Elucidating how CRISPR-Cas Modulates the Spread of S. aureus Pathogenicity Islands

阐明 CRISPR-Cas 如何调节金黄色葡萄球菌致病岛的传播

• 批准号: 10473519
• 负责人: Dalton Van Banh
• 金额: $ 5.18万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-06 至 2025-08-05
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10473519
• 关键词: Affect; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Automobile Driving; Bacteria; Bacterial Chromosomes; Bacterial Infections; Bacteriophages; Biological; Biological Assay; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Community-Acquired Infections; Complex; Data; Development; Disease; Doctor of Philosophy; Elements; Enterotoxins; Evolution; Excision; Fellowship; Foundations; GTP-Binding Protein alpha Subunits, Gs; Genes; Genetic; Genetic Transcription; Genome; Genus staphylococcus; Goals; Horizontal Gene Transfer; Immunity; Immunization; Infection; Invaded; Laboratories; Larva; Life Cycle Stages; Measures; Mediating; Medical; Mobile Genetic Elements; Modeling; Molecular; Molecular Genetics; Multi-Drug Resistance; Multiple Bacterial Drug Resistance; Nosocomial Infections; Pathogenesis; Pathogenicity Island; Physicians; Plasmids; Population; Principal Investigator; RNA; Research; Ribonucleases; Scientist; Staphylococcus aureus; Superantigens; System; Testing; Tetracycline Resistance; Therapeutic; Toxic Shock Syndrome; Toxin; Training; Virulence; Virulence Factors; Virulent; Work; adaptive immunity; antimicrobial; base; career; colonization resistance; commensal microbes; design; experimental study; genetic approach; global health; in vivo; insight; multi-drug resistant pathogen; mutant; novel therapeutic intervention; nuclease; particle; pathogen; prevent; programs; reproductive; resistance gene; staphylococcal enterotoxin; targeted treatment; transmission process; Affect; Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Automobile Driving; Bacteria; Bacterial Chromosomes; Bacterial Infections; Bacteriophages; Biological; Biological Assay; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Community-Acquired Infections; Complex; Data; Development; Disease; Doctor of Philosophy; Elements; Enterotoxins; Evolution; Excision; Fellowship; Foundations; GTP-Binding Protein alpha Subunits, Gs; Genes; Genetic; Genetic Transcription; Genome; Genus staphylococcus; Goals; Horizontal Gene Transfer; Immunity; Immunization; Infection; Invaded; Laboratories; Larva; Life Cycle Stages; Measures; Mediating; Medical; Mobile Genetic Elements; Modeling; Molecular; Molecular Genetics; Multi-Drug Resistance; Multiple Bacterial Drug Resistance; Nosocomial Infections; Pathogenesis; Pathogenicity Island; Physicians; Plasmids; Population; Principal Investigator; RNA; Research; Ribonucleases; Scientist; Staphylococcus aureus; Superantigens; System; Testing; Tetracycline Resistance; Therapeutic; Toxic Shock Syndrome; Toxin; Training; Virulence; Virulence Factors; Virulent; Work; adaptive immunity; antimicrobial; base; career; colonization resistance; commensal microbes; design; experimental study; genetic approach; global health; in vivo; insight; multi-drug resistant pathogen; mutant; novel therapeutic intervention; nuclease; particle; pathogen; prevent; programs; reproductive; resistance gene; staphylococcal enterotoxin; targeted treatment; transmission process

PROJECT SUMMARY/ABSTRACT Staphylococcus aureus is a major cause of both community-acquired and nosocomial infections, which have become increasingly challenging to treat due to the widespread evolution of antimicrobial resistance. There is a critical need for the development of alternative therapeutic approaches against multidrug-resistant bacteria that also spare the protective commensal microbiota, which often provide colonization resistance against pathogens. Bacterial disease is driven by S. aureus toxins and other virulence factors, which are mainly encoded by mobile genetic elements (MGEs). In particular, numerous enterotoxins and the superantigen toxin causing Toxic Shock Syndrome are all carried by a class of MGEs called the S. aureus Pathogenicity Islands (SaPIs), which spread between bacteria by hijacking the reproductive machinery of bacteriophages. Staphylococci also possess CRISPR-Cas systems, which provide adaptive immunity by blocking invading MGEs like phages and plasmids. In this proposal, building upon preliminary data, I will test the central hypothesis that CRISPR-Cas systems also prevent the transmission of SaPI elements and their associated virulence genes. In Aim 1, I will define the complex tripartite interplay between staphylococcal CRISPR systems, SaPIs, and their helper phages using various molecular and genetic approaches. In Aim 2, I will investigate the mechanisms by which SaPIs manage to overcome CRISPR-mediated restriction and disseminate throughout bacterial populations. I anticipate that these studies will elucidate both the molecular basis and biological consequences for CRISPR-SaPI interactions. In Aim 3, I will evaluate whether CRISPR can be used to selectively kill SaPI-harboring S. aureus and establish a proof-of-concept for CRISPR-based antimicrobials directed against virulence-encoding MGEs. The proposed experiments will contribute to the long-term goal of designing alternative therapeutic approaches in an effort to overcome the shortcomings of antibiotics in treating multidrug-resistant infections. This fellowship will support my training in the Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD Program, including my doctoral work in the laboratory of Dr. Luciano Marraffini at Rockefeller and the remainder of my medical training at Weill Cornell. The training plan outlined in this fellowship project is designed to optimally prepare me for a research career as an independent principal investigator and physician-scientist.

项目摘要/摘要 金黄色葡萄球菌是社区获得性感染和医院感染的主要原因， 由于抗菌素耐药性的广泛演变，治疗越来越具有挑战性。有一个 针对多种耐药细菌开发替代治疗方法的批判性需求 还要避免保护性共生微生物群，该菌群通常会对病原体提供定殖的抗性。 细菌疾病是由金黄色葡萄球菌毒素和其他毒力因素驱动的，这些因素主要由移动 遗传因素（MGE）。特别是，许多肠毒素和超抗原毒素引起有毒休克 综合征全部由一类称为金黄色葡萄球菌致病岛（SAPI）的MGE携带 通过劫持噬菌体的生殖机械之间的细菌之间。葡萄球菌也拥有 CRISPR-CAS系统，通过阻止噬菌体和质粒等入侵的MGE来提供自适应免疫。 在此提案的基于初步数据的基础上，我将测试CRISPR-CAS系统的中心假设 防止SAPI元素的传播及其相关的毒力基因。在AIM 1中，我将定义 使用葡萄球菌CRISPR系统，SAPI及其辅助噬菌体之间的复杂三方相互作用 各种分子和遗传方法。在AIM 2中，我将调查SAPIS管理的机制 为了克服CRISPR介导的限制并在细菌种群中传播。我预料到了 这些研究将阐明CRISPR-SAPI相互作用的分子基础和生物学后果。 在AIM 3中，我将评估CRISPR是否可以选择性地杀死Sapi-Harboring S. Aureus并建立 基于CRISPR的抗菌剂的概念验证，针对毒力编码的MGE。提议 实验将有助于设计替代治疗方法的长期目标，以便 克服抗生素在治疗多药耐药感染中的缺点。这个奖学金将支持 我在Weill Cornell/Rockefeller/Sloan Kettering三机构MD-PHD计划中进行的培训，包括我 洛克菲勒（Rockefeller）Luciano Marraffini博士和我的其余医学培训的博士学位 在威尔·康奈尔（Weill Cornell）。该奖学金项目中概述的培训计划旨在为我做好准备 研究职业是独立的首席研究员和医师科学家。