Investigating the role of mesenchymal R-Spondin2 during human lung development

研究间充质 R-Spondin2 在人肺发育过程中的作用

基本信息

批准号：
10453627
负责人：
Renee Frances Conway
金额：
$ 1.9万
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：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2022-11-11
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10453627
关键词：
Adenoviruses Agonist Air Algorithms Behavior Binding Biological Models Cell membrane Cell physiology Cells Cessation of life Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Complex Data Defect Development Distal Endoderm Epithelial Epithelial Cells Event Exons Fetal Lung Fluorescent in Situ Hybridization Genes Goals Growth Heterogeneity Home Human Human Development Immunofluorescence Immunologic In Vitro Knowledge LGR5 gene Lead Ligands Liquid substance Longitudinal Studies Lung Maintenance Maps Mesenchymal Mesenchyme Methods Morphogenesis Mus Mutation Organoids Population Pregnancy Primitive foregut structure Process Proliferating Recombinants Regulation Respiratory Failure Respiratory System Role Sampling Signal Transduction Smooth Muscle Myocytes Specific qualifier value Stains Structure of parenchyma of lung Supporting Cell System Systems Development Testing Tube WNT Signaling Pathway WNT2 gene Work beta catenin cell type epithelial stem cell experimental study fetal human data human fetal lung tissue human pluripotent stem cell in vitro Model in vivo lung development premature progenitor receptor single-cell RNA sequencing small molecule stem cells transcriptome vpr Genes

项目摘要

PROJECT ABSTRACT Mammalian respiratory system development is regulated by complex reciprocal signaling events that take place between the epithelial cells lining the airways and the surrounding mesenchymal cells. Despite the lung mesenchyme being home to cell types that provide crucial signals during development, the identity and function of specific cell types in the lung mesenchyme are poorly defined, especially in the context of human development. To interrogate mesenchymal cell heterogeneity in the developing human lung, our lab has conducted single cell RNA sequencing on multiple human lung samples spanning 8-22 weeks of gestation. Using cell-clustering approaches, we identified a mesenchymal cell population highly enriched for expression of the WNT agonist R-Spondin2 (RSPO2). Strikingly, mutations in this gene cause severe lung aplasia in humans. My co-fluorescent in situ hybridization (FISH) and immunofluorescence (IF) data from human lung tissue sections show that RSPO2 is expressed adjacent to proliferating distal epithelial progenitor cells and that the RSPO2 receptor, LGR5, is expressed almost exclusively in the epithelial progenitor cells. These data suggest that RSPO2+ mesenchymal cells may have a critical role in regulating epithelial progenitor cells during human lung development. The goals of this proposal are to: 1) interrogate mesenchymal cell heterogeneity and function in the context of epithelial progenitor cell regulation in the developing human lung and 2) explain a functional role for RSPO2+ mesenchymal cells during human lung development.

项目摘要哺乳动物呼吸系统开发受到复杂的相互信号事件的调节放置在衬在气道和周围间充质细胞的上皮细胞之间。尽管有肺间充质是在开发过程中提供关键信号的细胞类型的故乡肺间充质中特定细胞类型的功能的定义很差，尤其是在人类的背景下发展。为了询问发育中的人肺中间充质细胞异质性，我们的实验室有在妊娠8-22周的多个人肺样品上进行了单细胞RNA测序。使用细胞聚类方法，我们确定了一个高度富集表达的间充质细胞群 Wnt激动剂R-Spondin2（RSPO2）令人惊讶的是，该基因中的突变引起严重的肺发育不全人类。我的共荧光原位杂交（FISH）和免疫荧光（如果）来自人类肺的数据组织切片表明RSPO2表达与增殖的远端上皮祖细胞和 RSPO2受体LGR5几乎完全在上皮祖细胞中表达。这些数据表明RSPO2+间充质细胞在调节上皮祖细胞中可能具有关键作用人肺发育。该提议的目标是：1）询问间充质细胞异质性和在发展中肺的上皮祖细胞调节的背景下的功能，2）解释人肺发育过程中RSPO2+间充质细胞的功能作用。