Uncovering aryl polyene biology to identify new drug targets in Gram-negative bacterial pathogens

揭示芳基多烯生物学以确定革兰氏阴性细菌病原体的新药物靶点

• 批准号: 10381676
• 负责人: Jan Claesen
• 金额: $ 40.25万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10381676
• 关键词: Acinetobacter baumannii; Address; Affect; Anabolism; Anti-Bacterial Agents; Antibiotics; Bacteria; Bacterial Antibiotic Resistance; Bacterial Genome; Bacteroidetes; Biochemical; Biological Models; Biology; Carotenoids; Cell membrane; Cells; Cessation of life; Characteristics; Communicable Diseases; Communities; Complex; Contracts; Data; Defense Mechanisms; Development; Drug Targeting; Enterobacter; Enzymes; Excision; Family; Future; Gene Cluster; Genes; Goals; Gram-Negative Bacteria; Healthcare; Immune; Individual; Infection; Link; Mediating; Membrane; Molecular; Molecular Genetics; Pathogenicity; Pathway interactions; Persons; Phagocytes; Pharmaceutical Preparations; Polyenes; Prevention; Production; Proteobacteria; Reactive Oxygen Species; Regulation; Regulator Genes; Regulatory Pathway; Research; Research Personnel; Resistance; Resistance development; Running; Series; Societies; Staphylococcus aureus; Substrate Specificity; Testing; Therapeutic; United States; Uropathogenic E. coli; Virulence; Virulence Factors; analog; cell envelope; combat; cost; forward genetics; genetic analysis; genetic approach; human pathogen; immune clearance; in silico; in vivo; inhibitor; insight; liquid chromatography mass spectrometry; multi-drug resistant pathogen; mutant; new therapeutic target; novel; opportunistic pathogen; pathobiont; pathogen; pathogenic Escherichia coli; pathogenic bacteria; pressure; small molecule; staphyloxanthin; tool

ABSTRACT We are rapidly running out of options to combat infectious diseases in an era that is marked by a rise in antibiotic resistant bacterial pathogens. The discovery rate of novel antibiotics is not keeping up with this pressing need and thus we have to identify alternative drug targets and develop complementary elimination strategies. This is a particularly challenging task for infections caused by Gram-negative bacteria because their outer membrane is impermeable to many compounds. We discovered an aryl polyene (APE) virulence factor, widespread among Gram-negative pathogens, and here propose to characterize its biosynthesis and use it as a target for the development of elimination strategies. APEs are the product of the most abundant family of biosynthetic gene clusters (BGCs) in our global in silico analysis of bacterial genomes. They are present in several multidrug- resistant pathogens, and our preliminary data shows that APEs are esterified to a yet unidentified anchor molecule in the Gram-negative outer membrane. Furthermore, we discovered that APEs serve as virulence factors that protect their producers from host innate immune clearance mechanisms. Our long-term goal is to understand APE biology on a molecular, functional and evolutionary level. The current proposal is centered on the hypothesis that APE biosynthesis and cell envelope localization provide targets for specific antivirulence strategies against Gram-negative pathogens. We will test this by first characterizing the function of individual genes within the APE outer membrane localization pathway and identifying the APE-containing anchor molecule. Next, we will determine the environmental conditions that control regulation of APE expression using a forward genetics approach. Finally, we will develop interference strategies to inhibit APE production, using a targeted inhibitor of the initiation enzyme in the biosynthetic pathway. Our proposed project will yield mechanistic insights into the biology of APEs, as well as identify small molecule tool compounds that can be used to study APE biology and enable alternative pathogen eradication strategies.

抽象的 在一个以抗生素升高为标志的时代，我们迅速无法选择对抗传染病的选择 抗性细菌病原体。新型抗生素的发现率并不能满足这种紧迫的需求 因此，我们必须确定替代药物目标并制定互补的消除策略。这是 革兰氏阴性细菌引起的感染的一项特别具有挑战性的任务，因为其外膜 许多化合物不渗透。我们发现了一个芳基多烯（APE）毒力因子，在 革兰氏阴性病原体，并在这里提出表征其生物合成并将其用作目标 制定消除策略。猿类是最丰富的生物合成基因家族的产物 在我们的全球硅细菌基因组分析中，簇（BGC）。它们存在于几个多饮用 抗性病原体，我们的初步数据表明，将猿酯化为尚未确定的锚 革兰氏阴性外膜中的分子。此外，我们发现猿类是毒力 保护生产者免受宿主先天免疫清除机制的因素。我们的长期目标是 了解分子，功能和进化水平上的猿类生物学。 当前的建议集于以下假设 针对革兰氏阴性病原体的特定抗电力策略的靶标。我们将首先测试 表征猿外膜定位途径中各个基因的功能，并且 识别含猿的锚固分子。接下来，我们将确定环境条件 使用正向遗传学方法控制猿表达的调节。最后，我们将发展干扰 使用生物合成途径中起始酶的靶向抑制剂抑制猿产生的策略。 我们提出的项目将产生对猿类生物学的机械见解，并识别小分子 可用于研究猿类生物学并启用替代性病原体策略的工具化合物。