Matrix-associated proteins in biofilm tolerance: Psl and ecotin

生物膜耐受性中的基质相关蛋白：Psl 和 Ecotin

• 批准号: 9011919
• 负责人: Boo Shan Tseng
• 金额: $ 16.16万
• 依托单位: UNIVERSITY OF NEVADA LAS VEGAS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-07 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9011919
• 关键词: Acute; Advisory Committees; Animal Model; Applications Grants; Artificial Heart; Award; Bacteria; Binding; Biology; Biomedical Research; Burn injury; Career Transition Award; Catheters; Cells; Chronic; Chronic Obstructive Airway Disease; Complement; Consultations; Contact Lenses; Cornea; Data; Development; Diabetic mouse; Ear; Extracellular Matrix; Faculty; Future; Goals; Heart Valves; Homologous Gene; Immunoblotting; Immunofluorescence Immunologic; In Vitro; Infection; Job Application; Knowledge; Length; Literature; Lung; Medical Device; Microbial Biofilms; Mission; Modeling; Molecular; Outcome; Pacemakers; Pathogenesis; Pattern; Peptide Hydrolases; Play; Positioning Attribute; Protease Inhibitor; Proteins; Proteomics; Pseudomonas aeruginosa; Public Health; Publishing; Research; Role; Scientist; Serine Proteinase Inhibitors; Surface; Testing; Therapeutic; United States; United States National Institutes of Health; Urinary tract; Work; antimicrobial; base; burden of illness; career; career development; comparative; cystic fibrosis patients; experience; extracellular; insight; pathogen; prevent; programs; public health relevance; therapeutic development; wound

 DESCRIPTION (provided by applicant): While over $25 billion is spent to treat biofilm-based infections in the United States annually, very little is known about the proteins in the biofilm matrix, which protects the resident bacteria from antimicrobial attack. The long-term goal of this application and the Candidate, Dr. Tseng, is to establish an independent research program centered on understanding the function of extracellular matrix-associated proteins in biofilm formation and antimicrobial tolerance. Towards this goal, the immediate career objective of Dr. Tseng is to obtain an independent faculty position, using the research proposed in this application as the cornerstone of her job applications. The overall research objective of this application is to determine the role of ecotin, a serine protease inhibitor that is a putative matrx-associated protein, in the biofilm matrix. The hypothesis is that by inhibiting proteases, ecotin i the biofilm matrix regulates biofilm formation and protects the biofilm from exogenous proteolytic attack. To test this hypothesis, two specific aims are proposed: 1) to evaluate the role of ecotin in protecting the biofilm bacteria, using a diabetic mouse wound model of chronic biofilm infections (in consultation with Dr. Singh) and an in vitro flow cell biofilm model; and 2) to examine the role of ecotin in biofilm formation via immunofluorescence to examine its matrix localization, via qRT-PCR and immunoblot to determine its biofilm expression pattern, and via comparative proteomics of biofilms to examine its function. These aims are expected to demonstrate that enzymatic activities of matrix-associated proteins are important for biofilm formation and antimicrobial tolerance. In addition, the outcomes of these aims are expected to provide preliminary data for a competitive R01 application. The rationale for this research is that it is expected to yield important new insights into biofilm biology, while also providing the means necessary to establish Dr. Tseng, who is committed to a career in basic biomedical research, as an independent molecular microbiologist. In addition to the research described above, this proposal includes career development activities to complement Dr. Tseng's prior experience, as well as an advisory committee assembled by Dr. Tseng. Dr. Parsek, the postdoctoral advisor of Dr. Tseng, is included in this application as a scientific advisor. He is ideal for this position, s he is an internationally recognized expert in biofilm biology with a strong track record of producing successful independent academic scientists. Finally, this Career Transition Award is expected to aid Dr. Tseng in the transition to independence and provide preliminary data for an R01 application within the two years of this award.

 描述（由申请人提供）：虽然美国每年花费超过250亿美元用于治疗基于生物膜的感染，但对生物膜基质中的蛋白质知之甚少，该蛋白质保护常驻细菌免受抗菌剂攻击。该申请和候选人Tseng博士的长期目标是建立一个独立的研究计划，该计划以了解细胞外基质相关蛋白在生物膜形成和抗菌耐受性中的功能为中心。为了实现这一目标，曾博士的直接职业目标是获得一个独立的教师职位，使用本申请中提出的研究作为她工作申请的基石。本申请的总体研究目标是确定ecotin（一种丝氨酸蛋白酶抑制剂，其是推定的基质相关蛋白）在生物膜基质中的作用。假设是通过抑制蛋白酶，生物膜基质中的ecotin调节生物膜形成并保护生物膜免受外源性蛋白水解攻击。为了验证这一假设，提出了两个具体的目标：1）使用慢性生物膜感染的糖尿病小鼠伤口模型来评估ecotin在保护生物膜细菌中的作用（与Singh博士协商）和体外流动细胞生物膜模型;和2）通过免疫荧光检测ecotin在生物膜形成中的作用以检测其基质定位，通过qRT-PCR和免疫印迹确定其生物膜表达模式，并通过生物膜的比较蛋白质组学检查其功能。这些目标有望证明基质相关蛋白的酶活性对于生物膜形成和抗菌耐受性是重要的。此外，这些目标的结果预计将为R 01的竞争性应用提供初步数据。这项研究的基本原理是， 它有望对生物膜生物学产生重要的新见解，同时也提供了 为了使致力于基础生物医学研究的曾博士成为一名独立的分子微生物学家，有必要建立一个独立的分子微生物学家。除了上述研究外，该提案还包括职业发展活动，以补充曾博士先前的经验，以及由曾博士召集的咨询委员会。曾博士的博士后顾问Parsek博士作为科学顾问包括在本申请中。他是这个职位的理想人选，他是国际公认的生物膜生物学专家，在培养成功的独立学术科学家方面有着良好的记录。最后，这个职业过渡奖预计将帮助曾博士在过渡到独立，并提供初步的数据，为R 01申请在两年内，这一奖项。