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：
10521842
负责人：
JOHN GREENLAND
金额：
$ 64.52万
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/10521842
关键词：
Address Affect Airway Fibrosis Alloantigen Anatomy Apoptosis Apoptotic Automobile Driving Bioinformatics Biometry Blood Circulation 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 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 Gene Proteins Genes Genetic Transcription Heat shock proteins Hypoxia Hypoxia Pathway Image Immune Immune response Immunity Immunobiology In Vitro Inflammation Infrastructure Investigation Kinetics Lead Ligands Link Lung Lung Transplantation Lung diseases Lymphocyte Lymphoid Cell Mediating Methods OX40 Pathogenesis Pathologic Pathology Pathway Analysis Pathway interactions Phenotype Play Prevention Primary Cell Cultures Publishing Quality of life Reporting Role Signal Transduction Source Stimulus Stress Structure of parenchyma of lung Surrogate Markers T cell response T-Lymphocyte 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 base cell type chemokine clinical phenotype clinical subtypes cohort constriction cytokine cytotoxic effector T cell fibrotic lung functional improvement improved innovation insight lung allograft macrophage 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.

肺移植后长期存活的主要障碍是肺功能的进行性丧失，称为慢性肺同种异体功能障碍（CLAD），对于小气道中的约束纤维化是一种病理标志。移植后4年影响肺移植受者的一半以上，并否定了大部分与移植相关的生活质量和功能改善。皮特，多伦多和UCSF肺移植程序已经完善了对肺移植的小气道刷子的转录分析接受者是一种新的技术，可以理解基因表达在解剖部位变化病理发展。我们已经发表了与我们在我们的整个验证的基因表达变化相关的基因表达变化中心。该提议利用这种创新的方法来了解层层发病机理的机制。我们的初步数据表明，在气道刷刷中缺氧途径的早期上调，包括在低氧条件和气道刷转录组的途径分析。这种缺氧信号传导可能反映了无序微脉管系统，没有支气管循环以及与肺移植有关的血管炎症。在招募的T淋巴细胞，我们还观察到肿瘤坏死因子超家族（TNFSF）基因上调，这是淋巴细胞靶标的凋亡的主要驱动因素。我们的数据显示气道俱乐部的优先凋亡小型气道的细胞，保护者和祖细胞，与TNF相关的凋亡上调 - 诱导配体（TRAIL）表达。我们在气道刷子和支气管肺泡中进行的单细胞调查灌洗液（BAL）流体显示这些途径在上皮和淋巴样细胞类型上的分离。基于这些数据，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着甲壳虫的发病机理。为了检验这一假设，我们将生成平行队列研究三个中心的外壳与对照组的批量和单细胞转录组，从而允许严格的基因表达特征的交叉验证。我们将通过细胞培养来补充这些研究 - 基于确定驱动这些基因表达的机制变化的方法。在AIM 1中，我们将量化相对于外壳，气道刷细胞中与缺氧相关的转录本，并确定缺氧如何促进淋巴细胞炎症。在AIM 2中，我们将确定TNFSF共刺激性的细胞来源和动力学使用气道刷和BAL流体在外壳中的分子表达。在AIM 3中，我们将调查是否跟踪优先诱导俱乐部细胞凋亡。三个大型肺移植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为您提供了一个独特的机会严格解决这一假设。在覆盖时间过程中的细胞特异性基因表达签名作为替代生物标志物来支持，这项研究将产生的开发非常需要靶向疗法的临床试验和开创一种新型诊断方法的新方法。