Interrogation of individual cells to identify progenitor cells and their niches

询问单个细胞以识别祖细胞及其生态位

• 批准号: 8114057
• 负责人: MARK A KRASNOW
• 金额: $ 101.61万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8114057
• 关键词: Animal Model; Cells; Cellular biology; Color; Complex; Development; Disease; Drosophila genus; Gene Expression; Genetic; Genomics; Human; In Situ Hybridization; In Vitro; Individual; Libraries; Lung; Lung diseases; Maintenance; Mesenchyme; Methods; Microfluidics; Microscopic; Microscopy; Molecular Profiling; Mouse Strains; Mus; Pathway interactions; Pattern; Population; Procedures; Resolution; Signal Pathway; Signal Transduction; Signaling Molecule; Stem cells; Structure of parenchyma of lung; Techniques; Tissues; Transgenic Mice; in vivo; lung development; molecular marker; prevent; progenitor; research study; response; three dimensional structure; tool

DESCRIPTION (provided by applicant): Identifying and characterizing lung progenitor cells and their signaling niches is crucial for understanding how a healthy lung is built and maintained, how alteration of development and maintenance pathways cause or contribute to lung disease, and how disease can be prevented and damaged lung tissue restored or replaced. Identifying and characterizing lung progenitor cells and their niches has been hampered by the complex three-dimensional structure of the lung and the lack of tools to mark, follow the fate, and manipulate gene expression in individual lung cells in vivo. Here we propose to develop such tools, by adapting for use in mouse lung the systematic genetic approaches and single cell resolution genetic tools ("clonal analysis") that have been used over the past decade to elucidate progenitor and stem cells and their signaling niches in the model organism Drosophila. To facilitate this, we will develop a new microscopy procedure (multidimensional microscopic molecular profiling) that allows the levels of dozens of molecular markers to be quantified at subcellular resolution on a single tissue section, the equivalent of multi-dimensional FACS analysis for intact tissues. We combine these in vivo approaches with a high throughput in vitro approach that takes advantage of recent advances in genomics and microfluidics to characterize individual lung progenitor cells and their developmental responses to the signals identified in the in vivo experiments. This combined approach is general and applicable to progenitor cells throughout the lung and other mouse tissues, although we focus on the poorly characterized progenitor cells in lung mesenchyme.

描述（由申请人提供）： 识别和表征肺祖细胞及其信号壁ches对于了解健康肺的建立和维护，发育和维持途径的改变如何引起或导致肺部疾病以及如何预防和疾病恢复或替换疾病至关重要。肺部及其壁细胞的识别和表征受到肺的复杂三维结构的阻碍，缺乏标记，遵循命运的工具，并在体内单个肺细胞中操纵基因表达。在这里，我们建议开发这种工具，通过适应小鼠肺部使用系统的遗传方法和单细胞分辨率遗传工具（“克隆分析”），这些工具已在过去十年中用于阐明祖细胞和干细胞及其在模型有机体果蝇中的信号壁细胞。为了促进这一点，我们将开发一种新的显微镜程序（多维显微镜分子分析），该程序允许在单个组织部分的亚细胞分辨率上量化数十个分子标记的水平，这是完整组织的多维面部分析的等效性。我们将这些体内方法与高通量的体外方法结合在一起，该方法利用了最近的基因组学和微富集术的进步来表征单个肺祖细胞及其对体内实验中鉴定的信号的发育反应。这种组合的方法是一般的，适用于整个肺部和其他小鼠组织的祖细胞，尽管我们专注于肺间充质中表征较差的祖细胞。