Department of Surgery, University of Wisconsin Madison, Wisconsin Institute for Medical Research, Madison, WI, USA.
Department of Biostatistics & Medical Informatics, University of Wisconsin Madison, Madison, WI, USA.
Nat Commun. 2019 Sep 24;10(1):4161. doi: 10.1038/s41467-019-12069-w.
Development of treatments for vocal dysphonia has been inhibited by lack of human vocal fold (VF) mucosa models because of difficulty in procuring VF epithelial cells, epithelial cells' limited proliferative capacity and absence of cell lines. Here we report development of engineered VF mucosae from hiPSC, transfected via TALEN constructs for green fluorescent protein, that mimic development of VF epithelial cells in utero. Modulation of FGF signaling achieves stratified squamous epithelium from definitive and anterior foregut derived cultures. Robust culturing of these cells on collagen-fibroblast constructs produces three-dimensional models comparable to in vivo VF mucosa. Furthermore, we demonstrate mucosal inflammation upon exposure of these constructs to 5% cigarette smoke extract. Upregulation of pro-inflammatory genes in epithelium and fibroblasts leads to aberrant VF mucosa remodeling. Collectively, our results demonstrate that hiPSC-derived VF mucosa is a versatile tool for future investigation of genetic and molecular mechanisms underlying epithelium-fibroblasts interactions in health and disease.
由于获取人声带（VF）黏膜的上皮细胞困难、上皮细胞增殖能力有限且缺乏细胞系，因此治疗声嘶的方法一直难以发展。在此，我们报告了利用 TALEN 构建体转染的 hiPSC 构建工程化 VF 黏膜的方法，该方法模拟了 VF 上皮细胞在体内的发育过程。通过调节 FGF 信号通路，可从明确的和前肠来源的培养物中获得复层鳞状上皮。将这些细胞在胶原蛋白-成纤维细胞构建体上大量培养可产生类似于体内 VF 黏膜的三维模型。此外，我们还证明了这些构建体在暴露于 5%香烟烟雾提取物后会引发黏膜炎症。上皮细胞和成纤维细胞中促炎基因的上调导致异常的 VF 黏膜重塑。总的来说，我们的结果表明，hiPSC 衍生的 VF 黏膜是研究健康和疾病中上皮细胞-成纤维细胞相互作用的遗传和分子机制的一种多功能工具。
