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

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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.