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，我们必须确定更好的治疗策略，以避免 医疗危机。目前的治疗干预措施解决患者的症状，而不是根本原因， 因此基本上不会减缓疾病的发展。开发药物来治疗根本原因或 导致CKD的疾病将满足这一需求，然而，这种进展受到缺乏 用于药物开发管道的体外人类肾脏模型。因此，准确地 模拟个体人类CKD，并可筛选疾病相关的治疗干预措施， 需要确定有针对性的CKD干预措施。第二阶段SBIR是第一阶段成功的延续， MIMETAS和洛杉矶儿童医院（CHLA）之间的合作努力。我们将共同 专注于解决市场对3D肾脏芯片平台的需求，称为用于药物的NephroPlate 的发现NephroPlate将能够评价肾功能对循环和遗传因素的反应。 导致CKD的疾病因素。对于这个II期项目，我们将重点关注肾小球疾病， 以往的经验，Alport综合征和膜性肾病，以及哪里有高需求， 糖尿病肾病和APOL1肾病的患者。对于每个CKD，我们将描述 表型（免疫染色，蛋白质印迹，RNAseq）和肾小球滤过功能，通过测量传代 的荧光白蛋白和跨上皮电阻（TEER）的微流体系统内。所有 将根据高通量药物筛选标准对系统进行验证。肾小球芯片的滤液 将培养物转移到近端小管芯片培养物中，以评价肾小球疾病的影响 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分析，以确定 对每种CKD特异性的潜在生物标志物。一旦执行，这些研究将确立NEPHROPLATE作为 高通量药物筛选平台，供制药公司和学术实验室使用， 研究CKD疾病的机制，并最终确定疾病特异性治疗方法。

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.