Mechanisms of Pulmonary Microbiota-Induced Inflammation and Vascular Dysfunction in Neonatal Lung Injury

新生儿肺损伤中肺部微生物群引起的炎症和血管功能障碍的机制

• 批准号: 10355431
• 负责人: Charitharth Vivek Lal
• 金额: $ 16.76万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-03 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10355431
• 关键词: Advisory Committees; Aspirate substance; Attenuated; Award; Bacteria; Bioinformatics; Biology; Birth; Blood Vessels; Bronchopulmonary Dysplasia; Chronic; Chronic lung disease; Collagen; Data; Development; Development Plans; Disease; Endothelium; Enzymes; Epithelial Cells; Extracellular Matrix; Extremely Low Birth Weight Infant; Foundations; Funding; Gammaproteobacteria; Gelatinase B; Germ-Free; Goals; Human; In Vitro; Infant; Inflammation; Inflammatory; Injury; Knowledge; Lead; Leukocytes; Lipopolysaccharides; Lung; Mediating; Mentors; MicroRNAs; Molecular Biology; Morbidity - disease rate; Mus; N-acetyl-proline-glycine-proline; Neonatal; Newborn Infant; Pathogenesis; Pattern; Peptide Hydrolases; Permeability; Phenotype; Physicians; Play; Positioning Attribute; Pregnancy; Premature Infant; Production; Prospective cohort study; Protein Fragment; Proteobacteria; Publishing; Pulmonary Inflammation; Research; Research Ethics; Research Personnel; Research Proposals; Role; Sampling; Scientist; Testing; Training; United States National Institutes of Health; Untranslated RNA; Validation; Vascular Diseases; Work; career; career development; cohort; didactic education; dysbiosis; early detection biomarkers; exosome; gain of function; human study; in vitro Model; in vivo; lung microbiome; lung microbiota; microbial; microbiome; microbiome research; microbiome sequencing; microbiota; mouse model; neonatal lung injury; neutrophil; novel; predictive modeling; programs; prolyl oligopeptidase; respiratory microbiome; skills; therapeutic miRNA

PROJECT SUMMARY/ABSTRACT: This revised NIH K08 proposal describes a 5-year training and research plan for the candidate, a physician scientist with a long term goal of becoming an independent investigator in the field of neonatal pulmonary biology with expertise in understanding the role of the pulmonary microbiome in chronic lung disease of prematurity. To accomplish this goal, he and his mentoring committee put forth an integrated career development plan encompassing a novel research idea. Bronchopulmonary dysplasia (BPD), the most common pulmonary morbidity in extremely preterm infants is initiated by injury to the immature lung by early neutrophil influx, collagen degradation, remodeling, and arterial thickening. The candidate has discovered that the airways of newborn infants are not sterile but are occupied by a diverse microbiome even at birth, and that the microbiome is altered (dysbiosis) during the development of BPD. He has also recently discovered the role of exosomal microRNAs in BPD prediction and pathogenesis. His mentors have previously established that the tripeptide N-acetyl proline-glycine-proline (Ac-PGP) derived from the breakdown of the ECM plays a critical role in various chronic lung diseases by enhancing neutrophilic inflammation and endothelial permeability. The preliminary data in this proposal indicate that the airways of infants with BPD which have increased Gammaproteobacteria (¡-Proteobacteria) also have increased levels of Ac-PGP. Moreover, gain of Ac-PGP function creates the phenotype of BPD in murine models, whereas loss of Ac-PGP function reverses the BPD phenotype. Matrix metalloproteinase 9 (MMP9) and prolyl endopeptidase (PE) degrade collagen, leading to the release of Ac-PGP. MMP9 and PE are top predicted targets of exosomal miR 548m and miR 129-1-3 respectively and both these miRs are reduced in BPD. Collectively, these findings lead to the novel mechanistic hypothesis that ¡-Proteobacteria-induced reduction in exosomal miRs increase protease levels which in turn increase Ac-PGP release and cause chronic neutrophilic inflammation and vascular dysfunction in BPD. In addition to determining these mechanisms (Aim 1), the candidate will conduct a human study using independent ‘Discovery’ and ‘Validation’ cohorts of extremely preterm infants to test the hypothesis that decreased exosomal miRs 548m and miR 129- 1-3p and increased MMP9, PE and Ac-PGP, in relation to a dysbiotic airway microbota are early predictors of severe BPD (Aim 2). Thus the work proposed in this research proposal will generate novel information about the mechanisms of microbiota induced neutrophilic inflammation and vascular dysfunction in BPD, and will determine novel early biomarkers for BPD. The candidate has already assembled a research advisory committee with complementary intellect and content expertise that can guide him throughout the award period. In addition, the candidate will obtain didactic education to gain requisite knowledge culminating in the awarding of an MSPH degree. Through this integrated mentoring and didactic plan, the candidate will gain skill and expertise in pulmonary inflammation, microbial pathogenesis, microRNA and exosomal molecular biology, microbiome sequencing, bioinformatics, skills in manipulation of germ free mouse models and research ethics. This will enable the him to develop an independent research program and obtain NIH R01 funding.

项目总结/摘要： 这个修订的NIH K 08提案描述了一个为期5年的培训和研究计划的候选人，一名医生 科学家，长期目标是成为新生儿肺部疾病领域的独立研究者 生物学，在了解肺部微生物组在慢性肺部疾病中的作用方面具有专业知识， 早产为了实现这一目标，他和他的指导委员会提出了一个综合的职业生涯 发展计划包含一个新的研究思路。 支气管肺发育不良（BPD）是极早产儿最常见的肺部疾病， 由早期中性粒细胞流入、胶原降解、重塑和动脉粥样硬化引起的未成熟肺损伤引发 增厚候选人已经发现，新生儿的气道不是无菌的，而是被 即使在出生时也有多样化的微生物组，并且微生物组在发育过程中发生改变（生态失调）。 波士顿警局他最近还发现了外泌体microRNA在BPD预测和发病机制中的作用。 他的导师先前已经确定，三肽N-乙酰脯氨酸-甘氨酸-脯氨酸（Ac-PGP）衍生 从ECM的分解起着关键作用，在各种慢性肺部疾病，通过提高嗜酸性粒细胞 炎症和内皮渗透性。该提案中的初步数据表明， γ-变形菌门（γ-Proteobacteria）增加的BPD婴儿， Ac-PGP。此外，Ac-PGP功能的获得在小鼠模型中产生BPD表型，而Ac-PGP功能的丧失在小鼠模型中产生BPD表型。 Ac-PGP功能逆转BPD表型。基质金属蛋白酶9和脯氨酰内肽酶 (PE)降解胶原蛋白，导致Ac-PGP的释放。MMP 9和PE是外泌体的最佳预测靶点。 miR 548 m和miR 129-1-3分别是相同的，并且这两种miR在BPD中均降低。总的来说，这些发现 导致了新的机制假说，即β-变形菌诱导外泌体miR减少 蛋白酶水平增加，从而增加Ac-PGP释放并导致慢性中性粒细胞增多 炎症和血管功能障碍。除了确定这些机制（目标1）外， 候选人将使用独立的“发现”和“验证”队列进行人体研究， 早产儿来检验外泌体miR 548 m和miR 129- 1- 3 p减少以及miR 129- 1- 3 p增加的假设。 MMP 9、PE和Ac-PGP与气道微生物菌群失调有关，是严重BPD的早期预测因子（目的2）。 因此，这项研究提案中提出的工作将产生关于以下机制的新信息： 微生物群诱导BPD的嗜酸性炎症和血管功能障碍，并将决定新的早期 BPD的生物标志物。 候选人已经组建了一个具有互补智力的研究咨询委员会， 内容专业知识，可以指导他在整个颁奖期间。此外，候选人将获得教学 教育，以获得必要的知识，最终授予MSPH学位。通过这一综合 指导和教学计划，候选人将获得技能和专业知识，肺部炎症，微生物 致病机理，microRNA和外泌体分子生物学，微生物组测序，生物信息学，技能 无菌小鼠模型的操作和研究伦理。这将使他能够发展一个 独立研究计划，并获得NIH R 01资助。