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.

项目概要：对于许多晚期肺病患者来说，肺移植仍然是唯一的选择 然而，肺移植患者的 5 年生存率为 54%，是最差的。 在实体器官移植受者中，慢性肺同种异体移植功能障碍（CLAD）是导致该病的主要原因。 因此，生物标志物的鉴定和肺或心肺移植后的发病率和晚期死亡率。 新的治疗靶点对于预防或治疗 CLAD 以及延长肺移植后的生存至关重要。 虽然导致 CLAD 的因素有多种，但肺移植手术后的早期事件起着至关重要的作用。 为随后的 CLAD 奠定基础，其中包括手术创伤和缺血再灌注。 激活循环和驻留免疫细胞的损伤，然后是内皮损伤和免疫细胞 我们已经报道，募集到受伤肺部的单核细胞可以建立长期的细胞外渗。 驻留和分化为致病性单核细胞来源的肺泡巨噬细胞之间存在因果关系。 单核细胞来源的肺泡巨噬细胞利用基因进行组织重塑和纤维化（类似于 CLAD） 此外，使用外植肺的无偏单细胞转录组分析（RNA-seq）。 肺纤维化患者的组织和供体肺的活检（均在肺纤维化过程中获得） 由肺移植外科医生（Ankit Bharat，该提案的主要贡献者）进行移植），我们确定了 独特的致病性肺泡巨噬细胞群仅存在于肺纤维化患者中。 我们对单细胞 RNA-seq 数据的计算分析表明，单核细胞来源的肺泡巨噬细胞 通过丛蛋白 D1/信号传导引导至新的致病位，并由 M-CSF/M- 独特维持 与该假设一致的是，针对 M-CSF/M-CSFR 信号传导被特异性消除。 我们提供了来自小鼠的初步数据。 患有慢性肺同种异体移植功能障碍的模型和患者支持这种机制与 CLAD 的相关性。 因此，我们将使用小鼠模型和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，在最初的围移植损伤期间招募， 通过 plexin D1/semaphorin 信号传导建立长期驻留，并通过 M-CSF/M- 维持 CSFR 信号驱动 CLAD 实现三个相互关联的目标： 目标 1：确定单核细胞来源的肺泡巨噬细胞是否在 移植日由 M-CSF/M-CSFR 信号维持。 目标 2：确定致病性单核细胞来源的肺泡巨噬细胞是否定位于 通过丛蛋白 D1/信号蛋白信号传导导致肺纤维化。 目标 3：确定是否出现异常肺泡巨噬细胞，其表达增加 可以在早期 CLAD 患者的 BAL 液中鉴定出 PLXND1 和自分泌 M-CSF/M-CSFR 信号。