Division of Cell Matrix Biology and Regenerative Medicine, Faculty of Biology, Medicine and Health, University of Manchester, and the Manchester Academic Health Science Centre, Manchester, UK.
Division of Cell Matrix Biology and Regenerative Medicine, Faculty of Biology, Medicine and Health, University of Manchester, and the Manchester Academic Health Science Centre, Manchester, UK.
Stem Cell Reports. 2021 Jan 12;16(1):134-148. doi: 10.1016/j.stemcr.2020.11.007. Epub 2020 Dec 10.
MicroRNAs (miRNAs) are gene expression regulators and they have been implicated in acquired kidney diseases and in renal development, mostly through animal studies. We hypothesized that the miR-199a/214 cluster regulates human kidney development. We detected its expression in human embryonic kidneys by in situ hybridization. To mechanistically study the cluster, we used 2D and 3D human embryonic stem cell (hESC) models of kidney development. After confirming expression in each model, we inhibited the miRNAs using lentivirally transduced miRNA sponges. This reduced the WT1 metanephric mesenchyme domain in 2D cultures. Sponges did not prevent the formation of 3D kidney-like organoids. These organoids, however, contained dysmorphic glomeruli, downregulated WT1, aberrant proximal tubules, and increased interstitial capillaries. Thus, the miR-199a/214 cluster fine-tunes differentiation of both metanephric mesenchymal-derived nephrons and kidney endothelia. While clinical implications require further study, it is noted that patients with heterozygous deletions encompassing this miRNA locus can have malformed kidneys.
微小 RNA（miRNAs）是基因表达的调控因子，它们与获得性肾脏疾病和肾脏发育有关，这主要是通过动物研究发现的。我们假设 miR-199a/214 簇调节人类肾脏发育。我们通过原位杂交检测了其在人胚肾中的表达。为了从机制上研究该簇，我们使用了二维和三维人类胚胎干细胞（hESC）肾脏发育模型。在确认每种模型中的表达后，我们使用慢病毒转导的 miRNA 海绵抑制了 miRNA。这减少了二维培养物中 WT1 后肾间充质的域。海绵并没有阻止 3D 肾类器官的形成。然而，这些类器官包含发育不良的肾小球、下调的 WT1、异常的近端小管和增加的间质毛细血管。因此，miR-199a/214 簇精细调节后肾间充质衍生的肾单位和肾脏内皮细胞的分化。虽然临床意义需要进一步研究，但值得注意的是，包含该 miRNA 基因座的杂合缺失的患者可能会出现畸形的肾脏。
