Identification of Tracheobronchial Basal Cell-Specific Genes

气管支气管基底细胞特异性基因的鉴定

• 批准号: 8686942
• 负责人: Susan D Reynolds
• 金额: $ 23.01万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8686942
• 关键词: A Mouse; Alleles; Basal Cell; Cell Differentiation process; Cell Separation; Cell physiology; Cells; Cessation of life; Characteristics; Cytometry; Development; Epithelium; Equilibrium; Esophageal; Exhibits; Figs - dietary; Future; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Genetic Models; Goals; Healed; Health; Human; In Situ Hybridization; Keratin; Life; Lung; Lung diseases; Metabolism; Methods; Modeling; Modification; Molecular; Mus; PECAM1 gene; PTPRC gene; Phenotype; Polymerase Chain Reaction; Reverse Transcription; Role; Signal Pathway; Signal Transduction; Stem cells; Testing; Thromboplastin; Tissues; Tracheobronchial; Transgenes; cell type; design; healing; human tissue; in vivo; premature; prevent; progenitor; promoter; public health relevance; respiratory; tool

DESCRIPTION (provided by applicant): The tracheobronchial epithelium (TBE) is the respiratory compartment that is maintained by the basal cell progenitor. Basal cell progenitor proliferation and differentiation are critical components of lung health and changes in these functions contribute to respiratory disease. We suggest that treatments which normalize basal cell progenitor functions can be used to heal the TBE. To achieve this goal, we must identify the signals that regulate TBE basal cell functions. This proposal is designed to resolve the issues that prevent modeling of human TBE basal cells in mice and use of genetically-modified mice to test signaling pathway function in vivo. We and others have shown that there are subsets of human and mouse TBE basal cells and that TBE basal cells are different from basal cells that maintain non-respiratory tissues. Thus, we hypothesize that: Tracheobronchial basal cells are defined by a unique gene expression signature. Aim 1: Determine the gene expression profile of human and mouse TBE basal cells. Sub-hypothesis: TBE basal cell subtypes exhibit a characteristic gene expression signature. We will use gene expression profiling (GEP) to determine if: 1) the normal human and mouse TBE is populated by molecularly-distinct subsets of basal cells; and 2) there are analogous human and mouse TBE basal cell subsets. Aim 2: Identify TBE basal cell-specific genes. Sub-hypothesis: TBE basal cells are a molecularly-distinct tissue-specific basal cell subtype. We will use GEP to determine if the mouse TBE basal cell transcriptome contains a signature that is distinct from basal cells in other tissues. We will then use quantitative reverse transcription polymerase chain reaction (qRT-PCR) and in situ hybridization analysis to identify a gene(s) that is unique to the TBE basal cell and its putative subtypes. In the future, we will create a new genetic model(s) that uses a TBE basal cell-specific promoter to modify gene expression in the mouse TBE basal cell subtype(s) that is analogous to the human TBE basal cell subtype(s). This cell type-specific approach will prevent the non-respiratory, life-threatening genetic modifications that currently confound in vivo analysis of signaling in TBE basal cells.

描述（由申请人提供）：气管支气管上皮（TBE）是由基底细胞祖细胞维持的呼吸室。基底细胞祖细胞增殖和分化是肺部健康的关键成分，这些功能的变化会导致呼吸道疾病。我们建议可以使用标准化基础细胞祖细胞功能的治疗方法来治愈TBE。为了实现此目标，我们必须确定调节基础细胞功能的信号。该建议旨在解决防止小鼠中人类TBE基础细胞建模的问题，并使用遗传改性的小鼠在体内测试信号通路的功能。我们和其他人表明，有人类和小鼠的基底细胞子集，基底细胞与维持非呼吸组织的基底细胞不同。因此，我们假设：气管支气管基底细胞由独特的基因表达特征定义。目标1：确定人和小鼠基底细胞的基因表达谱。亚刺：基底细胞亚型表现出特征性的基因表达特征。我们将使用基因表达分析（GEP）来确定是否：1）正常的人和小鼠TBE由基底细胞的分子赋予子集填充； 2）有类似的人和小鼠基底细胞子集。 AIM 2：确定基础细胞特异性基因。亚杂种：基底细胞是分子缺陷的组织特异性基底细胞亚型。我们将使用GEP来确定小鼠基底细胞转录组是否包含与其他组织中基底细胞不同的特征。我们将 然后，使用定量逆转录聚合酶链反应（QRT-PCR）和原位杂交分析来识别TBE基底细胞及其推定亚型独有的基因。将来，我们将创建一种新的遗传模型，该模型使用TBE基础细胞特异性启动子修改小鼠TBE基底细胞亚型中的基因表达，该基础细胞亚型类似于人类基底细胞亚型（S）。这种细胞类型特异性方法将防止非呼吸，威胁生命的遗传修饰，这些修饰当前在基础细胞中信号传导的体内分析中混淆。