Open AccessGene‐Edited Human Kidney Organoids Reveal Mechanisms of Disease in Podocyte Development
Author(s) -
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.
Publication year - 2017
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.2707
Subject(s) - podocalyxin , podocyte , biology , microbiology and biotechnology , nephrin , induced pluripotent stem cell , nephron , kidney , endocrinology , embryonic stem cell , genetics , gene , proteinuria
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