Kidney Microphysiological Analysis Platforms (MAP) to Optimize Function and Model Disease

用于优化功能和疾病模型的肾脏微生理分析平台 (MAP)

基本信息

批准号：
10018126
负责人：
JOSEPH VINCENT BONVENTRE
金额：
$ 100.61万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-25 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10018126
关键词：
3-Dimensional 3D Print Acute Acute Disease Adult Affect Alleles Apoptosis Architecture Basement membrane Biocompatible Materials Biological Biology Blood Vessels CRISPR/Cas technology Cell Communication Cell Cycle Cells Cellular Structures Characteristics Chronic Disease Chronic Kidney Failure Clinical Sciences Clustered Regularly Interspaced Short Palindromic Repeats Cyst Cystic Kidney Diseases Cystic kidney Cytoskeleton Development Devices Disease Disease model Distal Drug toxicity Electroencephalography Endothelial Cells Endothelium Engineering Epithelial Epithelial Cells Epithelial Physiology Epithelium Extracellular Matrix Genetic Genetic Diseases Genetic Models Growth Factor Human Image In Vitro Individual Induced Mutation Injury Kidney Kidney Diseases Laboratories Mendelian disorder Microfabrication Microfluidics Modeling Monitor Mutation Nephrons Organoids Patients Perfusion Pericytes Pharmaceutical Preparations Physiologic Monitoring Physiological Population Prevalence Preventive Process Property Protocols documentation Renal glomerular disease Reporter Reproducibility Research Personnel Side Signal Transduction Structure System Techniques Testing Therapeutic Therapeutic Agents Time Tissues Toxic effect Toxicity Tests Tubular formation Work bioprinting cost design design and construction disease phenotype drug development drug discovery drug efficacy drug testing efficacy testing experience genome editing glomerular basement membrane human pluripotent stem cell human tissue induced pluripotent stem cell innovation kidney cell kidney vascular structure microphysiology system mutant nephrotoxicity new technology non-genetic novel organ on a chip planar cell polarity podocyte prevent programs response screening stem cell biology stem cells uptake vector

项目摘要

Project Summary Approximately 10% of the world's adult population has chronic kidney disease (CKD) for which there are very few effective preventive or stabilizing therapeutic options. In addition, 30% of newly developed drugs are not advanced because of nephrotoxicity. We have developed efficient directed differentiation protocols to generate nephron progenitor cells (NPCs) and 3D kidney organoids from human pluripotent stem cells (hPSCs). At present, however, there are no effective platforms that integrate these kidney cells and vascularized organoids within microphysiological systems in vitro to develop effective kidney models for interrogation of nephrotoxicity and drug efficacy. Our proposal unites expertise in kidney organoids and disease, microphysiological systems, and bioprinting led by three experienced investigators (Bonventre, Lee and Lewis) in a unique effort to create these needed model platforms. In Specific Aim 1 we will develop efficient processes to direct differentiation of hiPSCs into kidney podocytes, tubular epithelial cells, and endothelial cells endowed with differentiated features for integration into microphysiological analysis platforms (MAP) and bioprinted structures. We create genetic models of disease and reporter lines that signal differentiation characteristics to optimize differentiation protocols and to monitor physiological parameters. In Specific Aim 2 we will design, construct, and characterize an integrated kidney MAP to evaluate the function of hPSC-derived kidney podocytes, endothelial and epithelial cells as well as kidney organoids. We will also use this platform to create a model of a glomerulus that will have differentiated podocytes on an extracellular matrix (to mimic the glomerular basement membrane) and hiPSC-derived endothelial cells on the other side of the basement membrane. The MAP will be optimized to interrogate basic kidney biology and pathobiology of both non-genetic and genetic disease involving kidney cysts or podocyte injury and test responses to putative therapeutic agents. In Specific Aim 3 we will bioprint a 3D kidney model that contains convoluted proximal tubules, pericytes and endothelial-lined vascular structures with controlled, physiologically relevant system. Modeled tubules and vasculature will be perfused through a open lumens. The ECM composition will be optimized to support confluent epithelialization using proximal and distal tubule cells, podocytes, and endothelial cells derived from hiPSCs. We will characterize polarized drug uptake, toxicity, and vectorial transport through the interstitium (ECM) as well as cell-cell interactions among the epithelial cells, interstitium and endothelium-lined channels to create and validate vascularized kidney models composed of cells derived from healthy and patients with cystic disease that affects the tubule and glomerular disease that affects the podocyte. Our program, with well-established milestones, will result in novel models to test kidney toxicity and drug efficacy.

项目概要世界上大约 10% 的成年人患有慢性肾病 (CKD)，对此有很多治疗方法。有效的预防或稳定治疗选择很少。此外，新开发的药物中有30%不是由于肾毒性而进展。我们开发了高效的定向分化方案来生成来自人类多能干细胞 (hPSC) 的肾单位祖细胞 (NPC) 和 3D 肾脏类器官。在然而，目前还没有有效的平台将这些肾细胞和血管化类器官整合起来在体外微生理系统内开发有效的肾脏模型来研究肾毒性和药物疗效。我们的提案结合了肾脏类器官和疾病、微生理系统、和生物打印由三位经验丰富的研究人员（Bonventre、Lee 和 Lewis）领导，以独特的努力创造这些需要的模型平台。在具体目标 1 中，我们将开发有效的流程来直接区分 hiPSC 分化为肾足细胞、肾小管上皮细胞和内皮细胞集成到微生理分析平台（MAP）和生物打印结构中的功能。我们创造疾病遗传模型和报告细胞系，发出分化特征信号以优化分化协议并监测生理参数。在具体目标 2 中，我们将设计、建造和表征集成的肾脏 MAP，以评估 hPSC 衍生的肾足细胞、内皮细胞的功能和上皮细胞以及肾脏类器官。我们还将利用这个平台来创建一个模型肾小球在细胞外基质上具有分化的足细胞（以模拟肾小球基底）膜）和 hiPSC 衍生的内皮细胞位于基底膜的另一侧。地图将进行优化以询问非遗传性和遗传性疾病的基础肾脏生物学和病理学涉及肾囊肿或足细胞损伤并测试对假定治疗药物的反应。具体目标 3 我们将生物打印一个 3D 肾脏模型，其中包含近曲小管、周细胞和内皮细胞具有受控的、生理相关系统的血管结构。模型化的肾小管和脉管系统将是通过开放的管腔进行灌注。 ECM 成分将被优化以支持汇合上皮化使用源自 hiPSC 的近端和远端肾小管细胞、足细胞和内皮细胞。我们将表征极化药物摄取、毒性和通过间质 (ECM) 的矢量转运以及上皮细胞、间质和内皮细胞之间的细胞间相互作用，以产生和验证由健康细胞和囊性疾病患者细胞组成的血管化肾脏模型影响肾小管和肾小球疾病，影响足细胞。我们的计划，具有完善的里程碑，将产生测试肾毒性和药物疗效的新模型。