Gene-Edited Human Kidney Organoids Reveal Mechanisms of Disease in Podocyte Development. (29th September 2017)

• Record Type: Journal Article
• 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.
• Is Part Of: Stem cells. Volume 35:Number 12(2017:Dec.)
• Page Start: 2366
• Page End: 2378
• Publication Date: 2017-09-29
• Subjects: 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; Cloning -- Periodicals; Clone cells -- Periodicals; Stem cells -- Periodicals; Cell Differentiation -- Periodicals; Cell Division -- Periodicals; Clone Cells -- Periodicals; Hematopoietic Stem Cells -- Periodicals; Stem Cells -- Periodicals; 571.84
• DOI: 10.1002/stem.2707
• Languages: English
• ISSNs: 1066-5099

Abstract

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.