S-nitrosoglutathione reductase and airways hyperreactivity in murine bronchopulmonary dysplasia

S-亚硝基谷胱甘肽还原酶和小鼠支气管肺发育不良中的气道高反应性

基本信息

批准号：
9975210
负责人：
Thomas Michael Raffay
金额：
$ 16.42万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
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：
2017-07-14 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9975210
关键词：
Acute Adolescent Adult Aerosols Agonist Air Airway Disease Alveolar Animal Model Anti-Inflammatory Agents Asthma Attenuated Biology Breathing Bronchial Spasm Bronchoconstriction Bronchodilator Agents Bronchopulmonary Dysplasia Catabolism Cell Separation Cells Child Childhood Chronic Clinic Clinical Clinical Trials Complication Cystic Fibrosis Data Development Diagnosis Dose Down-Regulation Educational process of instructing Enzymes Exposure to Extramural Activities Extrinsic asthma Flow Cytometry Fluorescence-Activated Cell Sorting Gases Human Hyperoxia Incidence Infant Inhalation International Knock-out Knockout Mice Lead Life Location LoxP-flanked allele Lung Lung diseases Measures Mentors Mentorship MicroRNAs Modeling Mus Neonatal Neonatal Hyperoxic Injury Newborn Infant Oxidoreductase Oxygen Pediatric Hospitals Pharmaceutical Preparations Pharmacologic Substance Pharmacology Phenotype Premature Birth Premature Infant Property Research Research Personnel Research Training S-Nitrosoglutathione S-Nitrosothiols Scientist Severities Signal Transduction Site Smooth Muscle Structure Structure of parenchyma of lung Supplementation Survivors Testing Time Tissues Training Transgenic Animals Transgenic Mice Translational Research United States Universities Wheezing Wild Type Mouse aerosolized airway hyperresponsiveness airway obstruction attenuation base career development conditional knockout cost enzyme activity experience fetal improved inhibitor/antagonist laboratory experience mouse model novel overexpression prevent protein expression pup respiratory respiratory smooth muscle scaffold skills training socioeconomics

项目摘要

Project Abstract/Summary: This proposal describes a five-year mentored research and training plan that will facilitate the development of Dr. Thomas Raffay, MD to an independent investigator in neonatal pulmonary disease. Building upon Dr. Raffay's background as a clinical neonatologist and a basic scientist in airways reactivity and neonatal bronchopulmonary dysplasia (BPD), he will attain expertise in S-nitrosothiol biology, flow-cytometry, conditional transgenic animal models, pharmacology, and translational science through structured mentorship, rigorous hands-on laboratory experiences, didactics teaching, and formal classwork and skills training at Case Western Reserve University/Rainbow Babies and Children's Hospital, the Mayo Clinic Rochester, and the University of Windsor. Dr. Benjamin Gaston, a pioneer in S-nitrosothiol signaling in pediatric lung disease, and Dr. Richard J Martin, an international leader in neonatal airways disease and BPD, will provide their expertise and an established track record of mentorship to this project and Dr. Raffay's transition to research independence. In the United States, 14,000 new diagnoses of the pediatric lung disease, BPD, are made annually in surviving premature infants, with costs exceeding $2.5 billion. Treatment options are limited for the severe bronchospasms and life-long airway obstruction that characterize BPD. Using a neonatal hyperoxia mouse model of BPD and airways hyperreactivity, Dr. Raffay's new data identify a viable therapy. Treatment with a single aerosol of S-nitrosoglutathione (GSNO) reverses airways hyperresponsiveness in juvenile BPD mice and room air recovered adults. GSNO is a potent endogenous bronchodilator, critical for the airways diseases of asthma and cystic fibrosis. In this model, neonatal hyperoxia increases the catabolic breakdown of GSNO caused by increased expression and activity of the enzyme, S-nitrosoglutathione reductase (GSNOR), at least in part through hyperoxic downregulation of a microRNA (miR-342-3p). This study will test the overall hypothesis that loss or inhibition of GSNOR activity will protect against BPD airways hyperreactivity in juvenile and adult mice. This proposal will evaluate the effects of neonatal hyperoxia on BPD airways hyperreactivity and parenchymal lung remodeling utilizing global GSNOR knockout mice (Aim 1A); identify the lung tissues involved in GSNOR and miR-342-3p expression utilizing fluorescence-activated cell sorting and develop and test conditional GSNOR knockout mice (Aim 1B); test a pharmaceutical-grade GSNOR inhibitor drug in wild- type mice to alleviate acute bronchoconstriction (Aim 2a) and chronically to attenuate the BPD phenotype (Aim 2b). In a translational human aim (Aim 3): GSNOR levels, real-time fluorescent GSNO catabolism, and microRNA expression will be measured in human fetal airway smooth muscle exposed to hyperoxia; and GSNO and GSNOR inhibitors will be tested as a treatment for attenuation of oxygen induced hyper- contractility. These studies are clinically important because GSNOR inhibitors are currently in clinical trials for cystic fibrosis and asthma which could ultimately serve as a new treatment for bronchopulmonary dysplasia.

项目摘要/摘要：该提案描述了一个为期五年的指导研究和培训计划，该计划将促进发展 Thomas Raffay 博士是一位新生儿肺病独立研究者的医学博士。以博士为基础 Raffay 作为临床新生儿学家和气道反应性和新生儿基础科学家的背景支气管肺发育不良 (BPD)，他将获得 S-亚硝基硫醇生物学、流式细胞术、条件性通过结构化的指导、严格的指导，转基因动物模型、药理学和转化科学凯斯西储大学的实验室实践经验、教学法以及正式的课堂作业和技能培训储备大学/彩虹婴儿和儿童医院、罗切斯特梅奥诊所和罗切斯特大学温莎。 Benjamin Gaston 博士（小儿肺部疾病 S-亚硝基硫醇信号传导领域的先驱）和 Richard J 博士 Martin 是新生儿气道疾病和 BPD 领域的国际领导者，他将提供他们的专业知识和建立了对该项目的指导以及 Raffay 博士向研究独立过渡的跟踪记录。在在美国，每年新诊断出 14,000 例儿童肺病 (BPD) 早产儿的费用超过 25 亿美元。重症患者的治疗选择有限支气管痉挛和终生气道阻塞是 BPD 的特征。使用新生高氧小鼠通过 BPD 和气道高反应性模型，Raffay 博士的新数据确定了一种可行的治疗方法。治疗用 S-亚硝基谷胱甘肽 (GSNO) 单一气雾剂可逆转幼年 BPD 小鼠的气道高反应性和室内空气恢复成人。 GSNO 是一种有效的内源性支气管扩张剂，对气道疾病至关重要哮喘和囊性纤维化。在该模型中，新生儿高氧会增加 GSNO 的分解代谢分解至少由 S-亚硝基谷胱甘肽还原酶 (GSNOR) 的表达和活性增加引起部分是通过高氧下调 microRNA (miR-342-3p)。本研究将测试总体假设 GSNOR 活性的丧失或抑制将防止青少年 BPD 气道高反应性和成年小鼠。该提案将评估新生儿高氧对 BPD 气道高反应性的影响利用全局 GSNOR 基因敲除小鼠进行肺实质重塑（目标 1A）；识别肺组织利用荧光激活细胞分选参与 GSNOR 和 miR-342-3p 表达并开发和测试条件 GSNOR 基因敲除小鼠（目标 1B）；在野外测试药物级 GSNOR 抑制剂药物型小鼠缓解急性支气管收缩（目标 2a）并长期减弱 BPD 表型（目标 2b）。人类转化目标（目标 3）：GSNOR 水平、实时荧光 GSNO 分解代谢和将在暴露于高氧的人胎儿气道平滑肌中测量 microRNA 表达；和 GSNO 和 GSNOR 抑制剂将作为减弱氧诱导的高氧血症的治疗方法进行测试。收缩性。这些研究具有重要的临床意义，因为 GSNOR 抑制剂目前正在进行临床试验囊性纤维化和哮喘，最终可能成为支气管肺发育不良的新治疗方法。