Methods for the amplification and identification of airway stem cells

描述（由申请人提供）：慢性肺部疾病具有上皮修复缺陷的共同特征，导致组织稳态丧失和肺功能进一步恶化。该提案的目标是提高我们在健康和疾病中调节组织干细胞的能力，并通过原位刺激驻留细胞或通过开发基于细胞的治疗策略来开发增强上皮再生的策略。我们最近开发并表征了小鼠，其中通过条件表达N-末端截短的稳定形式的连环蛋白，在传导气道内扩增干细胞样细胞。本申请的目的是开发一种小鼠模型，通过条件表达？N-连环蛋白该模型将用于揭示发育中和成人肺中上皮细胞分化的关键阶段，这些阶段易受稳定的β-连环蛋白的影响。目的1将建立小鼠，允许条件性调节发育中和成年肺的CCSP表达细胞内的稳定化β-连环蛋白。该模型将用于确定小鼠细支气管上皮细胞中稳定的β-连环蛋白的分化抑制特性的可逆性。目的2将确定稳定的β-连环蛋白的分化调节作用是否仅限于干细胞，或者可以更广泛地应用于调节其他祖细胞类型（Clara细胞）的分化潜力。本申请的结果将确定旨在稳定β-连环蛋白的遗传干预（以及通过外推，药理干预）是否可用作体内扩增气道干细胞的工具。这些实验是进一步开发针对内源性肺组织干细胞的基于细胞的治疗的必要先决条件。公共卫生相关性：排列在肺部气道中的上皮细胞履行在肺部疾病环境中丧失的健康中的关键功能。潜在的缺陷是上皮修复或重建正常功能的能力。本提案中进行的研究将侧重于开发损伤后修复气道上皮的新方法。目的是利用信号通路调节能够修复气道上皮的上皮细胞的数量。这些研究将为开发能够增强慢性肺病患者上皮修复和肺功能恢复的药物提供新的信息。

DESCRIPTION (provided by applicant): Chronic lung diseases share the common feature of epithelial repair defects that contribute to loss of tissue homeostasis and further deterioration of lung function. Goals of this proposal are to improve our ability regulate tissue stem cells in health and disease, and to develop strategies to enhance epithelial regeneration either through stimulation of resident cells in situ or through development of cell-based therapeutic strategies. We have recently developed and characterized mice in which stem-like cells within conducting airways are amplified through conditional expression of an N-terminally truncated stabilized form of the ¿-catenin protein. Goals of this application are to develop a mouse model for reversible in vivo stabilization of the stem cell phenotype through conditional expression of ?N¿-catenin. This model will be used to reveal critical stages of epithelial cell differentiation in the developing and adult lung that are susceptible to the effects of stabilized ¿-catenin. Aim 1 will establish mice allowing conditional regulation of stabilized ¿-catenin within CCSP- expressing cells of the developing and adult lung. This model will be used to determine the reversibility of differentiation-arresting properties of stabilized ¿-catenin in mouse bronchiolar epithelial cells. Aim 2 will determine whether the differentiation modulating effects of stabilized ¿-catenin are limited to stem cells or can be more broadly applied to modulate the differentiation potential of other progenitor cell types (Clara cells). Results from this application will determine whether genetic interventions (and by extrapolation, pharmacologic interventions) aimed at stabilizing ¿-catenin can be used as a tool for amplification of airway stem cells in vivo. These experiments are a necessary prerequisite for the further development of therapeutic interventions aimed at cell-based therapies targeted towards endogenous lung tissue stem cells. PUBLIC HEALTH RELEVANCE: Epithelial cells that line airways of the lung fulfill critical functions in health that are lost in the setting of lung disease. The underlying defect is the capacity of the epithelium to repair or to re-establish normal function. Research to be conducted in this proposal will focus on the development of new methods to repair the epithelium of airways following injury. Goals are to take advantage of signaling pathways that regulate the number of epithelial cells capable of repairing the airway epithelium. These studies will provide new information for the development of drugs capable of enhancing epithelial repair and restoration of lung function to patients with chronic lung disease.