Diverse Homeostatic Roles for Distinct Macrophages in the Developing Lung Vasculature

不同巨噬细胞在发育中的肺血管系统中的多种稳态作用

基本信息

批准号：
10583456
负责人：
Cristina Maria Alvira
金额：
$ 62.08万
依托单位：
STANFORD UNIVERSITY
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：
2021-03-05 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10583456
关键词：
Acute Adult Air Sacs Alveolar Alveolar Macrophages Alveolus Anastomosis - action Area Basement membrane Birth Blood Vessels Bronchopulmonary Dysplasia Cell Communication Cells Chronic Complex Complication Coronary artery Cues Data Data Set Development Disease Distal Embryo Endothelial Cells Epithelial Cells Exhibits Gases Gene Expression Genes Genetic Transcription Growth Heart Heterogeneity Hyperoxia Image Immune Immunity Impairment In Situ In Situ Hybridization In Vitro Infant Injury Knockout Mice Life Ligands Location Lung Lung diseases Macrophage Mediating Molecular Myofibroblast Natural Immunity Natural regeneration Neuropeptides Organ Pathway interactions Pattern Phenotype Physiological Play Population Premature Birth Process Pulmonary Circulation Recovery Resolution Retina Role Serine Proteinase Inhibitors Signal Pathway Signal Transduction Smooth Muscle Myocytes Spatial Distribution Stromal Cell-Derived Factor 1 Surface Testing Tissues Transgenic Organisms Translating Vascular Permeabilities Vascular Smooth Muscle Vascular remodeling Vascularization angiogenesis arteriole blood vessel development chemokine conditional knockout genetic signature hindbrain in vivo lung development mouse model novel strategies postnatal postnatal development pulmonary function rapid growth receptor recruit repaired single-cell RNA sequencing therapeutic development transcriptomics

项目摘要

Alveolarization, the final stage of lung development occurring primarily postnatally, markedly increases gas exchange surface area. Rapid growth of the pulmonary vasculature by angiogenesis during early alveolarization drives distal lung growth, and disrupted angiogenesis impairs alveolarization. In other organs, specialized macrophages support angiogenesis by promoting blood vessel formation, providing survival and migratory cues to EC, and facilitating vascular anastomoses. However, the role of macrophages in the developing pulmonary vasculature remains entirely unknown. We recently embarked on a project employing single cell RNA-sequencing to define macrophage diversity during late embryonic and early postnatal lung development. Macrophages are extremely heterogenous with diverse phenotypes that are lineage- and tissue- specific, and highly influenced by the microenvironment. Preliminary data in this proposal demonstrate a tremendous increase in macrophage diversity after birth. Specialized, highly proliferative macrophages present before birth are replaced after birth by a complex and dynamic mixture of diverse macrophage subtypes exhibiting unique gene signatures, developmental gradients in gene expression, and specific locations within the lung suggesting distinct functions in tissue remodeling, angiogenesis, and immunity. Interestingly, a subset of embryonic macrophages was found to completely encircle small arterioles and express numerous genes that regulate lung branching, angiogenesis, and EC phenotype. After birth, these cells transitioned to an intermediate subset present only during the first few weeks of postnatal life that expressed additional tissue remodeling genes. Taken together, our data suggest the hypothesis that distinct macrophage populations support alveolarization by regulating pulmonary vascular development through the expression of factors that influence vascular growth and remodeling, which will be tested through three specific aims. Aim 1 will combine multiplexed in situ hybridization, lineage tracing, studies in primary EC and macrophages, and advanced imaging in transgenic and knock-out mice to define the role of specific macrophage subsets in modulating EC phenotype and regulating lung parenchymal and vascular growth. Aim 2 will utilize multiplexed in situ hybridization, conditional knock out mouse models, and ligand-receptor profiling of single cell datasets from pulmonary EC and macrophages to probe pathways mediating macrophage-EC communication. Finally, Aim 3 will determine if chronic hyperoxia alters diversity and phenotype of the lung macrophages during acute injury and after recovery, and specifically impairs developmental and homeostatic functions of lung macrophages. The successful completion of these studies will provide a multifaceted view of the diverse functions of lung macrophages during embryonic and early postnatal development at single cell resolution, and identify new pathways that could be directly translated into novel strategies to modulate vascular growth and regeneration in diseases marked by impaired pulmonary angiogenesis.

牙槽化，肺发育的最后阶段主要发生，明显增加气体交换表面积。早期血管生成肺脉管系统的快速生长肺泡化驱动肺部远端生长，并破坏血管生成会损害肺泡化。在其他器官中专门的巨噬细胞通过促进血管形成，提供生存和 EC的迁移提示，并促进血管吻合。但是，巨噬细胞在发育的肺脉管系统仍然完全未知。我们最近着手采用一个项目单细胞RNA - 在胚胎晚期和早期肺中定义巨噬细胞的多样性发展。巨噬细胞是极度异质的，具有多种表型，这些表型是谱系和组织的特定于微环境的高度影响。该提案中的初步数据证明了出生后巨噬细胞多样性的巨大增加。存在专门的，高度增殖的巨噬细胞出生前出生前通过各种巨噬细胞的复杂而动态的混合物代替出生前表现出独特的基因特征，基因表达中的发育梯度以及特定位置肺在组织重塑，血管生成和免疫力中表现出不同的功能。有趣的是，一个子集发现胚胎巨噬细胞完全包围小动脉并表达许多基因调节肺分支，血管生成和EC表型。出生后，这些细胞过渡到中间体子集仅在产后生命的前几周出现，该寿命表达了其他组织重塑基因。综上所述，我们的数据表明，不同的巨噬细胞种群支持牙槽化通过调节肺血管发育，表达影响血管生长的因素和重塑，将通过三个特定目标进行测试。 AIM 1将结合原位多路复用杂交，谱系跟踪，一级EC和巨噬细胞的研究以及转基因和转基因的高级成像敲除小鼠，以定义特定巨噬细胞亚群在调节EC表型和调节肺的作用实质和血管生长。 AIM 2将利用多路复用的原位杂交，有条件的敲除鼠标从肺EC和巨噬细胞到探测途径的单细胞数据集的模型以及配体受体分析介导巨噬细胞-EC通信。最后，AIM 3将确定慢性高氧是否改变了多样性和急性损伤和恢复后肺巨噬细胞的表型，特别会损害肺巨噬细胞的发育和稳态功能。这些研究的成功完成将在胚胎和早期产后，提供多方面的肺巨噬细胞功能的多方面视图在单细胞分辨率下开发，并确定可以直接转化为新颖的新途径调节肺部受损标志的疾病中血管生长和再生的策略血管生成。