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

• 批准号: 10597190
• 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
• 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/10597190
• 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; Gene Deletion; 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; 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; THBS1 gene; Technical Expertise; Testing; Thrombospondin 1; Tissue Banks; Tissue Sample; Training; Transgenic Mice; Transgenic Organisms; Universities; Uterus; Viral Respiratory Tract Infection; Virus Diseases; Wheezing; airway hyperresponsiveness; career; career development; cytokine; design; early childhood; experience; experimental study; extreme prematurity; 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; professor; profibrotic cytokine; recruit; response; skills; targeted treatment; transcriptome; transcriptome sequencing; transcriptomics; translational research program

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.

早产儿出生后会接触氧气 (O2)，这会导致长期发育不良 对肺部的影响。大约 70% 的极早产婴儿（胎龄 <29 周）会 肺部发病率和/或儿童早期喘息性疾病有所增加，尽管许多人并没有 诊断患有支气管肺发育不良（BPD）。这些婴儿特别容易受到呼吸道感染 呼吸道病毒感染后的高反应性（AHR）通过知之甚少的机制。在此，我们利用 基于新发现的低剂量高氧小鼠模型和独特的儿科人体组织储存库 新生儿的氧气会增加肺巨核细胞 (MK) 的丰度，这是一种尚未研究的骨髓细胞偏向性 朝向免疫调节功能。呼吸道病毒感染后，肺 MK 释放促纤维化细胞因子，例如 作为 Thrombospondin-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通过增加肺的 AHR 水平来为 AHR 做好准备。 感染后肺 MK 的募集和易感 MK 释放促纤维化因子（例如 TSP-1）。目的 1 将确定出生后高氧肺环境如何影响肺 MK 募集和播种，包括 不同发育年龄的 O2 和 MK 消耗如何影响肺 MK 群体。目标 2 将决定 使用体外细胞因子测定，新生儿高氧在激活前后如何影响肺 MK 转录组 和 RNA 测序。实验还将通过比较流感来确定 AHR 是否依赖于 MK 或 TSP-1 MK 缺失的小鼠感染 TSP-1 基因从 MK 中删除的转基因小鼠。目标3将 确定新生儿 O2 如何影响骨髓 MK 池，包括其对血小板生成的影响。 该提案是医学博士 Andrew Dylag 博士的一项为期五年的指导研究奖和培训计划 研究小鼠和获得的人体组织中氧诱导的气道功能障碍的机制。博士。 Dylag 是罗彻斯特大学医学中心儿科（新生儿学）助理教授。这 本文的研究建立在 Dylag 博士作为临床新生儿学家和基础科学家的经验之上 O2 损伤和高氧后气道高反应性 (AHR)。作为其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) 提高调查方面的知识和技术技能 使用转化体内模型进行肺发育和损伤后修复，2）生物信息学定向训练 分析，包括转录组学，3) 发展将体内实验室发现应用于人类的专业知识 组织和临床人类疾病，以及 4) 培养领导有效转化研究的必要技能 程序。 Dylag 博士将通过应用流式细胞术、免疫组织化学、 转录组学，以及针对小鼠和人体组织的有针对性的生物信息学培训。完成本次操作后 职业发展奖，Dylag 博士将探讨生命早期 O2 如何驱动 AHR 的一种机械作用， 增进我们对新生儿氧气暴露如何导致长期肺部发病率的理解。