Sputum-Based Profiling to Predict Cystic Fibrosis Exacerbations

基于痰液分析来预测囊性纤维化恶化

• 批准号: 10237397
• 负责人: Clemente Britto-Leon
• 金额: $ 8.38万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10237397
• 关键词: Acute; Acute Lung Injury; Adult; Airway Disease; Award; Bioinformatics; Biological Assay; Biological Markers; Cells; Chemistry; Clinical; Clinical Data; Clinical Trials; Cox Proportional Hazards Models; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Cystic Fibrosis sputum; Data; Data Analyses; Deterioration; Diagnosis; Diagnostic; Enzyme-Linked Immunosorbent Assay; Epithelial; Event; Exhibits; Funding; Future; Gene Expression; Genes; Genetic Diseases; Goals; Home; Hospitalization; Host Defense; Immune; Immunology; Individual; Influenza; Influenza A virus; Innate Immune Response; Interferons; Laboratories; Life; Link; Lung; Lung Inflammation; Lung diseases; Measures; Mentored Research Scientist Development Award; Methodology; Microbiology; Morbidity - disease rate; Mus; Mutation; Nasal Epithelium; National Heart, Lung, and Blood Institute; Palate; Patient Care; Patient Schedules; Patient Selection; Patients; Pharmaceutical Preparations; Property; Proteins; Pulmonary Function Test/Forced Expiratory Volume 1; Respiratory Tract Infections; Risk; Role; Sampling; Signal Transduction; Sputum; Subgroup; System; Testing; Therapeutic Intervention; Translating; United States; Visit; Work; airway inflammation; base; children with cystic fibrosis; clinically relevant; cohort; cystic fibrosis patients; high risk; immune activation; immunoregulation; macrophage; monocyte; mortality; neutrophil; pathogen; prevent; programs; pulmonary function; pulmonary function decline; receptor; respiratory; respiratory pathogen; single-cell RNA sequencing; therapeutic development; therapeutic target; tool; transcriptome; transcriptomics

PROJECT SUMMARY Cystic fibrosis (CF) is the most common life-shortening genetic disease in the United States. The primary cause of mortality in CF is lung disease, characterized by airway inflammation and lung function decline. CF acute exacerbations (AE) are episodes of rapid clinical deterioration, often associated with worsening airway inflammation and lung function. Limited data link sputum biomarkers and AE, but these markers are not predictive of future events. There are even fewer data analyzing sputum cellular components to diagnose or characterize AE. The lack of readily available noninvasive markers of AE is a critical unmet need, as they could guide therapeutic interventions to minimize morbidity, and facilitate patient selection for clinical trials in an era of accelerated therapeutic developments in CF. We previously showed that airway concentrations of host defense protein Short Palate Lung epithelium Clone 1 (SPLUNC1) are tightly regulated by airway inflammation and respiratory pathogens. Continuing this work on the soluble fraction of sputum, we show in preliminary data that SPLUNC1 is acutely decreased in AE and that stable CF subjects with low basal SPLUNC1 levels have a 6-fold increase in AE likelihood within 60 days. We are now integrating those studies with data from the cellular component of sputum, where preliminary data using single-cell RNA sequencing (scRNAseq) show that CF neutrophils and macrophages exhibit distinct transcriptomic profiles that separate AE from stable states. Our overall hypothesis is that a sputum panel combining soluble (SPLUNC1) and cellular (single- cell transcriptome profile) markers can robustly diagnose and predict early airway inflammation during AE in CF. We will test this using the following specific aims: Aim 1: Establish SPLUNC1 as a diagnostic tool and predictor of AE in CF; Aim 2: Define the AE-associated sputum single-cell transcriptome profile and its ability to predict AE; and Aim 3: Determine if a combined SPLUNC1-scRNAseq panel is a better predictor of AE than FEV1. This proposal is a translational application of mechanistic data generated during my K01 award. Our K01 work uncovered the immunomodulatory role of SPLUNC1 in lung host-pathogen interactions. The goal of this project is to investigate the link between SPLUNC1's immune properties and the sputum cell transcriptomic changes that occur during immune activation in AE. The R03 Award will support complementary studies for my NHLBI-K01-funded work as I apply for my first R01, focused on SPLUNC1-controlled mechanisms of acute lung injury during respiratory infections. The proposed work will translate our findings into clinically relevant tools that inform patient care and minimize the impact of AE for patients with CF and other airway diseases.

项目摘要 囊性纤维化（CF）是美国最常见的缩短寿命的遗传性疾病。主 CF的死亡原因是肺部疾病，其特征在于气道炎症和肺功能下降。cf 急性加重（AE）是临床快速恶化的发作，通常与气道恶化相关 炎症和肺功能。痰液生物标志物和AE之间的关联数据有限，但这些标志物不能预测 未来的事件。分析痰细胞成分来诊断或表征的数据就更少了 AE.缺乏现成的AE非侵入性标志物是一个关键的未满足的需求，因为它们可以指导 治疗干预措施，以尽量减少发病率，并促进患者的临床试验选择在一个时代， 加速了CF治疗的发展。 我们以前的研究表明，短腭肺上皮细胞中宿主防御蛋白的气道浓度 克隆1（SPLUNC1）受到气道炎症和呼吸道病原体的严格调控。继续这一 通过对痰液中可溶性成分的研究，我们在初步数据中显示，SPLUNC1在AE中急性减少， 并且具有低基础SPLUNC1水平的稳定CF受试者在60分钟内AE可能性增加6倍。 天我们现在正在将这些研究与痰液细胞成分的数据相结合， 使用单细胞RNA测序（scRNAseq）的数据显示CF中性粒细胞和巨噬细胞表现出不同的 将AE与稳定状态分开的转录组学特征。 我们的总体假设是，结合可溶性（SPLUNC1）和细胞（单细胞）的痰液组， 细胞转录组谱）标记物可以稳健地诊断和预测早期气道炎症， CF中的AE。我们将使用以下具体目标来测试这一点：目标1：将SPLUNC 1建立为诊断工具 和CF中AE的预测因素;目的2：定义AE相关的痰液单细胞转录组谱及其 目标3：确定组合SPLUNC1-scRNAseq组是否是AE的更好预测因子 FEV1。 这个建议是我在K01获奖期间产生的机械数据的翻译应用。我们的K01 工作揭示了SPLUNC1在肺宿主-病原体相互作用中的免疫调节作用。这个目标 一个项目是研究SPLUNC1的免疫特性和痰细胞转录组之间的联系， AE中免疫激活期间发生的变化。R03奖将支持我的补充研究， NHLBI-K01资助的工作，因为我申请了我的第一个R01，专注于SPLUNC1控制的急性肺损伤机制 呼吸道感染时受伤。拟议的工作将把我们的发现转化为临床相关的工具， 告知患者护理，并尽量减少CF和其他气道疾病患者的AE影响。

项目成果

SPLUNC1: a novel marker of cystic fibrosis exacerbations.

• DOI: 10.1183/13993003.00507-2020
• 发表时间: 2021-11
• 期刊: The European respiratory journal
• 作者: Khanal S;Webster M;Niu N;Zielonka J;Nunez M;Chupp G;Slade MD;Cohn L;Sauler M;Gomez JL;Tarran R;Sharma L;Dela Cruz CS;Egan M;Laguna T;Britto CJ
• 通讯作者: Britto CJ