Generation of lung progenitors from Cystic Fibrosis iPS cells

囊性纤维化 iPS 细胞产生肺祖细胞

• 批准号: 8302501
• 负责人: JAYARAJ RAJAGOPAL
• 金额: $ 24.31万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8302501
• 关键词: Biological; Biological Assay; Cell Differentiation process; Cell Line; Cell Transplantation; Cells; Chemicals; Communities; Cystic Fibrosis; Data; Development; Disease; Disease model; Embryo; Embryonic Development; Epithelial; Epithelial Cells; Epithelium; Event; Failure; Fibroblasts; Future; Generations; Genetic; Growth Factor; Hereditary Disease; Human; Hyperplasia; In Vitro; Individual; Injury; Intraepithelial Neoplasia; Laboratories; Left; Libraries; Lung; Lung diseases; Medical Research; Messenger RNA; Metaplasia; Methods; Modeling; Molecular; Morbidity - disease rate; Mucous body substance; Mus; Natural regeneration; Organogenesis; Patients; Phenotype; Pluripotent Stem Cells; Preclinical Drug Evaluation; Protocols documentation; Pulmonary Cystic Fibrosis; Reporting; Research; Respiration Disorders; Screening procedure; Signal Pathway; Signal Transduction; Site; Skin; Stem cells; Structure of parenchyma of lung; Subfamily lentivirinae; System; Testing; Transplantation; Universities; airway epithelium; base; cell type; cystic fibrosis patients; embryonic stem cell; human disease; human embryonic stem cell; in vitro Model; in vivo; induced pluripotent stem cell; interest; lung development; meetings; mortality; mouse model; new technology; non-genetic; novel; novel strategies; patient population; prevent; progenitor; regenerative therapy; research study; stem; success; transcription factor

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. PUBLIC HEALTH RELEVANCE: The discovery of human induced-pluripotent stem cells has resulted in an unprecedented opportunity to produce patient-specific cells types that can be used to generate in vitro models of human lung disease and eventually be used for human transplantation. Although induced- pluripotent stem cell lines from patients with lung disease are currently being produced, the major obstacle preventing the actual development of regenerative therapies using these cells is our inability to efficiently convert pluripotent stem cells into lun cells. This proposal aims to develop efficient and reproducible protocols to produce lung cells from stem cells.

描述（由申请人提供）：两个气道的疾病是全球死亡和发病的主要原因之一。它们通常以肺上皮的异常为特征，包括上皮增生、粘液化生、上皮发育不良和损伤后上皮再生的失败。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，缺乏人类肺部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来产生肺上皮细胞在体外的数量适合于疾病建模，药物筛选和移植。ES细胞由于其在培养中无限增殖的独特能力和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）的发现现在开辟了产生大量分化的人肺细胞的可能性，这些细胞来源于iPS细胞，而iPS细胞又是由患有各种特定肺部疾病的患者的皮肤成纤维细胞产生的。这项新技术为体外模拟人类疾病机制和开发患者特异性疗法创造了机会。然而，试图在体外有效地产生大量同质的、分化的肺上皮细胞的尝试已经遇到了有限的成功。在本申请中，我们概述了优化从人囊性纤维化iPS细胞体外生成肺祖细胞的提议。与以前的报道相反，我们将集中在表达转录因子Nkx2.1的早期肺上皮祖细胞的产生上。Nkx2.1是肺上皮祖细胞的最早和最特异的标志物，并且也在大多数成熟肺上皮细胞中表达。为了有效地产生Nkx2.1阳性的早期肺 祖细胞我们建议使用合理的基于发育的分化策略的组合，该策略概括了正常胚胎发生期间肺祖细胞的逐步分化和使用化学和生物文库的高通量无偏分化筛选。在未来，类似的策略可以用于从任何呼吸系统疾病患者中产生任何特定的肺上皮细胞群，但作为原则的证明，我们用肺最常见的遗传性疾病之一囊性纤维化来证明。 公共卫生相关性：人类诱导多能干细胞的发现带来了前所未有的机会，可以产生患者特异性细胞类型，这些细胞类型可用于产生人类肺部疾病的体外模型，并最终用于人类移植。虽然目前正在生产来自肺病患者的诱导多能干细胞系，但阻止使用这些细胞的再生疗法的实际发展的主要障碍是我们无法有效地将多能干细胞转化为lun细胞。该提案旨在开发有效和可重复的方案，从干细胞中产生肺细胞。