Targeting cell separation systems of gram-negative bacteria.

针对革兰氏阴性细菌的细胞分离系统。

• 批准号: 9238648
• 负责人: Thomas G Bernhardt
• 金额: $ 50.85万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9238648
• 关键词: Acinetobacter baumannii; Antibiotic Resistance; Antibiotics; Bacteria; Bacterial Infections; Biogenesis; Biological Models; Cell Membrane Permeability; Cell Separation; Cell Wall; Cell division; Cells; Cellular biology; Chemicals; Chemosensitization; Cleaved cell; Clinical; Complex; Detergents; Development; Drug Targeting; Drug resistance; Enterobacter; Enzymes; Escherichia coli; Foundations; Generations; Genetic; Gram-Negative Bacteria; Growth; Hypersensitivity; Incidence; Infection; Klebsiella pneumonia bacterium; Knowledge; Laboratories; Lead; Logic; Mammalian Cell; Membrane; Methods; Modeling; Molecular Genetics; Morphology; Multi-Drug Resistance; Mutation; Mutation Analysis; Nature; New Agents; Newborn Infant; Organism; Pathway interactions; Peptidoglycan; Permeability; Pharmaceutical Preparations; Phase; Process; Proteobacteria; Pseudomonas aeruginosa; Pump; Resistance; Role; Site; System; Therapeutic; Toxic effect; Toxin; United States; amidase; antimicrobial peptide; bacterial resistance; base; cell envelope; combat; constriction; daughter cell; design; drug sensitivity; human disease; inhibitor/antagonist; killings; mutant; new therapeutic target; novel; novel therapeutics; pathogen; programs; public health relevance; scale up; screening; small molecule; small molecule libraries; therapeutic target; tool

DESCRIPTION (provided by applicant): Gram-negative bacteria are the causative agents of a variety of important human diseases. Successful treatment of infections with these organisms is hampered by the complex cell envelope that surrounds them. This envelope includes a second (outer) membrane layer that is difficult for drugs to penetrate. Gram-negative bacteria thus have a high intrinsic resistance to antibiotics and are completely insensitive to many drugs that are effective against bacteria lacking an outer membrane. In this project, we intend to validate new targets for antibiotics that when inactivated will compromise the permeability barrier of the outer membrane to either kill gram-negative bacteria or eliminate their intrinsic resistance to approved therapeutics. The project is based on our studies of cell division in Escherichia coli. In this model gram-negative bacterium, we recently identified factors required for cell envelope remodeling during division and have developed tools to study them. During division, new cell wall material produced by the cytokinetic machinery must be processed by hydrolytic enzymes for daughter cells to separate. We have discovered that this processing is carried out by enzymes called amidases that are activated by factors with LytM domains. Once the cell wall is processed, a second set of factors called the Tol-Pal system catalyzes the constriction of the outer membrane to complete the division process. Inactivation of either the amidases or the Tol-Pal system in E. coli and other gram-negative bacteria leads to the formation of long chains of cells that cannot separate. These cell chains have been shown to have a compromised outer membrane permeability barrier and are hypersensitive to many drugs. Despite their role in maintaining the outer membrane permeability barrier, the importance of these cell separation systems for drug resistance has not been investigated in problematic gram-negative pathogens. Therefore, in this project, we will validate cell separation systems as potential drug targets in te pathogen Pseudomonas aeruginosa. Molecular genetics will be used to investigate the effect of inactivating these systems on the growth and drug resistance of this organism, and chemical screens will be implemented to identify inhibitors of the process. Although we focus on P. aeruginosa, the systems chosen for targeting are highly conserved in gram-negative bacteria. Thus, our results will be broadly relevant to the development of novel treatments for gram-negative infections.

描述（由申请人提供）：革兰氏阴性菌是多种重要人类疾病的病原体。这些生物感染的成功治疗受到它们周围复杂的细胞包膜的阻碍。这个包膜包括药物难以穿透的第二（外）膜层。因此，革兰氏阴性菌对抗生素具有很高的内在耐药性，对许多对缺乏外膜的细菌有效的药物完全不敏感。在这个项目中，我们打算验证抗生素的新靶点，当灭活时将损害外部的渗透性屏障