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: 8442861
• 负责人: JAYARAJ RAJAGOPAL
• 金额: $ 18.94万
• 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/8442861
• 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; 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; screening; 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.

描述(申请人提供)：这两种呼吸道的疾病都是世界范围内死亡和发病的主要原因之一。通常以肺上皮异常为特征，包括上皮性增生、粘液化生、上皮异型增生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，缺乏人类肺组织是阻碍对导致人类肺部疾病的细胞和分子机制进行分析的关键瓶颈之一。这些问题需要新的方法在体外产生适合于疾病建模、药物筛选和移植的肺上皮细胞。由于ES细胞在培养过程中具有无限增殖的独特能力，并且能够分化为各种类型的细胞，因此在研究和医学界引起了极大的兴趣。诱导多能干细胞(iPS细胞)的发现为从iPS细胞产生大量分化的人类肺细胞提供了可能，而iPS细胞又是从各种特定肺部疾病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，在体外高效地产生大量同质的、分化的肺上皮细胞的尝试遇到了有限的成功。在这项应用中，我们概述了一项优化人囊性纤维化iPS细胞体外培养肺祖细胞的建议。与以前的报道不同，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生成。Nkx2.1是肺上皮祖细胞最早和最特异的标志物，在大多数成熟的肺上皮细胞中也有表达。高效地产生Nkx2.1阳性的早期肺 我们建议使用基于合理发育的分化策略和高通量、无偏倚的分化筛选相结合的方法，利用化学和生物文库进行高通量、无偏见的分化筛选。在未来，类似的策略可以用来从任何呼吸系统疾病的患者身上产生任何特定的肺上皮细胞群，但作为原理的证据，我们统计了肺部最常见的遗传性疾病之一，囊性纤维化。