Map the human airway basal cell niche and the role that FGFR2 plays within it

项目摘要/摘要 慢性阻塞性肺疾病(COPD)、哮喘、原发性睫状体运动障碍(PCD)和囊性纤维化 (Cf)，代表由遗传和环境因素复杂相互作用引起的呼吸道疾病。他们是一个 发病和死亡的主要原因和有效的治疗方法各不相同。哮喘和哮喘的异质性 COPD和PCD和CF的罕见给他们的研究带来了巨大的挑战。尽管他们的 发病机制这些呼吸道疾病有共同的协调反应，涉及串扰 上皮细胞、间充质细胞和免疫细胞最终导致呼吸道重塑[2]。这是最基本的 重建的是基底细胞(BCS)，它是呼吸道的主要干细胞，以及它们的壁龛[3]。遗传性小鼠模型 提示相邻间充质分泌的成纤维细胞生长因子配体与上皮性FGFR2受体结合维持BCS [4][5]。作为对损伤的反应，成纤维细胞生长因子与河马和WNT7b协调上皮-间充质信号 网络实现上皮再生[6]-[8]。FGFR2信号在人类BCS中重要吗？令人惊讶的是， 尽管这个问题很重要，但人们对此知之甚少。我们将检验以下假设：FGFR2信号 对于维持成人呼吸道基底细胞的自我更新能力是必不可少的。在目标1中，我们将 利用空间转录学确定人的气管和软骨间呼吸道的基底细胞生态位， 配体-受体信号分析和原位杂交技术。在目标2中，我们将测试FGFR2 利用人原代和诱导物维持成人呼吸道基底细胞的信号转导是必不可少的 多能干细胞来源的BCS配备了基因编辑技术。在结束这一目标时，我们 已经确定了成纤维细胞生长因子在维持BC计划中的重要性。尽管这代表着一个雄心勃勃的 项目在短时间内，工具、细胞、方案、组织样本和合作者与适当 专业知识，都已就位。这个项目的范围和目标将使我接触到各种各样的技术 适用于F32候选人。最终，这笔赠款将产生令人兴奋的、新颖的数据，我可以用它来移动 进入我职业生涯的下一个阶段，作为一名研究肺部生物学的内科科学家。

Project Summary/Abstract Chronic obstructive pulmonary disease (COPD), asthma, primary ciliary dyskinesia (PCD) and cystic fibrosis (CF), represent airway diseases caused by complex interplay of genetic and environmental factors. They are a major cause of morbidity and mortality and effective therapies are variable. The heterogeneity of asthma and COPD and the rarity of PCD and CF pose significant challenges to their study. Despite differences in their pathogenesis these airway diseases have in common coordinated responses that involve crosstalk between epithelial, mesenchymal, and immune cells that eventually leads to airway remodeling [2]. Fundamental to this remodeling are basal cells (BCs), the major stem cell of the airway, and their niche [3]. Genetic mouse models suggest that Fgf ligands secreted by adjacent mesenchyme bind to epithelial Fgfr2 receptors to maintain BCs [4][5]. In response to injury, Fgf coordinates with Hippo and Wnt7b in an epithelial-mesenchymal signaling network to achieve epithelial regeneration [6]–[8]. Is FGFR2 signaling important in human BCs? Surprisingly, despite the importance of this question, little is known. We will test the following hypothesis: FGFR2 signaling is essential in maintaining the self-renewal capability of the human adult airway basal cell. In aim 1, we will define the basal cell niche in the human trachea and intercartilagenous airways using spatial transcriptomics, ligand-receptor signaling analyses and in situ hybridization techniques. In aim 2, we will test that FGFR2 signaling is essential in maintaining human adult airway basal cells using human primary and induced pluripotent stem cells derived BCs equipped with gene editing technology. At the conclusion of this aim we have determined the importance of FGF in maintaining the BC program. Though this represents an ambitious project in short timeframe, the tools, cells, protocols, tissue samples, and collaborators with appropriate expertise, are all in place. The scope and goals of this project will expose me to a wide variety of techniques suitable for an F32 candidate. Ultimately, this grant will generate exciting, novel data with which I can move forward into the next stage of my career as a physician-scientist studying lung biology.