Evaluating Mycobacterium avium glycopeptidolipids as key factors in the transition from biofilm to macrophages

评估鸟分枝杆菌糖肽脂作为从生物膜向巨噬细胞转变的关键因素

基本信息

批准号：
10582714
负责人：
JEFFREY Scott SCHOREY
金额：
$ 19.56万
依托单位：
UNIVERSITY OF NOTRE DAME
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：
2022-03-02 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10582714
关键词：
Address Anabolism Antibiotic Therapy Antibiotics Bacillus Bacteria Biological Biology Cell surface Cells Chronic lung disease Clinical Data Diagnosis Disease Elderly Engineering Environment Gene Expression Genes Genetic Genetic Transcription Genus Mycobacterium Goals Growth Guidelines Habitats Household Human Immune Individual Infection Invaded Knowledge Life Style Lung Lung diseases Macrophage Macrophage Activation Microbe Microbial Biofilms Modification Mycobacterium avium Mycobacterium avium Complex Mycobacterium avium-intracellulare Infection Mycobacterium tuberculosis Opportunistic Infections Organism Pathogenesis Patients Pattern recognition receptor Phagocytosis Phagosomes Phenotype Play Plumbing Principal Investigator Process Relapse Respiratory Disease Role Slide Soil Source Sputum Structure Surface System Testing Therapeutic United States Variant cell motility common treatment differential expression knockout gene lung injury mutant non-tuberculosis mycobacteria pathogen patient population programs recurrent infection stressor success transcriptome transmission process

项目摘要

Project summary Non-tuberculosis mycobacteria like M. avium are widespread environmental organisms, occurring in many habitats, both natural and engineered (). The ability of M. avium to colonize household plumbing, hot tubs, and garden soils, often places them in close proximity with humans. Most cases of disease caused by M. avium are pulmonary and the patient population includes individuals who are elderly, immune-compromised, or have preexisting lung damage due to an underlying respiratory disease (). Cases of M. avium disease have continued to rise, with cases increasing an estimated 5-10% annually in the United States (). M. avium pulmonary disease is both difficult to diagnose and treat. Once diagnosed, treatment with multiple antibiotics can last 18-24 months, with clinical guidelines indicating continuous treatment for 12 months after the infecting bacteria are no longer detected in patient sputum. Even with this extended treatment course, recurrence of infection is still common. M. avium is an intracellular pathogen with macrophages being the primary host cell. Therefore for the infection to be successful, the mycobacteria must adjust from living in its natural reservoir within a biofilm to invading and replicating in a macrophage. We know very little about how the mycobacteria makes this critical transition. We hypothesize that modifications of glycopeptidolipids, which are major cell surface molecules of M. avium, play a critical role in this transition and evidence showing their importance in biofilm formation and macrophage activation supports this argument. In Specific aim 1 we will test this hypothesis by isolating, from biofilm and macrophages, the different M. avium GPL species and characterize their ability to modulate macrophage function. We will also generate gene knockouts to produce GPL mutants and evaluate the M. avium mutants for biofilm formation and macrophage invasion/survival to define what GPL species are necessary for the observed phenotypes. In aim 2 we will perform a transcriptional analysis of M. avium grown in broth culture, biofilms and macrophages to define expression levels of the different genes involved in producing the various GPLs variants. The transcriptional analysis will have the added benefit of defining more globally, changes in gene expression which occur as the M. avium transitions from growth in biofilm to replication in macrophages.

项目摘要鸟杆菌（M. Avium 发生在许多天然和工程的栖息地中（）。大雄杆菌定居的能力家庭管道，热水浴缸和花园土壤，通常将它们与人类。大多数由大雄性大麻菌引起的疾病病例是肺部和患者人群包括老年人，免疫受损或具有肺部损害的人由于潜在的呼吸系统疾病（）。大雄性疾病的病例继续增加，在美国，每年估计增加5-10％的病例（）。 M. avium肺疾病既难以诊断和治疗。一旦诊断出多次治疗抗生素可以持续18-24个月，并有临床指南表明持续治疗12 在患者痰中不再检测到感染细菌后的几个月。即使这样扩展的治疗过程，感染的复发仍然很常见。大鸟肉是一种细胞内病原体，巨噬细胞是主要宿主细胞。所以为了使感染成功，分枝杆菌必须根据其自然储层生活进行调整在生物膜内入侵和复制巨噬细胞。我们对如何了解分枝杆菌使这种关键过渡。我们假设对糖肽寡脂是大鸟支原体的主要细胞表面分子，在此起着至关重要的作用过渡和证据表明它们在生物膜形成和巨噬细胞中的重要性激活支持此论点。在特定目的1中，我们将通过隔离，从生物膜和巨噬细胞中，不同的Avium GPL物种并表征其能力调节巨噬细胞功能。我们还将生成基因敲除以产生GPL 突变体并评估生物膜形成和巨噬细胞的鸟藻突变体侵袭/生存以定义观察到的表型所需的GPL物种。在 AIM 2我们将对在肉汤培养，生物膜和巨噬细胞定义涉及的不同基因的表达水平各种GPLS变体。转录分析将具有定义更多的额外好处在全球范围内，基因表达的变化是随着鸟分枝杆菌的生长转变而发生的生物膜在巨噬细胞中复制。