3D organoid-derived human glomeruli for personalised podocyte disease model ling and drug screening
3D organoid-derived human glomeruli for personalised podocyte disease model ling and drug screening
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DOI：
10.1038/s41467-018-07594-z
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发表时间：
2018-12-04
影响因子：
16.6
通讯作者：
Little, Melissa H.
中科院分区：
文献类型：
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作者：
Hale, Lorna J.;Howden, Sara E.;Little, Melissa H.
肾肾小球内的足细胞通过形成具有中间狭缝隔膜的交错足突来维持过滤屏障，其中破坏导致蛋白尿。迄今为止，对人类足细胞病的研究采用了2D培养的原代或永生化足细胞系。在这里，我们比较了从诱导多能干细胞衍生的肾类器官中筛选的3D人肾小球与条件永生化人足细胞系，揭示了与2D培养物相比，改善的足细胞特异性基因表达，体外维持极化蛋白定位和改善的肾小球基底膜基质体。类器官来源的肾小球在培养物中保留标记物表达96小时，证明适合于毒性筛选。此外，患有复合杂合NPHS1突变的先天性肾病综合征患者的3D类器官肾小球显示NEPHRIN和PODOCIN的蛋白水平降低。因此，人iPSC衍生的类器官肾小球代表了人足细胞病的体外建模和筛选足细胞毒性的一种可行方法。
The podocytes within the glomeruli of the kidney maintain the filtration barrier by forming interdigitating foot processes with intervening slit diaphragms, disruption in which results in proteinuria. Studies into human podocytopathies to date have employed primary or immortalised podocyte cell lines cultured in 2D. Here we compare 3D human glomeruli sieved from induced pluripotent stem cell-derived kidney organoids with conditionally immortalised human podocyte cell lines, revealing improved podocyte-specific gene expression, maintenance in vitro of polarised protein localisation and an improved glomerular basement membrane matrisome compared to 2D cultures. Organoid-derived glomeruli retain marker expression in culture for 96 h, proving amenable to toxicity screening. In addition, 3D organoid glomeruli from a congenital nephrotic syndrome patient with compound heterozygous NPHS1 mutations reveal reduced protein levels of both NEPHRIN and PODOCIN. Hence, human iPSC-derived organoid glomeruli represent an accessible approach to the in vitro modelling of human podocytopathies and screening for podocyte toxicity.