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

由于啮齿类动物模型的数据通常不能转移到人类，因此迫切需要新型的肾脏疾病人类体外模型系统。此外，体外平台可以扩大到真正的高通量，从而将推进靶点发现和验证以及药物筛选研究。在这个项目中，我们将结合我们的细胞生物学和生物工程专业知识开发新的体外模型系统。1)我们将使用新的成人肾类器官细胞开发灌注的人肾小管的宏观模型，并在该系统中使用CRISPR /Cas9基因编辑建立成人多囊肾病模型。2)然后，我们将使用直径为8-200微米的棒状微凝胶进行异向凝胶对齐，从而实现真实的分辨率肾脏系统。为此，我们将使用新的成人肾脏永生化细胞系（周细胞、内皮细胞、上皮细胞），并通过化学暴露或基因工程诱导肾小管损伤，目的是开发肾纤维化模型。3)我们的最终目标是将系统推向真正的高通量疾病建模，我们将使用一种新的生物打印技术来实现这一目标。我们将采用一种新的合成聚乙二醇（PEG）为基础的生物墨水，由棒状微凝胶，在96和348孔板中模拟人肾小管上皮细胞。将如上所述诱导纤维化和ADPKD。我们将使用scRNA测序将这些体外平台与人类肾脏疾病标本和供体活检进行比较，目的是建立一个反映人类疾病的模型。

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.