The Role of Lung Megakaryocytes in Airway Disease after Neonatal Hyperoxia

肺巨核细胞在新生儿高氧后气道疾病中的作用

基本信息

批准号：
10448727
负责人：
Andrew Michael Dylag
金额：
$ 16.21万
依托单位：
UNIVERSITY OF ROCHESTER
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：
2022-04-01 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10448727
关键词：
Address Adult Affect Age Airway Disease Antibodies Autopsy Award Bioinformatics Biological Assay Birth Blood Platelets Bone Marrow Breathing Bronchopulmonary Dysplasia Caring Cell Lineage Cells Childhood Clinical Development Development Plans Disease Dose Effector Cell Environment Exposure to Fibrosis Flow Cytometry Functional disorder Future Genes Gestational Age Glycoproteins Goals Healthcare Systems Human Hyperoxia Immune Immunohistochemistry Immunophenotyping In Vitro Incidence Infant Infection Inflammatory Influenza A Virus, H3N2 Subtype Influenza A virus Injury Investigation K-Series Research Career Programs Knowledge Label Laboratories Lead Life Lung Lung diseases Mechanical ventilation Mediator of activation protein Medical center Megakaryocytes Mentors Modeling Morbidity - disease rate Morphology Mus Myelogenous Myeloid Cells Myofibroblast Neonatal Neonatal Hyperoxic Injury Neonatology Oxygen Oxygen Therapy Care Pediatrics Phenotype Population Pregnancy Premature Birth Premature Infant Production Profibrotic signal Research Respiratory Tract Infections Role Scientist Severities TGFB1 gene Technical Expertise Testing Thrombospondin 1 Tissue Banks Tissue Sample Training Transgenic Mice Transgenic Organisms Universities Viral Respiratory Tract Infection Virus Diseases Wheezing airway hyperresponsiveness career career development cytokine design early childhood experience experimental study hospital readmission human disease human tissue immunoregulation improved in vivo in vivo Model influenza infection interest lung development lung repair mouse model novel pathogen platelet function premature professor profibrotic cytokine recruit response skills skills training targeted treatment transcriptome transcriptome sequencing transcriptomics translational research program

项目摘要

Former preterm infants are exposed to oxygen (O2) after birth which results in long-term developmental impacts on the lung. Approximately 70% of infants born extremely prematurely (<29 weeks’ gestational age) will have increased pulmonary morbidity and/or early childhood wheezing disorders even though many are not diagnosed with Bronchopulmonary Dysplasia (BPD). These infants are especially vulnerable to airway hyperreactivity (AHR) after respiratory viral infections through poorly understood mechanisms. Herein, we utilize a low-dose hyperoxia mouse model and a unique pediatric human tissue repository grounded on a new discovery that neonatal O2 increases the abundance of lung megakaryocytes (MKs), an understudied myeloid cell biased toward immunomodulatory functions. After respiratory viral infection, lung MKs release profibrotic cytokines such as Thrombospondin-1 (TSP-1), a critical inflammatory regulator and activator of transforming growth factor beta 1 (TGFβ1) that drives fibrosis. We hypothesize that neonatal hyperoxia primes the lung for AHR by increasing the recruitment of lung MKs and predisposing MKs to release pro-fibrotic factors (e.g. TSP-1) after infection. Aim 1 will determine how the hyperoxic lung environment after birth effects lung MK recruitment and seeding including how O2 at different developmental ages and MK depletion affect the lung MK population. Aim 2 will determine how neonatal hyperoxia effects lung MK transcriptome before and after activation using in vitro cytokine assays and RNAseq. Experiments will also determine if AHR is MK or TSP-1 dependent by comparing Influenza infection models of MK-depleted mice to transgenic mice with the TSP-1 gene deleted from MKs. Aim 3 will determine how neonatal O2 effects the bone marrow MK pool, including its effects on platelet production. This proposal is a five-year mentored research award and training plan for Dr. Andrew Dylag, MD to investigate oxygen-induced mechanisms of airway dysfunction in both mice and procured human tissues. Dr. Dylag is an Assistant Professor of Pediatrics (Neonatology) at the University of Rochester Medical Center. The research herein builds on Dr. Dylag’s experience as a clinical neonatologist and a basic scientist interested in O2 injury and post-hyperoxia airway hyperreactivity (AHR). As part of his career development plan, Dr. Dylag will attain expertise through four (4) career aims: 1) Increase knowledge and technical skills in the investigation of lung development and repair after injury using translational in vivo models, 2) Targeted training in bioinformatics analysis including transcriptomics, 3) Develop expertise in applying in vivo laboratory discoveries to human tissues and clinical human disease, and 4) Develop the necessary skills to lead an effective translational research program. Dr. Dylag will attain his stated goals by applying new skills in flow cytometry, immunohistochemistry, transcriptomics, and targeted bioinformatics training to a mouse and human tissues. At the completion of this career development award, Dr. Dylag will have interrogated one mechanistic role of how early life O2 drives AHR, advancing our understanding of how neonatal O2 exposures drive longer-term pulmonary morbidity.

出生后，以前的早产儿就暴露于氧气（O2），导致长期发育对肺的影响。大约70％的婴儿过早出生（胎龄<29周）将肺部发病率和/或幼儿时期喘不过气，即使许多人不是被诊断为支气管肺发育不良（BPD）。这些婴儿特别容易受到气道的影响通过较知的机制，呼吸道病毒感染后的过度反应性（AHR）。在此，我们利用低剂量的高氧小鼠模型和一个独特的儿科人体组织存储库，该存储库接地于新发现新生儿O2增加了肺巨核细胞（MK）的抽象，一种理解的髓样细胞有偏见迈向免疫调节功能。呼吸道病毒感染后，肺MK释放纤维化细胞因子这样作为血小板传播-1（TSP-1），一种关键的炎症调节剂和转化生长因子β的激活剂 1（TGFβ1）驱动纤维化。我们假设新生儿高氧素质通过增加而增加了AHR的肺感染后，募集肺MK和诱发MK释放促纤维化因子（例如TSP-1）。目的 1将确定出生后的高氧肺环境如何肺MK募集和种子，包括 O2在不同的发育年龄和MK部署如何影响肺MK人群。 AIM 2将确定新生儿高氧如何使用体外细胞因子测定前后对肺MK转录组有何影响和rnaseq。实验还将通过比较流感来确定AHR是MK还是TSP-1依赖用MK从MKS中删除的TSP-1基因的MK耗尽小鼠的感染模型。目标3意志确定新生儿O2如何影响骨髓MK池，包括其对血小板产生的影响。该提案是为医学博士Andrew Dylag博士致敬的五年指导研究奖和培训计划研究小鼠和采购的人体组织中气道功能障碍的氧诱导的机制。博士 Dylag是罗切斯特大学医学中心的儿科助理教授（新生儿学）。这这里的研究基于Dylag博士作为临床新生儿学家的经验和对此感兴趣的基础科学家 O2损伤和休毒后气道高反应性（AHR）。作为他职业发展计划的一部分，Dylag博士将通过四（4）个职业目标获得专业知识：1）在调查中提高知识和技术技能使用体内翻译模型受伤后的肺发育和修复，2）对生物信息学的靶向培训分析包括转录组学，3）在将体内实验室发现应用于人类方面发展专业知识组织和临床人类疾病，以及4）发展必要的技能以领导有效的转化研究程序。 Dylag博士将通过运用流式细胞仪，免疫组织化学的新技能来实现他的既定目标转录组学，并针对小鼠和人体组织进行生物信息学训练。完成此过程职业发展奖，Dylag博士将询问O2早期生活如何驱动AHR的一种机械作用，促进我们对新生儿O2暴露如何驱动长期肺部发病率的理解。