Development of the NephroPlate: A high-throughput kidney-on-a-chip platform for identifying chronic kidney disease therapies

项目摘要 随着慢性肾脏疾病的增加，我们必须找到更好的治疗策略，以避免 医疗保健危机。目前的治疗干预措施针对的是患者的症状，而不是不充分的原因和 因此不会实质上减缓疾病的进展。开发治疗根本原因的药物或 导致CKD的紊乱将解决这一需求，然而，这种进步因缺乏而受到阻碍 用于药物开发流水线的体外人类肾脏模型。因此，一种准确的体外模型 模拟单个人类慢性KD并可筛查与疾病相关的治疗干预措施迫在眉睫 需要确定有针对性的慢性肾脏病干预措施。这一第二阶段SBIR是成功的第一阶段和 MIMETAS和洛杉矶儿童医院(CHLA)的合作努力。我们将携手共进 专注于满足市场对芯片上3D肾脏平台的需求，该平台被称为用于药物的肾板 发现号。肾板将能够评估肾功能对循环和遗传的反应 慢性肾脏病的致病因素。对于这个第二阶段的项目，我们将专注于我们有 既往经验，阿尔波特综合征和膜性肾病，以及对 肾板客户、糖尿病肾病和APOL1肾病。对于每个CKD，我们将描述 表型(免疫染色、免疫印迹、RNAseq)和肾小球滤过功能的测定 荧光白蛋白和微流控系统中的跨上皮电阻值(TEER)。全 系统将按照高通量药物筛选标准进行验证。芯片上肾小球的滤液 培养物将被转移到近端芯片上的小管培养物以评估肾小球疾病的影响。 在近端小管上。最后，将对肾小球和近端小管滤液进行蛋白质组学研究，以确定 每种CKD的潜在生物标记物。一旦执行，这些研究将建立起肾盘作为 供制药公司和学术实验室使用的高通量药物筛选平台 研究慢性肾脏病的发病机制，并最终确定针对疾病的治疗方法。

Project Summary With chronic kidney disease on the rise, it is imperative that we identify better treatment strategies to avoid a healthcare crisis. Current therapeutic interventions address patient symptoms rather than the underly cause and thereby do not substantially slow disease advancement. Developing drugs to treat the underlying cause or disorder leading to CKD will address this need, however such advancements have been hampered by the lack of in vitro human kidney models used in the drug development pipeline. Thus, an in vitro model which accurately mimics individual human CKDs and can be screened for disease-relevant therapeutic interventions is urgently needed to identify targeted CKD interventions. This Phase II SBIR is a continuation of a successful Phase I and a collaborative effort between MIMETAS and the Children’s Hospital of Los Angeles (CHLA). Together we will focus on addressing the market need for a 3D kidney-on-a-chip platform, called the NephroPlate for use in drug discovery. The NephroPlate will enable the evaluation of kidney function in response to circulating and genetic factors of CKD-causing disorders. For this Phase II project, we will focus on glomerulopathies where we have previous experience, Alport syndrome and membranous nephropathy, and where there is a high demand from NephroPlate customers, diabetic nephropathy and APOL1 nephropathy. For each CKD, we will characterize phenotype (immunostaining, western blot, RNAseq) and glomerular filtration function by measuring the passage of fluorescent albumin and the trans-epithelial electrical resistance (TEER) within the microfluidic system. All systems will be validated to high-throughput drug screening standards. Filtrate from the glomerulus-on-a-chip cultures will be transferred to the proximal tubule-on-a-chip cultures to evaluate the impact of glomerular disease on proximal tubules. Finally, proteomics will be performed on the glomerular and proximal tubule filtrate to identify potential biomarkers specific to each CKD. Once executed these studies will establish the NephroPlate as a high-throughput drug screening platform for use by pharmaceutical companies and academic laboratories to investigate mechanisms of CKD diseases and ultimately identify disease-specific therapeutics.