Sputum-Based Profiling to Predict Cystic Fibrosis Exacerbations

项目总结 囊性纤维化(CF)是美国最常见的缩短寿命的遗传病。初级阶段 CF的死亡原因是肺部疾病，其特征是呼吸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，通常与呼吸道恶化有关 炎症和肺功能。有限的数据链接了痰生物标记物和AE，但这些标记物不能预测 关于未来的事件。分析痰细胞成分来诊断或表征的数据就更少了 天哪。缺乏现成的非侵入性AE标志物是一个严重的未得到满足的需求，因为它们可以引导 治疗干预措施，以最大限度地减少发病率，并促进在一个 加速了CF的治疗进展。 我们先前的研究表明，呼吸道内宿主防御蛋白的浓度与短腭肺上皮 克隆1(SPLunc1)受呼吸道炎症和呼吸道病原体的严格调控。继续这样做 在对痰中可溶部分的研究中，我们在初步数据中显示，SPLunc1在AE中显著降低 而基础水平较低的稳定的慢性充血性心力衰竭患者在60岁内发生脑梗塞的可能性增加了6倍 几天。我们现在正在将这些研究与来自痰细胞成分的数据结合起来，在那里初步 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 将声发射与稳定状态分开的转录图谱。 我们的总体假设是，一个结合了可溶性(SPLunc1)和细胞(单一-1)的痰标本 细胞转录组)标志物可以强有力地诊断和预测早期呼吸道炎症 Cf中的声发射我们将使用以下具体目标对其进行测试：目标1：将SPLunc1建立为诊断工具 和预测因子；目标2：确定与AE相关的痰单细胞转录组图谱及其 预测AE的能力；和目标3：确定SPLunc1-scRNAseq组合是否是预测AE的更好指标 而不是FEV1。 这份提案是对我在K01颁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在肺部宿主-病原体相互作用中的免疫调节作用。这样做的目的是 项目是研究SPLUN1的S免疫特性与痰细胞转录之间的联系 在AE的免疫激活过程中发生的变化。R03奖将支持我的补充学习 NHLBI-K01资助的工作，因为我申请了我的第一个R01，专注于SPLunc1控制的急性肺机制 在呼吸道感染期间受伤。这项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，最大限度地减少AE对CF和其他呼吸道疾病患者的影响。

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.