Lee Kong Chian School of Medicine, Nanyang Technological University Singapore, 11 Mandalay Road, Singapore 308232, Singapore.
Lee Kong Chian School of Medicine, Nanyang Technological University Singapore, 11 Mandalay Road, Singapore 308232, Singapore; Human Genetics, Genome Institute of Singapore, A(∗)STAR, Singapore 138672, Singapore.
Cell Stem Cell. 2019 Sep 5;25(3):373-387.e9. doi: 10.1016/j.stem.2019.06.009. Epub 2019 Jul 11.
Human pluripotent stem cell-derived kidney organoids recapitulate developmental processes and tissue architecture, but intrinsic limitations, such as lack of vasculature and functionality, have greatly hampered their application. Here we establish a versatile protocol for generating vascularized three-dimensional (3D) kidney organoids. We employ dynamic modulation of WNT signaling to control the relative proportion of proximal versus distal nephron segments, producing a correlative level of vascular endothelial growth factor A (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 (ARPKD), 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.
人多能干细胞衍生的肾类器官可重现发育过程和组织架构,但由于内在的局限性,如缺乏脉管系统和功能,极大地限制了它们的应用。在这里,我们建立了一个通用的方案来生成血管化的三维(3D)肾类器官。我们采用 WNT 信号的动态调节来控制近端和远端肾单位段的相对比例,产生相应水平的血管内皮生长因子 A(VEGFA)来定义固有血管网络。单细胞 RNA 测序确定了一组肾祖细胞作为肾脉管系统的潜在来源。这些肾类器官在植入后进一步进行结构和功能成熟。利用这个肾类器官平台,我们建立了常染色体隐性多囊肾病(ARPKD)的体外模型,其囊性表型可以通过基因矫正或药物治疗有效地预防。我们的研究为研究人类肾脏发育、疾病发病机制建模和进行患者特异性药物验证提供了新途径。