Gene-Edited Human Kidney Organoids Reveal Mechanisms of Disease in Podocyte Development.
Gene-Edited Human Kidney Organoids Reveal Mechanisms of Disease in Podocyte Development.
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DOI：
10.1002/stem.2707
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发表时间：
2017-12
期刊：
影响因子：
--
通讯作者：
Freedman BS
中科院分区：
文献类型：
--
作者：
Kim YK;Refaeli I;Brooks CR;Jing P;Gulieva RE;Hughes MR;Cruz NM;Liu Y;Churchill AJ;Wang Y;Fu H;Pippin JW;Lin LY;Shankland SJ;Vogl AW;McNagny KM;Freedman BS
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 gene-edited hPSCs and derived podocytes, generated using CRISPR/Cas9, reveals that podocalyxin is essential for the assembly of microvilli and lateral spaces between developing podocytes. 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. Comparison of stem cell-derived human kidney organoids in vitro with genetic mouse models in vivo reveals that podocytes (red), the filtering cells of the kidney, undergo apical-to-basal junctional migration at the capillary loop stage (CLS). A critical player in this process is podocalyxin (PODXL), which induces negatively-charged microvilli between neighboring podocytes, pushing them apart.
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影响因子：
3.7
作者：
Nielsen JS;Graves ML;Chelliah S;Vogl AW;Roskelley CD;McNagny KM
通讯作者：
McNagny KM
DOI：
10.1084/jem.194.1.13
发表时间：
2001-07-02
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Doyonnas R;Kershaw DB;Duhme C;Merkens H;Chelliah S;Graf T;McNagny KM
通讯作者：
McNagny KM
影响因子：
5.2
作者：
Choo, Andre B.;Tan, Heng Liang;Yap, Miranda
通讯作者：
Yap, Miranda
影响因子：
13.6
作者：
Ronconi, Elisa;Sagrinati, Costanza;Romagnani, Paola
通讯作者：
Romagnani, Paola
影响因子：
30.8
作者：
Boute, N;Gribouval, O;Antignac, C
通讯作者：
Antignac, C