High-throughput Flow Culture of 3D Human PKD Models for Therapeutic Screening

用于治疗筛选的 3D 人体 PKD 模型的高通量流式培养

基本信息

批准号：
10649222
负责人：
Neil Lin
金额：
$ 23.5万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-15 至 2025-05-14
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10649222
关键词：
Acceleration Adult Affect Algorithms Animal Model Apoptosis Arrhythmia Artificial Intelligence Autosomal Dominant Polycystic Kidney CRISPR/Cas technology Calcium Cardiovascular system Cell model Cells Cessation of life Chronic Kidney Failure Conceptions Coronary heart disease Cyst Cystic Fibrosis Transmembrane Conductance Regulator Data Analyses Development Devices Dialysis procedure Diameter Disease Disease model Dropout Duct (organ) structure Ductal Epithelial Cell End stage renal failure Epithelium Exposure to FDA approved FRAP1 gene Fibrosis Generations Genes Genetic Genetic Engineering Genotype Heart failure Heterozygote Homeostasis Human Hypertension Image Immunofluorescence Immunologic Inherited Intervention Kidney Kidney Failure Kidney Transplantation Liquid substance Mediating MicroRNAs Modality Modeling Molecular Morbidity - disease rate Mus Mutation Nephrons Organ Organoids Oxygen PKD2 protein Patients Persons Phenotype Physiological Proliferating Proteins Renal Tissue Renal function Renal tubule structure Renin-Angiotensin-Aldosterone System Risk Source System Technology Testing Therapeutic Tissue Model United States Urine Vasopressin Receptor analysis pipeline antagonist automated analysis body system cell type data analysis pipeline density design drug candidate drug development efficacious treatment fluid flow fluorescence imaging high throughput screening improved in vivo indexing induced pluripotent stem cell inhibitor innovation interstitial kidney cell kidney epithelial cell malformation novel novel therapeutics polycystic kidney disease 1 protein response screening single nucleus RNA-sequencing standard of care symptom management therapeutic development three dimensional cell culture three dimensional structure tissue culture tolvaptan urinary

项目摘要

Autosomal Dominant Polycystic Kidney Disease (ADPKD) is a genetically inherited, multi-organ-system disease that is characterized by the formation of fluid filled cysts and intertubular fibrosis in the kidney. This results in a gradual decline in kidney function ultimately leading to kidney failure. Importantly, cardiovascular complications are the main cause of morbidity and death in ADPKD patients. This is a direct result of the over- activation of renin-angiotensin-aldosterone system, which causes hypertension and an increased risk for coronary heart disease, arrhythmias and cardiac failure. Unfortunately, there are limited therapeutic options and dialysis and kidney transplantation are still the only viable options to substitute for declining kidney function at late stage of the disease. Over the years the molecular mechanisms underlying cyst formation and progression have become better understood. Yet, this has not been paralleled by the development of novel therapeutic options. While the underlying reasons are multifold, a strategic shift is needed to accelerate therapeutics development for ADPKD. As such we hypothesize that a human-based ex vivo system using adult kidney tubule/collecting duct cells, exposed to fluid flow and with high throughput abilities will best tackle this large unmet need. To develop such a novel high-throughput screening approach we propose the following two Aims: In Aim 1 we will develop utilizing genetically engineered primary human urinary-derived kidney tubuloids, a 96-well based flow chamber and an automated analysis pipeline. The development of a human-based model that consists of adult fully, differentiated kidney epithelial cells of the different nephron segments and the collecting duct under flow conditions and operational in a high-throughput screening manner will have wide ranging applications. While the short-term aims are to utilize it towards developing ADPKD therapeutics, the setup and the data analysis pipeline can be used for any approach studying adult human kidney cells. Thus, in the long-term it is applicable towards a wide range of screening approaches including e.g., Crispr/CAS9-based genetic editing screens.

常染色体显性多囊性肾脏疾病（ADPKD）是一种遗传遗传的多器官疾病，其特征是肾脏中液体填充囊肿和细胞间纤维化的形成。这导致肾功能逐渐下降，最终导致肾衰竭。重要的是，心血管并发症是ADPKD患者发病和死亡的主要原因。这是肾素 - 血管紧张素 - 醛固酮系统过多激活的直接结果，该系统会导致高血压并增加患冠心病，心律不齐和心脏衰竭的风险。不幸的是，治疗选择有限，透析和肾脏移植仍然是唯一可以替代疾病后期肾功能下降的可行选择。多年来，囊肿形成和进展的分子机制已经得到了更好的理解。但是，这与新型治疗选择的发展尚未平行。尽管根本的原因是多重的，但仍需要进行战略转变，以加速ADPKD的疗法开发。因此，我们假设使用成年肾小管/收集管细胞，暴露于流体流动和具有较高吞吐量的人体基于人体的离体系统，最好最好地解决这一巨大的未满足需求。为了开发这种新型的高通量筛选方法，我们提出了以下两个目的：在AIM 1中，我们将利用基因工程的原代人尿液衍生的肾小管，一个基于96孔的基于96孔的流动室和自动化分析管道。基于人类的模型的开发由不同肾单位段的成人完全分化的肾上皮细胞组成，以及在流动条件下的收集导管，并以高通量筛选方式运行，将具有广泛的范围应用。尽管短期目的是利用它来开发ADPKD治疗剂，但可以将设置和数据分析管道用于研究成年人类肾细胞的任何方法。因此，从长远来看，它适用于广泛的筛选方法，包括基于CRISPR/CAS9的基因编辑筛选。