Generation of lung progenitors from Cystic Fibrosis iPS cells

描述（由申请人提供）：双呼吸道疾病是世界范围内死亡率和发病率的主要原因之一。它们通常以肺上皮异常为特征，包括上皮增生、粘膜化生、上皮发育不良和损伤后上皮再生失败。小鼠模型不能反映人类肺部疾病的许多最重要的特征。此外，缺乏人体肺组织是阻碍分析导致人类肺部疾病的细胞和分子机制的关键瓶颈之一。这些问题需要新的方法在体外产生适合疾病建模、药物筛选和移植的肺上皮细胞。胚胎干细胞因其在培养中无限增殖的独特能力以及产生所有分化细胞类型的能力而引起了研究和医学界的极大兴趣。诱导多能干细胞（iPS细胞）的发现使得从iPS细胞中产生大量分化的人类肺细胞成为可能，而iPS细胞又是从患有各种特定肺部疾病的患者的皮肤成纤维细胞中产生的。这项新技术创造了在体外模拟人类疾病机制和开发针对患者的治疗方法的机会。然而，在体外有效地产生大量同质、分化的肺上皮细胞的尝试取得了有限的成功。在这个应用中，我们概述了一个建议，以优化体外生成肺祖细胞从人囊性纤维化iPS细胞。与之前的报道相反，我们将重点关注表达转录因子Nkx2.1的早期肺上皮祖细胞的产生。Nkx2.1是最早、最特异的肺上皮祖细胞标志物，在大多数成熟肺上皮细胞中也有表达。高效生成nkx2.1阳性早期肺

DESCRIPTION (provided by applicant): Diseases of both the airways are amongst the leading causes of mortality and morbidity worldwide. They are often characterized by abnormalities of the lung epithelium including epithelial hyperplasia, mucous metaplasia, epithelial dysplasia and the failure of epithelial regeneration after injury. Mouse models do not reflect many of the most important features of human lung disease. In addition, the lack of human lung tissue is one of the key bottlenecks preventing an analysis of the cellular and molecular mechanisms that result in human lung disease. These problems require new approaches to generate lung epithelial cells in vitro in numbers suitable for disease modeling, drug screening and transplantation. ES cells have attracted significant interest in research and medical communities due to their unique capacity for unlimited proliferation in culture and their ability to give rise to all differentiate cell types. The discovery of induced-pluripotent stem cells (iPS cells) now opens up the possibility of generating large numbers of differentiated human lung cells derived from iPS cells which are turn are created from skin fibroblasts from patients with a variety of specific lung disorders. Thi new technology creates an opportunity to model human disease mechanisms in vitro and to develop patient-specific therapies. However, attempts to efficiently produce large numbers of homogenous, differentiated lung epithelial cells in vitro have met with limited success. In this application we outline a proposal to optimize the in vitro generation of lung progenitors from human Cystic Fibrosis iPS cells. In contrast to previous reports, we will focus on the generation of early lung epithelial progenitors expressing the transcription factor Nkx2.1. Nkx2.1 is the earliest and the most specific marker of pulmonary epithelial progenitors and is also expressed in the majority of mature lung epithelial cells. To efficiently generate Nkx2.1-positive early lung progenitors we propose to use a combination of rational development-based differentiation strategies that recapitulates the stepwise differentiation of lung progenitor cells during normal embryogenesis and high throughput unbiased differentiation screens using chemical and biological libraries. In the future, a similar strategy can be used to produce any specific lung epithelial population from patients with any respiratory disorder, but as proof of principle we stat with one of the most common genetic disorders of the lung, Cystic Fibrosis.