Genetic Determinants of the hVISA Mechanism

hVISA 机制的遗传决定因素

• 批准号: 7430691
• 负责人: JOHN E GUSTAFSON
• 金额: $ 28.17万
• 依托单位: NEW MEXICO STATE UNIVERSITY LAS CRUCES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-15 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7430691
• 关键词: Antibiotic Resistance; Antibiotics; Antimicrobial Resistance; Bioinformatics; Cause of Death; Cessation of life; Class; Clinical; Computer software; Data; Data Set; Databases; Development; Exhibits; Foundations; Funding; Gene Expression Profile; Gene Mutation; Genetic Determinism; Genome; Genomics; Genus staphylococcus; Grant; Growth; Hospitals; Infection; Institution; Instruction; Investments; Journals; Manuscripts; Medical Surveillance; Microarray Analysis; Minority; Mission; Mutation; Nature; Nosocomial Infections; Patients; Pliability; Predisposition; Process; Reporting; Research; Research Activity; Resistance; Score; Sequence Analysis; Source; Staphylococcus aureus; Students; Surface; System; Techniques; Time; Today; Training; Treatment Failure; Underrepresented Minority; United States National Institutes of Health; Vancomycin; base; comparative; design; methicillin resistant Staphylococcus aureus; microbial; novel; pathogen; programs; tool; transcriptomics

DESCRIPTION (provided by applicant): Genetic alterations leading to hetero-vancomycin intermediate (hVISA) expression presently defies characterization. A major problem in hVISA mechanistic studies has been the lack of isogenic vancomycin-susceptible and hVISA, especially related or isogenic real-world clinical strains. We have now characterized a clonal hVISA and vancomycin-susceptible S. aureus clinical strain set. The specific aims of this proposal are to determine: (1) genomic alterations that occur in a clinical hVISA and clones of this strain expressing elevated susceptible vancomycin MICs; (2) transcriptome alterations that occur as a result of hVISA mechanism acquisition and genetic alterations leading to elevated vancomycin susceptible MICs; and (3) transcriptome alterations due to vancomycin induction that occur in hVISA and S. aureus strains expressing elevated vancomycin susceptible MICs. The first aim will be accomplished by either 454-based genomic sequencing of a temporally isolated clonal strain set that includes hVISA and non-hVISA expressing varied low-level and elevated susceptible vancomycin MICs, followed by genome annotation and comparison, or Nimblegene comparative genomic sequencing. During this process the PI will be trained on 454-sequencing as well as the genome annotation pipelines XGI, Alpheus, and the experimental GenVar. The second and third aims will be accomplished by comparing and contrasting the transcriptomes of a clonal set of hVISA, non-hVISA strains expressing elevated susceptible vancomycin MICs and non-hVISA strains expressing low-level susceptible vancomycin MICs, isolated following growth with and without vancomycin. These aims will also be enhanced by applying an investigative transcriptome tool referred to as the Staphylococcus microarray meta-database (SAMMD). During this process the PI will be trained on the current NIAID-PFGRC-TIGR-supported S. aureus microarrays version 4 and microarray analysis software. These research objectives are designed to produce enormous datasets that will make this SCORE-funded PI competitive for traditional NIH funding.

描述（由申请方提供）：导致异源万古霉素中间体（hVISA）表达的遗传改变目前无法表征。hVISA机制研究中的一个主要问题是缺乏等基因的万古霉素敏感菌株和hVISA，特别是相关或等基因的真实世界临床菌株。我们现在已经确定了克隆hVISA和万古霉素敏感的S。金黄色葡萄球菌临床菌株集。本提案的具体目的是确定：（1）临床hVISA和表达万古霉素敏感MIC升高的该菌株克隆中发生的基因组改变;（2）由于hVISA机制获得和遗传改变导致万古霉素敏感MIC升高而发生的转录组改变;和（3）由于hVISA和S.第一个目的将通过对暂时分离的克隆菌株集进行基于454的基因组测序来实现，所述克隆菌株集包括表达不同的低水平和升高的敏感万古霉素MIC的hVISA和非hVISA，然后进行基因组注释和比较，或Nimblegene比较基因组测序。在此过程中，PI将接受454测序以及基因组注释管道XGI、Alpheus和实验性GenVar的培训。第二和第三个目的将通过比较和对比hVISA、表达高敏感万古霉素MIC的非hVISA菌株和表达低水平敏感万古霉素MIC的非hVISA菌株的克隆集的转录组来实现，这些菌株在有和没有万古霉素的情况下生长后分离。这些目标也将通过应用被称为葡萄球菌微阵列元数据库（SAMMD）的调查转录组工具来加强。在此过程中，PI将接受关于当前NIAID-PFGRC-TIGR支持的S的培训。aureus微阵列版本4和微阵列分析软件。这些研究目标旨在产生巨大的数据集，使这个SCORE资助的PI与传统的NIH资助相比具有竞争力。