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

绘制人类气道基底细胞生态位图以及 FGFR2 在其中发挥的作用

• 批准号: 10752101
• 负责人: Daniel J. Wallman
• 金额: $ 7.19万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10752101
• 关键词: Adult; Airway Disease; Asthma; Basal Cell; Binding; Biology; CRISPR interference; CRISPR-mediated transcriptional activation; Cell Line; Cells; Chronic Obstructive Pulmonary Disease; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Cystic Fibrosis; Data; Data Set; Development; Disease; Dryness; Environmental Risk Factor; Epithelial Cells; Epithelium; FGF10 gene; FGFR2 gene; Fibroblast Growth Factor; Gene Expression; Genes; Genetic; Genomics; Goals; Grant; Heterogeneity; Homeostasis; Human; Immune; Immunohistochemistry; In Situ; In Situ Hybridization; Ligands; Lung; Lung diseases; Malignant neoplasm of lung; Maps; Mesenchymal; Mesenchyme; Molecular; Morbidity - disease rate; Mus; Pathogenesis; Pathologic; Pathway interactions; Patients; Physicians; Play; Primary Ciliary Dyskinesias; Proliferating; Protocols documentation; Receptor Signaling; Recombinants; Regenerative Medicine; Resolution; Role; STEM field; Scientist; Signal Pathway; Signal Transduction; Structure of parenchyma of lung; System; Techniques; Technology; Testing; Tissue Sample; Trachea; airway remodeling; bronchial epithelium; career; cell type; differentiation protocol; directed differentiation; effective therapy; epithelium regeneration; induced pluripotent stem cell; knock-down; mortality; mouse model; non-genetic; novel; overexpression; programs; receptor; response; response to injury; self-renewal; single-cell RNA sequencing; stem cells; tool; transcriptomics

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.

项目总结/摘要 慢性阻塞性肺疾病（COPD）、哮喘、原发性纤毛运动障碍（PCD）和囊性纤维化 (CF)，代表由遗传和环境因素的复杂相互作用引起的气道疾病。他们是一个 发病率和死亡率的主要原因以及有效的治疗方法是可变的。哮喘的异质性和 COPD和罕见的PCD和CF对他们的研究提出了重大挑战。尽管他们的差异 这些气道疾病的发病机制共同具有协调反应， 上皮细胞、间充质细胞和免疫细胞，最终导致气道重塑[2]。这其中的根本 重塑是基底细胞（BC），气道的主要干细胞，以及它们的生态位[3]。遗传小鼠模型 表明由邻近间充质分泌Fgf配体与上皮Fgfr 2受体结合以维持BC [4][5]。Fgf与Hippo和Wnt 7 b在上皮-间充质信号传导中协同作用， 网络以实现上皮再生[6]-[8]。FGFR 2信号在人类BC中重要吗？令人惊奇的是， 尽管这一问题很重要，但所知甚少。我们将测试以下假设： 在维持成人气道基底细胞的自我更新能力中是必不可少的。在目标1中， 使用空间转录组学确定人气管和软骨间气道中的基底细胞小生境， 配体-受体信号传导分析和原位杂交技术。在目标2中，我们将测试FGFR 2 信号传导在使用人原代和诱导的人气道基底细胞维持人成人气道基底细胞中是必不可少的。 多能干细胞衍生的BC配备了基因编辑技术。在完成这一目标后，我们 已经确定了FGF在维持BC计划中的重要性。虽然这代表了一个雄心勃勃的 项目在短时间内，工具，细胞，协议，组织样本，并与适当的合作者 专业知识，都在。这个项目的范围和目标将使我接触到各种各样的技术 适合F32候选人。最终，这笔资金将产生令人兴奋的，新颖的数据， 作为一名研究肺部生物学的医生兼科学家，我将进入职业生涯的下一个阶段。