Subproject 4: Role of Pumps in Resistance, Physiology, and Infection

子项目 4：泵在抵抗、生理和感染中的作用

基本信息

批准号：
10571916
负责人：
David C Hooper
金额：
$ 41.48万
依托单位：
MASSACHUSETTS EYE AND EAR INFIRMARY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10571916
关键词：
Abscess Affect Alleles Anti-Bacterial Agents Antibiotic Resistance Antibiotics Antimicrobial Resistance Binding Binding Sites CD36 gene Cell physiology Cells Ciprofloxacin Clinical Collaborations Communities Complex Daptomycin Development Drug Efflux Effectiveness Enterobacteriaceae Enterococcus Enterococcus faecalis Epithelial Cells Fluorescent Probes Fluoroquinolones Fosfomycin Gastrointestinal tract structure Gene Chips Genes Genetic Transcription Genomics Goals Gram-Negative Bacteria Gram-Positive Bacteria Habitats Heterogeneity Hospitals Human In Vitro Individual Infection Invaded Knock-out Libraries Life Measurement Mediating Medium chain fatty acid Membrane Microbial Physiology Modeling Mothers Multi-Drug Resistance Mus Mutagenesis Mutation Pathogenesis Pathway interactions Pattern Phagolysosome Pharmaceutical Preparations Physiology Plasmids Poison Polymyxins Population Predisposition Proteins Public Health Pump Reporter Resistance Resistance development Rifampin Role Site Skin Skin colonization Staphylococcus aureus Structure-Activity Relationship Subcutaneous abscess TLR2 gene Testing Tetanus Helper Peptide Tetracycline Resistance Tissues Trimethoprim-Sulfamethoxazole Variant Work antimicrobial antimicrobial peptide bactericide cellular imaging effective therapy efflux pump expression vector genetic manipulation gut colonization improved in vivo interdisciplinary approach lipoteichoic acid local drug delivery mouse model mutant novel overexpression pathogen peptide analog programs protein expression protein protein interaction resistance mechanism resistance mutation response structural determinants synergism trafficking transposon sequencing

项目摘要

Summary The longterm goals of the project are to identify the full array of efflux pumps of Staphylococcus aureus and Enterococcus spp that contribute to multiple antimicrobial resistance and to elucidate the determinants of their expression, their role in microbial physiology and their effect on bacterial response to antimicrobials in infection. The work will focus on a structural and functional analysis of the Tet38 efflux pump that enables multidrug resistance, survival in abscesses, and invasion of epithelial cells, the potential contribution of multiple pumps to persister subpopulations in the presence of antimicrobials, expanding the understanding of efflux resistance in enterococci, and evaluating collaboratively novel novel antimicrobial compounds for effects of efflux, tissue tolerability, and in vivo efficacy. There are four specific aims: 1) assess the structural determinants of the S. aureus Tet38 efflux pump in resistance, physiology, and pathogenesis and determine the mechanism of its diverse effects on cellular physiology; 2) explore the conditions promoting and heterogeneity of S. aureus efflux pump expression in single-cell groups; 3) evaluate the role of effux pumps in resistance in enterococci; and 4) evaluate the effect of S. aureus efflux pumps on novel compounds from collaborating projects. The work will utilize genetic manipulation, allelic exchange, and expression vectors in S. aureus and E. faecalis, measurements of gene/protein expression with fluorescent reporters, and established murine models of infection (subcutaneous abscess, intestinal colonization) utilizing a genomically defined strains. The overall goal of the program project is to take a well‐integrated, multi‐disciplinary approach to understanding antibiotic resistance development and efflux within cellular physiology combined with the search for compounds that compromise resistant pathogens by inhibiting novel targets and pathways. This project will add to understanding of resistance mechanisms related to multidrug efflux pumps and provide strains for testing the effect of such pumps on novel compounds active against new targets and pathways. It will also utilize a mammalian model of infection to screen new compound tolerability and in vivo activity.

概括该项目的长期目标是确定葡萄球菌的全套外排泵金黄色葡萄球菌和肠球菌属导致多种抗菌药物耐药性并阐明它们表达的决定因素、它们在微生物生理学中的作用以及它们对细菌的影响这项工作将侧重于结构和功能分析。 Tet38 外排泵可实现多重耐药性、脓肿存活和侵袭上皮细胞的多个泵对持久亚群的潜在贡献抗菌药物的存在，扩大对肠球菌外排耐药性的了解，以及合作评估新型抗菌化合物对外排、组织的影响耐受性和体内功效有四个具体目标：1）评估结构决定因素。金黄色葡萄球菌 Tet38 外排泵的耐药性、生理学和发病机制并确定其对细胞生理的多种影响的机制；2）探索促进的条件；以及单细胞群中金黄色葡萄球菌外排泵表达的异质性 3) 评估作用；外排泵对肠球菌的耐药性；4) 评估金黄色葡萄球菌外排泵的效果这项工作将利用基因操作、等位基因来研究合作项目中的新化合物。金黄色葡萄球菌和粪肠球菌中的交换和表达载体，基因/蛋白质的测量用荧光产生器表达，并建立小鼠感染模型（皮下注射）利用基因组确定的菌株来治疗脓肿、肠道定植）的总体目标。计划项目是采用综合的、多学科的方法来了解抗生素细胞生理学中的耐药性发展和外流与寻找相结合通过抑制新靶点和途径来损害耐药病原体的化合物。该项目将加深对与多药外排泵相关的耐药机制的理解，提供菌株来测试此类泵对新化合物的影响它还将利用哺乳动物感染模型来筛选新化合物。耐受性和体内活性。