Journal
STEM CELLS
Volume 35, Issue 12, Pages 2366-2378Publisher
OXFORD UNIV PRESS
DOI: 10.1002/stem.2707
Keywords
Pluripotent stem cells; Kidney; Adhesion receptors; Differentiation; Gene targeting; Developmental biology; Focal segmental glomerulosclerosis; Nephrogenesis; Cell adhesion; Foot processes; Slit diaphragm; Podocalyxin; Nephrin; Podocin; Genome editing; Biophysics
Categories
Funding
- NIH National Institute of Diabetes and Digestive and Kidney Diseases [K01DK102826, R01DK097598, UH2DK107343]
- National Heart, Lung, and Blood Institute [K25HL135432]
- National Institute of General Medical Sciences [P01GM081619]
- Eunice Kennedy Shriver National Institute of Child Health and Human Development [R24HD000836]
- National Kidney Foundation Young Investigator Grant
- American Society of Nephrology Carl W. Gottschalk Research Scholar Award
- Natural Sciences and Engineering Research Council of Canada [RGPIN-155397-13]
- Directorate for Biological Sciences, National Science Foundation [IDBR DBI-1353718]
- Direct For Biological Sciences
- Div Of Biological Infrastructure [1353718] Funding Source: National Science Foundation
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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. Stem Cells2017;35:2366-2378
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