The Impact of the Cystic Fibrosis infection environment on biofilm development of nontuberculous mycobacteria

囊性纤维化感染环境对非结核分枝杆菌生物膜发育的影响

• 批准号: 10657135
• 负责人: William H. DePas
• 金额: $ 39.12万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-03 至 2028-02-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657135
• 关键词: Affect; Agar; Air; Anoxia; Antibiotic Resistance; Antibiotic Therapy; Antibiotic susceptibility; Antibiotics; Automobile Driving; Bacteria; Biological Assay; Biological Models; Bioreactors; CCRL2 gene; Carbon; Cell physiology; Cells; Chemicals; Clinical; Communities; Cystic Fibrosis; Cystic Fibrosis sputum; Dangerousness; Data; Development; Disease Progression; Environment; Experimental Designs; Gene Expression; Gene Proteins; Genes; Genetic; Genetic Transcription; Genus Mycobacterium; Glutamine; Goals; Growth; Hibernation; Human; Immune response; In Situ; In Vitro; Infection; Libraries; Link; Mediating; Metabolic; Metabolic Pathway; Microbial Biofilms; Modeling; Molecular; Molecular Target; Mutagenesis; Mycobacterium Infections; Mycobacterium abscessus; Mycobacterium smegmatis; Nitrogen; Outcome; Oxygen; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Persons; Pharmaceutical Preparations; Phenotype; Physicians; Physiologic tolerance; Physiological; Predisposition; Prevalence; Process; Public Health; Regimen; Regulation; Research; Resistance; Ribosomes; Role; Shapes; Signal Transduction; Sputum; Structure; System; Techniques; Testing; Tissues; Translations; Variant; Visualization; Work; antibiotic tolerance; cell community; chronic infection; clinically relevant; combat; cystic fibrosis infection; cystic fibrosis patients; dosage; emerging pathogen; experimental study; fighting; genetic regulatory protein; in vitro Assay; in vivo; innovation; insight; lung pathogen; metabolomics; millimeter; molecular targeted therapies; mutant; non-tuberculosis mycobacteria; novel; novel therapeutic intervention; novel therapeutics; pathogen; pathogenic bacteria; prevent; protein function; sensor; therapeutic development; three-dimensional modeling; trait; treatment choice

Project Summary/Abstract -- DePas The emergence of nontuberculous mycobacteria (NTM) as dangerous, antibiotic resistant pulmonary pathogens is outpacing research into their mechanisms of pathogenesis. Our long-term goal is to characterize NTM in the infection environment and develop new therapeutic approaches aimed at specific in vivo bacterial activities. The objective of this proposal is to directly assess NTM biofilm formation and growth rate in situ and determine how the infection environment impacts these processes. We hypothesize that the spatial and chemical environment of sputum from people with Cystic Fibrosis (CF) sputum supports the formation of antibiotic tolerant, slow-growing NTM biofilms through regulated cellular processes. The rationale for this proposal is that the biofilm state and growth rate of a specific bacterial pathogen can have drastic influences on the efficacy of antibiotics and the host immune response. We will test our central hypothesis with two specific aims: 1) Determine how NTM biofilm formation is regulated by the CF chemical environment and how it impacts antibiotic tolerance and 2) Determine how anoxia-induced dormancy influences physiological tolerance and biofilm formation of NTM. The proposed work will combine three innovative complementary techniques into one coherent strategy for investigating infection-relevant phenotypes such as biofilm formation and dormancy. We will employ a novel tissue clearing/bacterial visualization technique MiPACT-HCR in both aims. In Aim 2, we will also utilize a 3D model of the CF infection environment, the Agar Block Biofilm Assay. We will use a new in vitro aggregation assay that allows us to track and quantify the transition from planktonic cells to biofilms in both Aims. The proposal is significant because it will provide an accurate description of the physiological state of NTM during human infection to inform antibiotic choice and dosage decisions. It is also significant in that it will provide molecular targets for development of anti-biofilm and anti-dormancy strategies against NTM. The expected outcome of this work is a thorough understanding of the structure and prevalence of NTM communities during infection of patients with CF and insight into the mechanistic pathways driving biofilm formation and dormancy. These results will have a positive impact by assisting physicians make more appropriate treatment choices for NTM infections.

项目概要/摘要- DePas 非结核分枝杆菌（NTM）的出现是危险的，抗生素耐药性肺结核， 病原体的发病机制研究速度超过了对病原体发病机制的研究。我们的长期目标是 NTM在感染环境中的作用，并开发针对特定体内细菌的新治疗方法 活动该提案的目的是直接评估NTM生物膜的形成和原位生长速率， 确定感染环境如何影响这些过程。我们假设空间和 囊性纤维化（CF）痰液的化学环境支持 抗生素耐受性，通过调节细胞过程缓慢生长的NTM生物膜。这样做的理由 一种建议是，特定细菌病原体的生物膜状态和生长速率可以对细菌的生长产生巨大影响。 抗生素的功效和宿主的免疫反应。我们将用两个具体的例子来检验我们的中心假设。 目的：1）确定NTM生物膜的形成是如何由CF化学环境调节的，以及它是如何被生物膜所控制的。 2）确定缺氧诱导的休眠如何影响生理耐受性 和NTM的生物膜形成。拟议的工作将结合联合收割机三个创新的互补技术 成为一个连贯的策略，用于研究感染相关的表型，如生物膜形成， 休眠我们将在这两个目标中采用一种新的组织清除/细菌可视化技术MiPACT-HCR。 在目标2中，我们还将利用CF感染环境的3D模型，即琼脂块生物膜测定。我们将 使用一种新的体外聚集试验，使我们能够跟踪和量化从增殖细胞到 生物膜在这两个目标。该提案意义重大，因为它将准确描述 NTM在人类感染期间的生理状态，以告知抗生素选择和剂量决定。也是 其重要性在于它将为抗生物膜和抗休眠策略的开发提供分子靶标 反对NTM。这项工作的预期成果是彻底了解结构和流行情况 CF患者感染期间的NTM社区，并深入了解驱动CF的机制途径 生物膜形成和休眠。这些结果将产生积极的影响，协助医生使更多的 NTM感染的适当治疗选择。