Lung transplant injury drives chronic lung allograft dysfunction via recruitment ofmonocyte-derived alveolar macrophages

肺移植损伤通过单核细胞衍生的肺泡巨噬细胞的募集导致慢性肺同种异体移植功能障碍

• 批准号: 10682438
• 负责人: Alexander Misharin
• 金额: $ 73.76万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10682438
• 关键词: Acute; Allografting; Alveolar Macrophages; Automobile Driving; Biological Markers; Biopsy; Bronchiolitis Obliterans; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; CSF1 gene; CSF1R gene; Caring; Cells; Chronic; Clinical; Colony-Stimulating Factor Receptors; Computer Analysis; Cre lox recombination system; Data; Data Set; Development; Disease; Endothelium; Event; Experimental Genetics; Extravasation; Fibroblasts; Fibrosis; Functional disorder; Genetic; Heart-Lung Transplantation; Human; Immune; In Situ; Injury; Life; Link; Liquid substance; Lung; Lung Transplantation; Lung diseases; Macrophage Colony-Stimulating Factor; Maintenance; Methods; Molecular; Morbidity - disease rate; Mus; Neutrophil Infiltration; Operative Surgical Procedures; Pathogenicity; Pathologic; Pathology; Pathway interactions; Patients; Perioperative; Play; Population; Procedures; Proliferating; Publishing; Pulmonary Fibrosis; RNA; Reperfusion Injury; Reporting; Residencies; Role; Sampling; Semaphorins; Signal Transduction; Solid; Spleen; Structure of parenchyma of lung; Syndrome; Testing; Therapeutic; Tissues; Transplant Recipients; Transplant Surgeon; Transplantation; Trauma; Work; autocrine; biomarker identification; cell type; clinical phenotype; effective therapy; epithelial injury; fibrotic lung disease; graft dysfunction; human data; improved; inducible Cre; lung allograft; lung injury; monocyte; mortality; mouse model; new therapeutic target; organ transplant recipient; plexin; post-transplant; prevent; programs; recruit; response; retransplantation; single cell analysis; single-cell RNA sequencing; transcriptome sequencing

Project Summary: For many patients with advanced lung disease, lung transplantation remains the only viable therapeutic option to extend life. Yet the 5-year survival for lung transplant patients is 54%, the worst among solid organ transplant recipients. Chronic lung allograft dysfunction (CLAD) is the leading cause of morbidity and late mortality after lung or heart-lung transplantation. Hence, identification of biomarkers and novel therapeutic targets is essential to prevent or treat CLAD and extend survival after lung transplantation. While multiple factors contribute to CLAD, early events after the lung transplant procedure play a crucial role in setting the stage for subsequent CLAD. These include surgical trauma and ischemia-reperfusion injury that activates circulating and resident immune cells followed by the endothelial injury and immune cell extravasation. We have reported that monocytes, recruited to the injured lung, can establish a long-term residency and differentiate into pathogenic monocyte-derived alveolar macrophages. We have causally linked monocyte-derived alveolar macrophages to tissue-remodeling and fibrosis (resembling CLAD) using a genetic deletion strategy. Moreover, using unbiased single-cell transcriptomic profiling (RNA-seq) of explanted lung tissue from the patients with pulmonary fibrosis and biopsies of the donor lung (both obtained during lung transplantation by lung transplant surgeon – Ankit Bharat, key contributor to this proposal), we identified a distinct population of pathogenic alveolar macrophages exclusively present in patients with pulmonary fibrosis. Our computational analyses of single-cell RNA-seq data suggest that monocyte-derived alveolar macrophages are guided to their new pathogenic niches via plexin D1/signaling and are uniquely maintained by M-CSF/M- CSFR signaling. Consistent with this hypothesis, targeting M-CSF/M-CSFR signaling specifically eliminated monocyte-derived alveolar macrophages and ameliorated pathology. We present preliminary data from mouse models and patients with chronic lung allograft dysfunction supporting relevance of this mechanism for CLAD. We will thus use mouse models and samples from lung transplant patients to test the hypothesis that pathogenic monocyte-derived alveolar macrophages, recruited during the initial peri-transplant injury, establish long term residency via plexin D1/semaphorin signaling and are maintained via M-CSF/M- CSFR signaling to drive CLAD in three interrelated aims: Aim 1: To determine whether monocyte-derived alveolar macrophages recruited to the transplanted lung within day of the transplant are maintained by M-CSF/M-CSFR signaling. Aim 2: To determine whether pathogenic monocyte-derived alveolar macrophages are localized to regions of lung fibrosis lung via plexin D1/semaphorin signaling. Aim 3: To determine whether the emergence of aberrant alveolar macrophages with increased expression of PLXND1 and autocrine M-CSF/M-CSFR signaling can be identified in BAL fluid from patients with early CLAD.

项目摘要：对于许多患有晚期肺部疾病的患者，肺移植仍然是唯一的 可行的疗法延长寿命的选择。然而，肺移植患者的5年生存率为54％，是最糟糕的 在固体器官移植受体中。慢性肺同种异体功能障碍（CLAD）是 肺或心肺移植后发病和晚期死亡率。因此，识别生物标志物和 新型的热目标对于预防或治疗肺移植后延伸和延长生存至关重要。 虽然多种因素导致外壳，但肺移植手术后的早期事件起着至关重要的作用 在为随后的外壳设置阶段的角色。这些包括手术创伤和缺血 - 重新灌注 激活循环的损伤，居民免疫接种，然后是内皮损伤和免疫细胞细胞 奢侈。我们报告说，被招募到受伤的肺部的单核细胞可以建立长期 分离并分化为致病的单核细胞衍生的肺泡巨噬细胞。我们有因果关系 单核细胞衍生的肺泡巨噬细胞使用仿制药 删除策略。此外，使用外植物肺的无偏单细胞转录组分析（RNA-seq） 来自肺纤维化和供体肺活检患者的组织（均在肺部获得 肺移植外科医生的移植 - Ankit Bharat，该提案的主要贡献者），我们确定了一个 肺纤维化患者中仅存在的致病性肺泡巨噬细胞的不同种群。 我们对单细胞RNA-seq数据的计算分析表明，单核细胞衍生的肺泡巨噬细胞 通过PLEXIN D1/信号传导将其引导到其新的致病壁细分市场，并由M-CSF/M-独特地维护 CSFR信号传导。与该假设一致，针对M-CSF/M-CSFR信号传导专门消除 单核细胞衍生的肺泡巨噬细胞和改善病理学。我们介绍鼠标的初步数据 模型和患有慢性肺同类功能障碍的患者支持该机制与外壳的相关性。 因此，我们将使用小鼠模型和来自肺移植患者的样本来检验以下假设。 致病性单核细胞衍生的肺泡巨噬细胞，在最初的移植物损伤期间募集 通过PLEXIN D1/Semaphorin信号传导建立长期住所，并通过M-CSF/M-维持 CSFR信号传导以三个相互关联的目的驱动外壳： 目标1：确定单核细胞衍生的肺泡巨噬细胞是否募集到移植的肺部 移植日通过M-CSF/M-CSFR信号传导维持。 目标2：确定病原单核细胞衍生的肺泡巨噬细胞是否定位于 肺纤维化肺通过PLEXIN D1/Semaphorin信号传导。 目标3：确定异常肺泡巨噬细胞的出现是否增加 PLXND1和Autocrine M-CSF/M-CSFR信号可以在早期外壳患者的BAL液中鉴定出来。