McCauley Katherine B, Hawkins Finn, Serra Maria, Thomas Dylan C, Jacob Anjali, Kotton Darrell N
Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
Center for Regenerative Medicine of Boston University and Boston Medical Center, Boston, MA 02118, USA.
Cell Stem Cell. 2017 Jun 1;20(6):844-857.e6. doi: 10.1016/j.stem.2017.03.001. Epub 2017 Mar 30.
Effective derivation of functional airway organoids from induced pluripotent stem cells (iPSCs) would provide valuable models of lung disease and facilitate precision therapies for airway disorders such as cystic fibrosis. However, limited understanding of human airway patterning has made this goal challenging. Here, we show that cyclical modulation of the canonical Wnt signaling pathway enables rapid directed differentiation of human iPSCs via an NKX2-1 progenitor intermediate into functional proximal airway organoids. We find that human NKX2-1 progenitors have high levels of Wnt activation but respond intrinsically to decreases in Wnt signaling by rapidly patterning into proximal airway lineages at the expense of distal fates. Using this directed approach, we were able to generate cystic fibrosis patient-specific iPSC-derived airway organoids with a defect in forskolin-induced swelling that is rescued by gene editing to correct the disease mutation. Our approach has many potential applications in modeling and drug screening for airway diseases.
从诱导多能干细胞（iPSC）有效衍生出功能性气道类器官，将为肺部疾病提供有价值的模型，并促进针对诸如囊性纤维化等气道疾病的精准治疗。然而，对人类气道模式形成的了解有限，使得这一目标具有挑战性。在这里，我们表明，经典Wnt信号通路的周期性调节能够通过NKX2-1祖细胞中间体，使人类iPSC快速定向分化为功能性近端气道类器官。我们发现，人类NKX2-1祖细胞具有高水平的Wnt激活，但通过以远端命运为代价快速分化为近端气道谱系，对Wnt信号的降低具有内在反应。使用这种定向方法，我们能够生成囊性纤维化患者特异性的iPSC衍生气道类器官，其在福司可林诱导的肿胀方面存在缺陷，通过基因编辑纠正疾病突变可挽救该缺陷。我们的方法在气道疾病的建模和药物筛选中有许多潜在应用。
