Macrophage Modulation of Lung Fibrosis

肺纤维化的巨噬细胞调节

• 批准号: 9264201
• 负责人: Harris R Perlman
• 金额: $ 66.72万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9264201
• 关键词: Adenoviruses; Adoptive Transfer; Alpha Cell; Alveolar Macrophages; Apoptotic; Area; Aspartic Acid; Attenuated; Biological Models; Bleomycin; CASP8 gene; Cause of Death; Cell Death; Cells; Cessation of life; Complex; Cysteine; Data; Development; Disease; Fibrosis; Flow Cytometry; Gene Expression; Genes; Genetic Transcription; Heterogeneity; Homologous Gene; Human; Human body; Knockout Mice; Lung; Lung Transplantation; Mediating; Modeling; Molecular; Molecular Profiling; Mus; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Peptide Hydrolases; Phosphotransferases; Play; Population; Population Heterogeneity; Pulmonary Fibrosis; RIPK1 gene; RIPK3 gene; Recruitment Activity; Research Personnel; Role; Scleroderma; Severities; Signal Transduction; Standardization; Systemic Scleroderma; Testing; Time; Tissues; Transforming Growth Factor beta; Work; course development; genetic signature; human disease; humanized mouse; lung development; lung injury; macrophage; monocyte; mouse model; mutant; novel; peripheral blood; programs; receptor; reconstitution; response; sialic acid binding Ig-like lectin; tool; transcriptome sequencing

Pulmonary fibrosis has emerged as the leading cause of death in patients with systemic sclerosis (SSc)/scleroderma, yet currently available therapies are only marginally effective. While recent work from several groups of investigators suggests a key role for lung macrophages in the development of pulmonary fibrosis in patients with SSc, this remains an area of controversy as the specific population(s) of lung macrophages that drive fibrosis and the molecular mechanisms by which they do so remain largely unknown. In advance of this proposal, we have developed novel tools to elucidate macrophage heterogeneity in the mouse and human lung including lineage tagging, flow cytometry and transcriptional profiling in order to examine the differential role of tissue-resident alveolar macrophages, as compared to the monocyte-derived alveolar macrophages recruited to the lung, in the pathogenesis of fibrosis. Using unbiased transcriptional analysis (RNAseq) of flow-sorted macrophage populations over the course of the development of experimental fibrosis, we identified genetic signatures of monocyte to alveolar macrophage differentiation and fibrosis. Moreover, we have developed a murine model, which lacks the pathogenic monocyte-derived macrophage and is thus unable to develop lung fibrosis. Specifically, mice with macrophage-specific deletion of caspase-8, a cysteine-aspartic acid protease originally identified as a key initiator of the apoptotic death receptor pathway and suppressor of necroptosis (CreLysMCasp8fl/fl or CreCD11cCasp8fl/fl), showed significantly attenuated fibrosis and an inability for recruited monocytes to differentiate into pro-fibrotic monocyte-derived Siglec Flow macrophages following intratracheal treatment with either bleomycin or an adenovirus encoding an active form of TGF-β as compared to Casp8fl/fl, CreLysM or CreCD11c mice. Thus, this model is an ideal to tool to understand the differentiation of monocytes into pro-fibrotic alveolar macrophages in response to lung injury. These model systems will allow us to apply the transcriptional data to the study of monocytes from patients with SSc differentiated into alveolar macrophages in the lungs of humanized mice and alveolar macrophages obtained from patients with SSc at the time of lung transplantation. Our studies will test the hypothesis that monocyte differentiation into pro-fibrotic Siglec Flow alveolar macrophages is essential for the development of lung fibrosis in both mice and humans in three interrelated specific aims.

肺纤维化已成为系统性硬化症患者的主要死亡原因 (SSc)/硬皮病，但目前可用的治疗方法仅有轻微效果。虽然最近的工作来自 几组研究人员认为肺巨噬细胞在肺间质瘤的发生发展中起关键作用 SSc患者的纤维化，这仍然是一个有争议的领域，因为肺的特定人群(S) 推动纤维化的巨噬细胞及其分子机制在很大程度上仍然存在 未知。在这项提议之前，我们已经开发了新的工具来阐明巨噬细胞的异质性。 包括谱系标记、流式细胞术和转录图谱，以便在小鼠和人类肺中 与单核细胞来源的肺泡巨噬细胞相比，观察组织内驻留的肺泡巨噬细胞的不同作用 肺泡巨噬细胞募集到肺内，参与纤维化的发病机制。使用不偏不倚的转录 实验发展过程中流动分选巨噬细胞种群的分析(RNAseq) 纤维化，我们确定了单核细胞到肺泡巨噬细胞分化和纤维化的遗传特征。 此外，我们还开发了一种小鼠模型，该模型缺乏致病的单核细胞来源的巨噬细胞和 因此不能发展成肺纤维化。具体地说，巨噬细胞特异性缺失caspase-8，a的小鼠 半胱氨酸-天冬氨酸蛋白酶最初被认为是凋亡死亡受体途径的关键启动者 和坏死性下垂的抑制物(CreLysMCasp8fl/fl或CreCD11cCasp8fl/fl)显示显著减轻纤维化 重新招募的单核细胞不能分化为促纤维化的单核细胞来源的Siglec流 气管内应用博莱霉素或编码活性形式的腺病毒治疗后的巨噬细胞 与Casp8fl/fl、CreLysM和CreCD11c小鼠比较，结果表明，Casp8fl/fl、CreLysM和CreCD11c小鼠的β基因表达水平均高于Casp8fl/fl、CreLysM和CreCD11c小鼠。因此，该模型是理解该模型的理想工具 肺损伤后单核细胞向促纤维化的肺泡巨噬细胞分化。这些模型 系统将允许我们将转录数据应用于对SSc患者单核细胞的研究 人源化小鼠肺内分化为肺泡巨噬细胞并获得肺泡巨噬细胞 来自肺移植时患有SSc的患者。我们的研究将检验这一假设 单核细胞分化为促纤维化的Siglec Flow肺泡巨噬细胞对 在三个相互关联的特定目标中发展小鼠和人类的肺纤维化。