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- Title
Gene-Edited Human Kidney Organoids Reveal Mechanisms of Disease in Podocyte Development.
- Authors
Kim, Yong Kyun; Refaeli, Ido; Brooks, Craig R.; Jing, Peifeng; Gulieva, Ramila E.; Hughes, Michael R.; Cruz, Nelly M.; Liu, Yannan; Churchill, Angela J.; Wang, Yuliang; Fu, Hongxia; Pippin, Jeffrey W.; Lin, Lih Y.; Shankland, Stuart J.; Vogl, A. Wayne; McNagny, Kelly M.; Freedman, Benjamin S.
- Abstract
A critical event during kidney organogenesis is the differentiation of podocytes, specialized epithelial cells that filter blood plasma to form urine. Podocytes derived from human pluripotent stem cells (hPSC-podocytes) have recently been generated in nephron-like kidney organoids, but the developmental stage of these cells and their capacity to reveal disease mechanisms remains unclear. Here, we show that hPSC-podocytes phenocopy mammalian podocytes at the capillary loop stage (CLS), recapitulating key features of ultrastructure, gene expression, and mutant phenotype. hPSC-podocytes in vitro progressively establish junction-rich basal membranes (nephrin+podocin+ZO-1+) and microvillus-rich apical membranes (podocalyxin+), similar to CLS podocytes in vivo. Ultrastructural, biophysical, and transcriptomic analysis of podocalyxin-knockout hPSCs and derived podocytes, generated using CRISPR/Cas9, reveals defects in the assembly of microvilli and lateral spaces between developing podocytes, resulting in failed junctional migration. These defects are phenocopied in CLS glomeruli of podocalyxin-deficient mice, which cannot produce urine, thereby demonstrating that podocalyxin has a conserved and essential role in mammalian podocyte maturation. Defining the maturity of hPSC-podocytes and their capacity to reveal and recapitulate pathophysiological mechanisms establishes a powerful framework for studying human kidney disease and regeneration. S tem C ells 2017;35:2366-2378
- Publication
Stem Cells, 2017, Vol 35, Issue 12, p2366
- ISSN
1066-5099
- Publication type
Article
- DOI
10.1002/stem.2707