Generation of Human PSC-Derived Kidney Organoids with Patterned Nephron Segments and a De Novo Vascular Network

Human pluripotent stem cells-derived kidney organoids recapitulate developmental processes and tissue architectures, but intrinsic limitations, such as a lack of vasculature and functionality, have greatly hampered their application. Here we establish a versatile protocol for generating vascularized three-dimensional kidney organoids. We employ dynamic modulation of WNT signaling to control the relative proportion of proximal versus distal nephron segments, thereby producing a correlative level of VEGFA to define a resident vascular network. Single cell RNA-sequencing identifies a subset of nephron progenitor cells as a potential source of renal vasculature. These kidney organoids undergo further structural and functional maturation upon implantation. Using this kidney organoid platform, we establish an in vitro model of autosomal recessive polycystic kidney disease, the cystic phenotype of which can be effectively prevented by gene correction or drug treatment. Our studies provide new avenues for studying human kidney development, modeling disease pathogenesis, and performing patient-specific drug validation.

人源多能干细胞衍生的肾脏类器官能够重现发育过程和组织结构，然而，固有的局限性，例如血管和功能的缺失，极大地阻碍了它们的应用。在本研究中，我们建立了一种通用的方法，用于生成血管化的三维肾脏类器官。我们通过动态调节WNT信号通路，控制近端与远端肾小管段的比例，从而产生相应的VEGFA水平，以定义驻留的血管网络。单细胞RNA测序鉴定出肾小管祖细胞亚群作为肾脏血管的潜在来源。这些肾脏类器官在植入后进一步经历结构和功能的成熟。利用这一肾脏类器官平台，我们建立了一种自发性常染色体隐性多囊肾病体外模型，其囊肿表型可以通过基因校正或药物治疗有效预防。我们的研究为研究人类肾脏发育、疾病发病机制建模以及进行患者特异性药物验证提供了新的途径。