demonstration of a PT diabetic nephropathy model and engineering multi-well PTs to facilitate high- throughput studies. The organ-on-a-chip developed in this study will provide an enabling technology that has broad applications in basic and translational research to model disease states, study interactions with other tissue chips, and accurately predict drug toxicity.

项目概要目前的药物发现途径需要 25.5 亿美元的成本和 10 至 15 年的时间开发单一药物以进入市场。预测药物毒性和功效的挑战是归因于药物代谢酶活性和细胞类型特异性的固有物种差异对毒物的敏感性。器官芯片是疾病建模和筛查的新兴技术解决动物模型和人体临床试验之间差异的治疗方法。他们利用组织工程、流体力学和生物材料来复制复杂器官的体内结构和功能和纸巾。体内肾近曲小管 (PT) 暴露于液体流动和机械应力（压力、拉伸、剪切），这些刺激在维持细胞表型和稳态方面发挥着重要作用。目前，可用的原型无法复制体内环境，因为它们通常无法模仿生理力量。因此，这些模型在预测药物诱导的肾毒性。在本提案中，我们将对 PT 的动态平台进行生物工程和评估，并研究药物和肾小管功能障碍的影响，以确定其转化研究的潜力。人肾近端肾小管细胞 (hRPTEC) 将在生理剪切力下在明胶甲基丙烯酰 (GelMA) 水凝胶中培养和压力。这些设备还将通过使用 hRPTEC 来整合患者群体的多样性多个捐赠者以确定年龄、性别和种族差异对肾毒性影响的影响。药物会根据肾毒性风险（高、中、低）进行分类，该平台将纳入自动读数反映细胞功能和活力。总之，这将有助于更准确地调查药理学和病理学反应并确定体外灌注模型的效用。第二，将开发更复杂和新颖的生物工程平台。该设计包含 3D PT 小管和被周细胞血管网络包围的 3D 血管。随后该平台将受到生理剪切应力和压力，以证明流动回路可以准确地模拟细胞组织，建立紧密连接、维持屏障功能以及体内选择性运输。这该装置由 hRPTEC、人脐静脉内皮细胞 (hUVEC) 和人脐静脉内皮细胞 (hUVEC) 的共培养物组成。 GelMA 水凝胶内的真皮成纤维细胞 (hDF) 可以模拟重吸收和分泌功能被复制。最后，该提案研究了 PT 组织芯片的转化潜力通过演示 PT 糖尿病肾病模型并设计多孔 PT 来促进高吞吐量研究。本研究开发的器官芯片将提供一种使能技术在基础和转化研究中广泛应用，以模拟疾病状态、研究与其他疾病的相互作用组织芯片，准确预测药物毒性。