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

基本信息

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
项目类别：
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

项目摘要

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.

项目总结/摘要——DePas 非结核分枝杆菌 (NTM) 成为危险的、对抗生素耐药的肺部细菌病原体的研究速度超过了对其发病机制的研究。我们的长期目标是塑造感染环境中的NTM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原位 NTM 生物膜的形成和生长速度，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) 患者痰液的化学环境支持形成通过调节细胞过程产生抗生素耐受性、生长缓慢的 NTM 生物膜。这样做的理由建议认为，特定细菌病原体的生物膜状态和生长速率可能会对抗生素的功效和宿主的免疫反应。我们将用两个具体的方法来检验我们的中心假设目标：1) 确定 NTM 生物膜的形成如何受 CF 化学环境的调节以及它是如何调节的影响抗生素耐受性，2) 确定缺氧诱导的休眠如何影响生理耐受性和NTM生物膜的形成。拟议的工作将结合三种创新的互补技术形成一种连贯的策略来研究感染相关的表型，例如生物膜的形成和休眠。我们将在这两个目标中采用新型组织透明/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测定，使我们能够跟踪和量化从浮游细胞到两个目标中的生物膜。该提案意义重大，因为它将提供对人类感染期间 NTM 的生理状态可为抗生素选择和剂量决策提供信息。这也是重要的是它将为抗生物膜和抗休眠策略的开发提供分子靶点反对NTM。这项工作的预期成果是对结构和流行程度的透彻了解 CF 患者感染过程中 NTM 群落的变化及其驱动机制的洞察生物膜的形成和休眠。这些结果将产生积极影响，帮助医生做出更多努力 NTM 感染的适当治疗选择。