iPSC Modeling of the Role of NKX2-1 in Human Lung Development and Disease

描述（由申请人提供）：最近诱导多能干细胞（iPSCs）的发现为体外模拟人类疾病和更好地了解人类发育提供了前所未有的机会。iPSC体外模型系统的临床翻译是本项目的目标，其目的是了解人类发育的最早阶段，重点研究已知最早的肺命运标志物，关键转录调节因子NKX2.1在发育和人类疾病中的作用。对于这些研究，我们将利用我们开发的重编程技术，使我们能够建立一个综合的iPSCs库，这些iPSCs是从肺部谱系规范的最早时刻活跃的基因突变（如NKX2.1）引起的肺部疾病的儿童中产生的。这些疾病特异性细胞系提供了自然发生的人类模型，用于研究人类中半形、突变或显性负基因表达的影响，为了解控制正常与异常早期肺发育的机制提供了令人兴奋的机会。最终，对肺部发育生物学的详细了解将有助于更好地治疗肺部疾病患者，本项目所研究的儿童就是一个例子，即4名因NKX2.1编码区突变而具有不同临床表型的个体。我们将把我们的体外研究和模型与从这些儿童的肺组织中建立的微阵列联系起来，我们将检验我们的假设：体外iPSC系统可以应用于建模和理解遗传性肺部疾病，例如由NKX2.1活性降低或增强引起的肺-甲状腺-脑临床表型不同的患者。目的1提出定义正常与疾病特异性的人类肺上皮祖细胞分化的遗传特征，并评估这些体外模型系统如何与这些细胞来源的患者观察到的临床表型进行比较。Aim 2评估分子机制，如NKX2.1在这些患者中与疾病发病机制相关的结合或失活改变，Aim 3测试机械驱动的潜在治疗方法，以改善NKX2.1相关的人类疾病表型。

DESCRIPTION (provided by applicant): The recent discovery of induced pluripotent stem cells (iPSCs) has provided unprecedented opportunities to model human disease in vitro and better understand human development. The clinical translation of the iPSC in vitro model system is the goal of this project, whose aim is to learn about the earliest stages of human development, focused on the role in development and human disease of the earliest known marker of lung fate, the key transcriptional regulator NKX2.1. For these studies we will utilize reprogramming technology we have developed that has allowed us to establish a comprehensive bank of iPSCs generated from children with lung diseases caused by mutations in genes (such as NKX2.1) that are active from the earliest moments of lung lineage specification. These disease-specific cell lines provide naturally occurring human models of the effects of hypomorphic, mutant, or dominant negative gene expression in human beings, providing exciting opportunities to understand mechanisms that control normal vs. aberrant early lung development. Ultimately a detailed understanding of lung developmental biology should result in better treatments for patients with lung disease, exemplified by the children to be studied in this project, namely 4 individuals with divergent clinical phenotypes resulting from mutations in the coding region of NKX2.1. We will correlate our in vitro studies and models with microarrays established from the lung tissue of these children and we will test our hypothesis: the in vitro iPSC system can be applied to model and understand genetic lung diseases, exemplified by patients with divergent lung-thyroid-brain clinical phenotypes arising from reduced or augmented activity of NKX2.1. Aim 1 proposes to define the normal vs. disease-specific genetic signatures of differentiating human lung epithelial progenitors and to assess how these in vitro model systems compare to the clinical phenotypes observed in the patients from whom these cells are derived. Aim 2 evaluates the molecular mechanism, such as binding or inactivating alterations of NKX2.1 responsible for disease pathogenesis in these patients, and Aim 3 tests mechanistically-driven potential therapies for ameliorating the NKX2.1-related human disease phenotypes.