Airway epithelial cell and lymphocyte interactions in chronic lung allograft dysfunction pathogenesis

慢性肺同种异体移植功能障碍发病机制中气道上皮细胞和淋巴细胞的相互作用

基本信息

批准号：
10684231
负责人：
JOHN GREENLAND
金额：
$ 61.44万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-15 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10684231
关键词：
Activated Lymphocyte Address Affect Airway Fibrosis Alloantigen Anatomy Apoptosis Apoptotic Automobile Driving Bioinformatics Biometry Bronchoalveolar Lavage Bronchoalveolar Lavage Fluid Brush Cell CD4 Positive T Lymphocytes CD8B1 gene Cell Culture System Cell Culture Techniques Cell Death Cell Hypoxia Cell surface Cells Chronic Cicatrix Circulation Clinical Trials Cohort Studies Complement Cytolysis Cytometry Cytotoxic T-Lymphocytes Data Detection Development Diagnosis Disease Distal Epithelial Cells Exposure to Fibrosis Flow Cytometry Functional disorder Gene Expression Gene Expression Profile Genes Genetic Transcription Heat shock proteins Hypoxia Hypoxia Pathway Image Immune Immune response Immunity Immunobiology In Vitro Inflammation Infrastructure Institution Investigation Kinetics Ligands Light Link Lung Lung Transplantation Lung diseases Lymphocyte Lymphoid Cell Macrophage Mediating Methods OX40 Pathogenesis Pathologic Pathology Pathway Analysis Pathway interactions Phenotype Play Prevention Primary Cell Cultures Proteins Publishing Quality of life Reporting Role Signal Transduction Source Stimulus Stress Structure of parenchyma of lung Surrogate Markers T cell response T-Lymphocyte T-cell inflamed TNF gene TNF-related apoptosis-inducing ligand TNFRSF10A gene TNFRSF5 gene TNFRSF8 gene TNFSF5 gene Techniques Testing Time Transcript Transplant Recipients Transplantation Up-Regulation Validation Vascular blood supply airway epithelium apoptosis in lymphocytes cell type chemokine clinical phenotype clinical subtypes cohort cytokine cytotoxic effector T cell fibrotic lung functional improvement improved innovation insight lung allograft member multidisciplinary novel novel marker novel strategies post-transplant prevent progenitor programs pulmonary function recruit respiratory hypoxia response segregation single-cell RNA sequencing stem cells success synergism targeted therapy trials tool transcriptome transcriptome sequencing translational research program transplant centers vascular inflammation

项目摘要

The major barrier to long term survival following lung transplantation is a progressive loss of lung function, termed chronic lung allograft dysfunction (CLAD), for which constrictive fibrosis in small airways is a pathologic hallmark. CLAD affects over half of lung transplant recipients by 4 years post-transplant and negates much of the quality of life and functional improvements associated with transplantation. Pitt, Toronto, and UCSF Lung Transplant programs have refined transcriptional analysis of small airway brushings from lung transplant recipients as a novel technique to understand the gene expression changes at the anatomical site where CLAD pathology develops. We have published gene expression changes associated with CLAD validated across our centers. This proposal leverages this innovative approach to understand mechanisms of CLAD pathogenesis. Our preliminary data show an early upregulation of hypoxia pathways in airway brushings including genes that recruit and activate cytotoxic T lymphocytes using both airway epithelial cell culture in hypoxic conditions and pathway analysis of airway brush transcriptomes. This hypoxia signaling may reflect disordered microvasculature, absent bronchial circulation, and vascular inflammation associated with lung transplant. In recruited T lymphocytes, we also observed upregulation of tumor necrosis factor superfamily (TNFSF) genes, which are major drivers of apoptosis in lymphocyte targets. Our data show preferential apoptosis in airway club cells, the protectors and progenitors of small airways, in association with upregulated TNF-related apoptosis- inducing ligand (TRAIL) expression. Our single cell investigations in airway brushings and bronchoalveolar lavage (BAL) fluid show the segregation of these pathways across epithelial and lymphoid cell types. Based on these data, we hypothesize that airway hypoxia precedes TNFSF gene expression and T cell-mediated airway club cell apoptosis that drive CLAD pathogenesis. To test this hypothesis, we will generate parallel cohorts investigating bulk and single cell transcriptomes of CLAD versus controls across three centers, allowing for rigorous cross-validation of gene expression signatures. We will complement these studies with cell culture- based methods to determine mechanisms driving these gene expression changes. In Aim 1, we will quantify hypoxia-related transcripts in airway brush cells with respect to CLAD and determine how hypoxia can promote lymphocytic inflammation. In Aim 2, we will determine the cellular sources and kinetics of TNFSF co-stimulatory molecule expression in CLAD using airway brushes and BAL fluid. In Aim 3, we will investigate whether TRAIL preferentially induces club cell apoptosis. The synergy of three large lung-transplant translational research programs with world-class cross-institutional biostatistical infrastructures provides a unique opportunity to address this hypothesis rigorously. The cell-specific gene expression signatures over the time course of CLAD development that will be generated through this study are critically needed as surrogate biomarkers to support clinical trials of targeted therapies and to pioneer a novel approach to CLAD diagnosis.

肺移植后长期生存的主要障碍是肺功能的逐渐丧失，称为慢性同种异体肺移植功能障碍（CLAD），其中小气道收缩性纤维化是一种病理性病症标志。 CLAD 在移植后 4 年内影响超过一半的肺移植受者，并且使大部分肺移植失败与移植相关的生活质量和功能改善。皮特、多伦多和加州大学旧金山分校肺科移植计划对肺移植的小气道刷检结果进行了完善的转录分析接受者作为一种新技术来了解 CLAD 所在解剖部位的基因表达变化病理发展。我们已经发表了与 CLAD 相关的基因表达变化，并经过我们的验证中心。该提案利用这种创新方法来了解 CLAD 发病机制。我们的初步数据显示气道刷洗中缺氧途径的早期上调，包括以下基因：在缺氧条件下使用气道上皮细胞培养物招募并激活细胞毒性 T 淋巴细胞气道刷转录组的通路分析。这种缺氧信号可能反映了紊乱微血管系统、支气管循环缺失以及与肺移植相关的血管炎症。在招募T淋巴细胞，我们还观察到肿瘤坏死因子超家族（TNFSF）基因的上调，它们是淋巴细胞靶标细胞凋亡的主要驱动因素。我们的数据显示气道俱乐部中优先发生细胞凋亡细胞，小气道的保护者和祖细胞，与上调的 TNF 相关细胞凋亡相关诱导配体（TRAIL）表达。我们在气道刷检和支气管肺泡方面的单细胞研究灌洗 (BAL) 液显示这些通路在上皮细胞和淋巴细胞类型之间的分离。基于根据这些数据，我们假设气道缺氧先于 TNFSF 基因表达和 T 细胞介导的气道俱乐部细胞凋亡驱动 CLAD 发病机制。为了检验这个假设，我们将生成平行队列研究 CLAD 的批量转录组和单细胞转录组与三个中心的对照相比，基因表达特征的严格交叉验证。我们将通过细胞培养来补充这些研究 - 基于方法来确定驱动这些基因表达变化的机制。在目标 1 中，我们将量化气道刷细胞中与 CLAD 相关的缺氧相关转录本，并确定缺氧如何促进淋巴细胞炎症。在目标 2 中，我们将确定 TNFSF 共刺激的细胞来源和动力学使用气道刷和 BAL 液在 CLAD 中进行分子表达。在目标 3 中，我们将调查 TRAIL 是否优先诱导俱乐部细胞凋亡。三大肺移植转化研究的协同作用具有世界一流的跨机构生物统计基础设施的计划提供了独特的机会严格地解决这个假设。 CLAD 随时间变化的细胞特异性基因表达特征通过这项研究产生的发展迫切需要作为替代生物标志物来支持靶向治疗的临床试验并开创了 CLAD 诊断的新方法。