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Characterization of a New Epithelial Cell in the Developing Human Lung

发育中的人肺中新上皮细胞的表征

基本信息

批准号：
10313576
负责人：
Ansley Corinne Semack Conchola
金额：
$ 4.06万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10313576
关键词：
Antibodies Archives Bar Codes Blocking Antibodies Bronchi Cell Differentiation process Cell Maintenance Cell Surface Proteins Cell physiology Cell surface Cells Complex Computer software Cues Cystic Fibrosis Transmembrane Conductance Regulator Data Decision Making Development Distal Dominant-Negative Mutation ES01 Epithelial Epithelial Cells Fetal Lung Fetal Tissues Fluorescence-Activated Cell Sorting Gene Expression Gene Expression Profile Goals Human In Situ Light Lung Methods Modeling Morphogenesis Mus Organoids Pharmacology Population Process Publishing Reagent Regulation Role Route Secretory Cell Signal Transduction Stains Structure of parenchyma of lung Testing Time Tissue Banks Trachea airway epithelium base cell type cohort data repository experimental study fetal gain of function human embryonic stem cell in vitro Model in vivo induced pluripotent stem cell inhibitor/antagonist loss of function lung development novel progenitor single-cell RNA sequencing spatiotemporal

项目摘要

PROJECT ABSTRACT The lung buds undergo a process known as branching morphogenesis, which establishes the complex, arborized network of the lung1–7. During branching, a specialized population of lung progenitors at the tips of the branches, bud tip progenitors (BTPs), give rise to all epithelial cell types of the lung including cells of the proximal airway (trachea, bronchi) and cells of the distal lung (alveoli)1,8–16. We recently performed single-cell RNA sequencing (scRNAseq) on the human fetal lung from 10 to 21 weeks gestation16, and identified a novel epithelial cell population characterized by a unique gene expression profile: SCGB3A2HI/SFTPB HI/CFTRHI which we refer to as Fetal Airway Secretory (FAS) cells. We have shown that human BTPs grown in culture can give rise to FAS cells, and they have been verified in the small airways in situ. My preliminary data suggest that FAS cells are transient, dwindling in number across development. Further, our data suggests that FAS cells are uniquely human, and do not exist in mice. Since nothing is known about this newly identified population, the goal of this proposal is to interrogate the function, differentiation potential, and regulation of FAS cells. My preliminary data suggests that: (a) FAS cells may be a developmental precursor state that exists while BTPs are en route to becoming proximal airway and (b) FAS cell development is regulated by NOTCH signaling, a critical regulator of cell fate decision making in the developing lung17,18. Based on my preliminary data, I hypothesize that FAS cells are a transitional progenitor population that give rise to all airway cell types and NOTCH signaling regulates the transition from BTP-to-FAS to committed airway cell types. Using a novel cell barcoding lineage tracing approach in a human lung organoid model, I aim to 1) test the function of FAS cells, specifically if they differentiate into proximal airway cells. I also aim to 2) investigate the role of NOTCH signaling in FAS cells using gain-of-function and loss-of-function lentiviral constructs, NOTCH blocking antibodies, and pharmacologic inhibitors that activate or inhibit NOTCH signaling in an organoid model.

项目摘要肺芽经历一个称为分支形态发生的过程，该过程建立了复杂的树状结构肺网络1-7。在分支过程中，分支尖端有一群特殊的肺祖细胞，芽尖祖细胞 (BTP) 产生肺的所有上皮细胞类型，包括近端气道细胞（气管、支气管）和远端肺细胞（肺泡）1,8-16。我们最近对 10 至 21 周的人类胎儿肺部进行了单细胞 RNA 测序 (scRNAseq) gestation16，并鉴定出一种新型上皮细胞群，其特征在于独特的基因表达谱： SCGB3A2HI/SFTPB HI/CFTRHI 我们称之为胎儿气道分泌 (FAS) 细胞。我们已经证明在培养物中生长的人类 BTP 可以产生 FAS 细胞，并且它们已经在小气道中得到了原位验证。我的初步数据表明，FAS 细胞是短暂的，在发育过程中数量不断减少。此外，我们的数据表明，FAS 细胞是人类独有的，在小鼠体内不存在。由于对此一无所知新识别的群体，该提案的目标是询问功能、分化 FAS 细胞的潜力和调节。我的初步数据表明：(a) FAS 细胞可能是 BTP 存在时存在的发育前体状态正在成为近端气道，并且 (b) FAS 细胞发育受 NOTCH 信号传导调节，NOTCH 信号传导是一种发育中的肺细胞命运决策的关键调节因子17,18。根据我的初步数据，我假设 FAS 细胞是一种过渡祖细胞群，可产生所有气道细胞类型，并且 NOTCH 信号传导调节从 BTP 到 FAS 到定型气道细胞类型的转变。使用新型细胞在人肺类器官模型中采用条形码谱系追踪方法，我的目标是 1) 测试 FAS 细胞的功能，特别是如果它们分化为近端气道细胞。我还致力于 2) 研究 NOTCH 信号传导的作用在 FAS 细胞中使用功能获得和功能丧失的慢病毒构建体、NOTCH 阻断抗体和在类器官模型中激活或抑制 NOTCH 信号传导的药物抑制剂。