Novel exosomal niches for alveolar stem cell-bassed repair of ARDS

用于 ARDS 肺泡干细胞修复的新型外泌体生态位

基本信息

批准号：
10836707
负责人：
HONG-LONG JI
金额：
$ 48.02万
依托单位：
LOYOLA UNIVERSITY CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10836707
关键词：
3-Dimensional Acute Respiratory Distress Syndrome Alveolar Bacterial Infections Bass Bioinformatics Biological Markers Biological Specimen Banks Biomedical Engineering Biometry Bronchoalveolar Lavage Bronchoalveolar Lavage Fluid Catalysis Cell Lineage Cells Classification Clinical Clinical Data Clinical Research Clinical Trials Data Disease Double-Blind Method Drug Targeting Early Diagnosis Enzymes Epithelial Cells Epithelium Etiology Gastrointestinal tract structure Goals Hospital Mortality Human Impairment In Vitro Infection Influenza Irrigation Kidney Liver Lung Mediating Mediator Methodology Modeling Multiple Organ Failure Mus National Heart, Lung, and Blood Institute Natural regeneration Organ Organoids Outcome Oxygen Paracrine Communication Pathogenesis Pathologic Processes Pathway interactions Patients Pharmacologic Substance Pharmacy (field)Pneumonia Process Prognosis Proteins Proteomics Randomized Controlled Clinical Trials Recovery Regulation Role Sampling Sepsis Septic Shock Severities Signal Pathway Signal Transduction Signaling Molecule Stratification Testing Tissues United States National Institutes of Health Validation Virus Diseases alveolar epithelium autocrine biomarker identification candidate marker cohort data repository design differential expression druggable target epithelial injury epithelial stem cell epithelium regeneration exosome extracellular extracellular vesicles in vivo in vivo Model injury and repair lung injury lung regeneration lung repair machine learning algorithm mortality novel novel marker organ injury paracrine preclinical study protein biomarkers regenerative repaired sepsis induced ARDS septic stem cell fate stem cell therapy stem cells success supervised learning urogenital tract

项目摘要

Summary. The regulation of alveolar regeneration of injured lungs by exosomal signals in acute respiratory distress syndrome (ARDS) are incompletely studied. We have identified exosomal proteins in bronchoalveolar lavage fluid (BALF) of ARDS patients that are closely association with the severity of lung injury. In ARDS, clinical severity is graded by the ratio of arterial oxygen tension (PaO2 in mmHg) to the fraction of inspired oxygen (FiO2). We have identified stem cell-related niche proteins that are present in mild (200£PaO2/FiO2<300 mmHg) to moderate ARDS (100£PaO2/FiO2<200 mmHg) but reduced in severe ARDS (PaO2/FiO2<100 mmHg). A key pathological process in ARDS is damage to the alveolar epithelium, which has been demonstrated in numerous preclinical and clinical studies to be associated with ARDS severity. Therefore, we hypothesize that exosomal signals for lung stem cell-mediated re-alveolarization are determinants of ARDS outcomes. Several findings support this hypothesis. 1) Exosomes are effective autocrine/paracrine pathways for stem cell lineage in injured organs. 2) Exosomal proteins are protected from catalysis by enzymes in inflamed tissues. 3) Unbiased high throughput proteomic analysis and advanced bioinformatic platforms have successfully identified novel biomarkers for other diseases. 4) Although the etiologies of ARDS are diverse, the repair processes mediated by epithelial stem/progenitor cells regulated by niches seem to be similar based on preclinical studies. Our objective is to test this hypothesis with BALF samples and patients' clinical data from NIH/NHLBI-supported clinical trials, prioritize niche molecules, and validate the results in genetically bioengineered mice and organoids of alveolar type 2 (AT2) epithelial cells. We will apply novel “exosomics” approaches, cutting-edge bioinformatics, three-dimensional culture models, robust cell origin tracking, and supervised machine learning algorithms. There are three specific aims: Aim 1 is designed to identify exosomal signaling pathways and networks in lavage that regulate the lineage of lung stem cells for re-alveolarization. We hypothesize that stem cell-mediated re- alveolarization has been suppressed in severe ARDS patients due to the disruption of key exosomal niches. We will prioritize differentially expressed exosomal proteins and related signaling pathways and networks using R packages. Aim 2 is designed to detect and optimize regenerative predictors for re-alveolarization of injured lungs. We hypothesize that the significantly differential exosomal molecules in lavage will be associated with clinical data in ARDS patients. We will perform unbiased clustering and supervised machine learning algorithms to develop models for discovering critical signals for lung regeneration. The prioritized exosomal predictors will be compared with traditional whole-protein markers for accuracy and applicability. Aim 3 will validate selected exosomal signals for AT2-mediated re-alveolarization. We will confirm that the exosomal molecules regulate the lineage of human and mouse AT2 cells in 3D organoids and mice. These studies will identify stem cell-specific exosomal molecules as potential pharmaceutic targets for ARDS.

摘要：急性呼吸道疾病中外泌体信号对受损肺肺泡再生的调节我们已经鉴定出支气管肺泡中的外泌体蛋白。 ARDS患者的灌洗液（BALF）与ARDS的临床肺损伤严重程度密切相关。严重程度根据动脉氧分压（PaO2，mmHg）与吸入氧分数（FiO2）的比率进行分级。我们已经鉴定出存在于轻度（200£PaO2/FiO2<300 mmHg）至中度ARDS（100£PaO2/FiO2<200 mmHg），但重度ARDS（PaO2/FiO2<100 mmHg）则减少。 ARDS 的病理过程是肺泡上皮的损伤，这已在许多研究中得到证实。因此，我们面临着外泌体与 ARDS 严重程度相关的临床前和临床研究。肺干细胞介导的再肺泡化信号是 ARDS 结局的决定因素。研究结果支持这一假设。1) 外泌体是干细胞谱系的有效自分泌/旁分泌途径。 2) 外泌体蛋白免受炎症组织中酶的催化。高通量蛋白质组分析和先进的生物信息平台已成功鉴定出新的 4) 尽管 ARDS 的病因多种多样，但修复过程有所介导。根据我们的临床前研究，上皮干/祖细胞受生态位调节的作用似乎相似。目的是用 BALF 样本和来自 NIH/NHLBI 支持的患者临床数据来检验这一假设临床试验，优先考虑利基分子，并验证基因生物工程小鼠和类器官的结果我们将应用新型“外泌体”方法、尖端生物信息学、三维培养模型、强大的细胞起源跟踪和监督机器学习算法。是三个具体目标：目标 1 旨在识别灌洗液中的外泌体信号通路和网络，调节肺干细胞谱系以重新肺泡化。由于关键外泌体生态位的破坏，严重 ARDS 患者的肺泡化受到抑制。将使用 R 优先考虑差异表达的外泌体蛋白以及相关信号通路和网络 Aim 2 旨在检测和优化受损肺部重新肺泡化的再生预测因子。我们追求灌洗液中显着差异的外泌体分子与临床相关我们将执行无偏聚类和监督机器学习算法来开发模型来发现肺再生的关键信号将是优先考虑的外泌体预测因子。与传统的全蛋白标记物相比，Aim 3 将验证所选的准确性和适用性。 AT2 介导的再肺泡化的外泌体信号我们将证实外泌体分子调节 3D 类器官和小鼠中人类和小鼠 AT2 细胞的谱系这些研究将鉴定干细胞特异性。外泌体分子作为 ARDS 的潜在药物靶点。