Development of high-throughput platforms for human kidney disease modeling.

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Novel human in vitro model systems for kidney diseases are urgently needed since data from rodent models is often not transferable to humans. Furthermore, in vitro platforms can be scaled-up toward true high-throughput and thus will advance target discovery and validation as well as drug screening studies. In this project, we will develop novel in vitro model systems by combining our cell-biology and bio-engineering expertise. 1) We will develop a macroscopic model of a perfused human kidney tubule using novel adult human kidney organoid cells and model adult polycystic kidney disease using CRISPR /Cas9 gene editing in this system. 2) We will then move towards a real resolution kidney system using rod shaped microgels with a scalable diameter of 8-200 micrometers for anisogel alignment. For this, we will use novel adult kidney immortalized cell lines (pericytes, endothelium, epithelium) and induce tubule injury by chemical exposure or genetic engineering with the aim to develop a model of kidney fibrosis. 3) Our final aim is to move the system towards true high-throughput disease modeling and we will achieve this using a novel bioprinting technique. We will employ a new synthetic polyethylene glycole (PEG)-based bioink, consisting of rod-shaped microgels, to model the human tubulo-interstitium in 96 and 348 well plates. Fibrosis and ADPKD will be induced as described above. We will compare these in vitro platforms with human kidney disease specimen and donor biopsies using scRNA-sequencing with the goal to have a model that reflects human disease.