Targeted discovery of antibiotics from cave bacteria

从洞穴细菌中有针对性地发现抗生素

基本信息

批准号：
9107281
负责人：
BRADLEY S MOORE
金额：
$ 71.5万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-03-01 至 2020-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9107281
关键词：
Acinetobacter Anabolism Anti-Bacterial Agents Antibiotic Therapy Antibiotics Azithromycin Bacteria Bioinformatics Categories Cells Chemical Structure Cloning Collection Complex Crude Extracts Data Drug resistance Escherichia coli Fluorescence Microscopy Gene Cluster Genes Genome Genomics Goals Gram-Negative Bacteria Growth Hour Individual Label Laboratories Location Mass Spectrum Analysis Measures Membrane Methods Microbe Mining Multi-Drug Resistance Multidrug-resistant Acinetobacter Natural Products New Mexico Organism Orphan Pathway interactions Permeability Pharmacotherapy Prevalence Probability Production Pseudomonas Replication-Associated Process Research Resistance Ribosomal RNA Staphylococcus aureus System Techniques Work base cell envelope cellular targeting combat combinatorial culture plates genome sequencing interest microbial microorganism novel novel strategies overexpression pathogen programs public health relevance resistance gene screening

项目摘要

DESCRIPTION (provided by applicant): The increasing prevalence of bacterial pathogens that are resistant to most of the clinically approved antibiotics is an alarming situation that has spurred renewed interest in antibiotic discovery programs. Since most antibiotics are derived from natural products produced by microorganisms, there is now intense interest in using new methods to screen genetically and chemically diverse collections of bacteria. However, identifying new molecules from bacterial extracts is confounded by the overwhelming presence of previously identified molecules as well as the fact that most of the biosynthetic potential of a organism is typically not expressed under laboratory growth conditions. We have characterized a unique collection of microbes obtained from deep within four different caves of New Mexico. Since these bacteria were isolated from remote, underexplored locations that are only just beginning to be mined for antibiotics, there is an increased probability of identifying molecules with unique chemical structures and new modes of action. The goal of this project is to use two new powerful platforms to identify and purify molecules active against multidrug resistant (MDR) bacteria from this diverse collection of cave bacteria. First, we will use our recently developed bacterial cytological profiling (BCP) approach to identify natural products with antibacterial activities in crude organic extracts or directly on plates. BCP uses quantitative fluorescence microscopy to measure the effects of antibiotic treatment on individual cells. Antibiotics that target different cellular pathways and different steps within a pathway generate unique cytological profiles, allowing identification of the likely cellular target of newly isolated compounds in a few hours. BCP works in complex crude extracts and subsequent fractions, allowing it to be used to guide natural product purification. We will sequence strains producing antibiotics and then use target directed genome mining (TDGM) and heterologous biosynthetic gene cluster (BGC) overproduction to identify novel antibacterial producing BGCs. Heterologous overexpression of normally silent BGCs will allow us to identify molecules missed by traditional screening.

描述（通过应用程序提供）：对大多数临床认可的抗生素具有抗性的细菌病原体的患病率的增加是一种令人震惊的情况激发了对抗生素发现计划的新兴趣。由于大多数抗生素是从微生物产生的天然产物中得出的，因此现在对使用新方法筛查一般和化学多样的细菌集合有浓厚的兴趣。然而，从细菌提取物中鉴定出新的分子是由于先前鉴定的分子的压倒性存在而混淆的，以及在实验室生长条件下通常不会表达生物体的大多数生物合成潜力的事实。我们已经表征了来自新墨西哥州四个不同洞穴中深处获得的独特的微生物集合。由于这些细菌是从刚刚开始用于抗生素的遥远的，未充分膨胀的位置分离出来的，因此鉴定具有独特化学结构和新作用模式的分子的可能性增加了。该项目的目的是使用两个新的强大平台来识别和净化分子，以抵抗这种不同的洞穴细菌收集的耐多药（MDR）细菌。首先，我们将使用我们最近开发的细菌细胞学分析（BCP）方法来鉴定有机提取物中具有抗菌活性或直接在板块上的抗菌活性的天然产物。 BCP使用定量荧光显微镜来测量抗生素处理对单个细胞的影响。靶向不同细胞途径和途径内不同步骤的抗生素会产生独特的细胞学特征，从而在几个小时内鉴定新分离化合物的可能细胞靶标。 BCP以复杂的粗提取物和随后的分数工作，从而使其可用于指导天然产品纯化。我们将对产生抗生素的菌株进行序列菌株，然后使用靶标基因组挖掘（TDGM）和异源生物合成基因簇（BGC）过量生产来鉴定新型的抗菌产生BGC。通常沉默的BGC的异源过表达将使我们能够识别传统筛查错过的分子。