Functional interrogation of a novel SCGB3A2+/SFTPB+ cell in the human airway

人类气道中新型 SCGB3A2 /SFTPB 细胞的功能询问

• 批准号: 10555804
• 负责人: Jason Spence
• 金额: $ 58.22万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10555804
• 关键词: ASCL1 gene; Atlases; Bar Codes; Basal Cell; Biology; Cell Differentiation process; Cell Lineage; Cell surface; Cells; Conceptions; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Epithelial Cells; Epithelium; Gene Expression Profile; Genes; Goals; Human; In Vitro; Lung; MADH2 gene; Mediating; Methods; Modeling; Mus; Neuroendocrine Cell; Organoids; Phase; Population; Publishing; Reporting; Secretory Cell; Signal Transduction; System; Testing; Time Series Analysis; Tissues; cell type; experimental study; fetal; follow-up; genetic signature; in vitro Model; inhibitor; insight; novel; pharmacologic; progenitor; single-cell RNA sequencing; stem cells; tool

ABSTRACT During a recent single cell RNA sequencing analysis of the developing human lung, we identified a cell population characterized by a unique gene expression profile that has not previously been reported in the human or mouse lung. These cells express SCGB3A2/SFTPB/CFTR, but lack markers for other well characterized cell types such as club cells (SCGB1A1) or ionocytes (FOXI1). We refer to these as Fetal Airway Secretory (FAS) cells given the expression of several secretory genes within the transcriptional signature. Since nothing is known about FAS cells, and there is no analogous population in the murine lung, the overarching goal of this proposal is to both describe these cells in detail, to interrogate their differentiation potential, and to determine how these cells are regulated. By using an in vitro model of the developing human lung epithelium called human Bud Tip Progenitor (BTP)-organoids, preliminary data shows that BTP-organoids can give rise to both TP63+ progenitors and FAS cells during airway differentiation, and that this differentiation can be modulated by TGFb/BMP/SMAD signaling. Single cell barcode-based lineage tracing during the transition from BTP-to-airway suggest that clones of FAS cells or basal cells give rise to distinct subsets of airway cells, with FAS cells giving rise to pulmonary neuroendocrine cells (PNECs) and a subset of C6+ multiciliated (MC) cells, while TP63+ progenitor cells give rise to secretory cells and MUC16+ MC cells. Based on this data, we will test the overarching hypothesis that FAS cells differentiate distinct airway progenitor cell that give rise to a subset of airway cell types. Given the novelty of this cell type, and the unique tools that we have developed to assess the differentiating human epithelium in vitro, understanding this cell population will provide foundational descriptive and mechanistic insights into human lung biology.

抽象的 在最近对发育中的人肺进行的单细胞 RNA 测序分析中，我们发现了一个细胞 以独特的基因表达谱为特征的人群，该基因表达谱以前未在文献中报道过 人或小鼠的肺。这些细胞表达 SCGB3A2/SFTPB/CFTR，但缺乏其他孔的标记 表征细胞类型，例如俱乐部细胞 (SCGB1A1) 或离子细胞 (FOXI1)。我们将这些称为胎儿气道 分泌（FAS）细胞在转录特征中表达多个分泌基因。 由于对 FAS 细胞一无所知，并且小鼠肺中没有类似的细胞群，因此 该提案的总体目标是详细描述这些细胞，询问它们的分化 潜力，并确定这些细胞是如何受到调节的。通过使用人类发育的体外模型 肺上皮称为人芽尖祖细胞（BTP）-类器官，初步数据表明BTP-类器官 在气道分化过程中可以产生 TP63+ 祖细胞和 FAS 细胞，并且这种分化 可以通过 TGFb/BMP/SMAD 信号传导进行调节。基于单细胞条形码的谱系追踪 从 BTP 到气道的转变表明 FAS 细胞或基底细胞的克隆产生了不同的亚群 气道细胞，其中 FAS 细胞产生肺神经内分泌细胞 (PNEC) 和 C6+ 的子集 多纤毛 (MC) 细胞，而 TP63+ 祖细胞产生分泌细胞和 MUC16+ MC 细胞。基于 根据这些数据，我们将检验 FAS 细胞分化不同气道祖细胞的总体假设 产生气道细胞类型的子集。鉴于这种细胞类型的新颖性以及我们所使用的独特工具 已开发出体外评估分化的人类上皮细胞，了解该细胞群将有助于 为人类肺生物学提供基本的描述性和机制性见解。