Functionality of CdrA and its processed forms within Pseudomonas aeruginosa biofilms

CdrA 的功能及其在铜绿假单胞菌生物膜中的加工形式

• 批准号: 10661570
• 负责人: Courtney Reichhardt
• 金额: $ 24.9万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10661570
• 关键词: Address; Antibiotic Therapy; Antibiotics; Architecture; Bacteria; Bacterial Adhesins; Binding; Biochemical; Biological; Biological Assay; Burn injury; Catheters; Chronic; Clinical; Complement; Contact Lenses; DNA; Data; Development Plans; Extracellular Matrix; Faculty; Foundations; Funding; Future; Goals; Growth; Host Defense; Host Defense Mechanism; Immune response; Immune system; Infection; Integration Host Factors; International; Job Application; Laser Scanning Confocal Microscopy; Mechanics; Mediating; Mentors; Mentorship; Methods; Microbe; Microbial Biofilms; Microbiology; Microscopy; Mission; Modeling; Mucins; Nature; Outcome; Peptide Hydrolases; Play; Positioning Attribute; Prevention; Process; Proteins; Proteolysis; Proteolytic Processing; Pseudomonas aeruginosa; Public Health; Research; Resistance; Scientist; Structure; Testing; Tissues; Training; United States National Institutes of Health; Work; antimicrobial; assault; biophysical chemistry; career; career development; chronic infection; crosslink; cystic fibrosis patients; effective therapy; environmental change; experience; extracellular; flexibility; improved; interdisciplinary approach; interest; intermolecular interaction; microbial community; multidisciplinary; pathogen; professor; programs; research faculty; scaffold; solid state nuclear magnetic resonance; tool; urinary; wound

PROJECT SUMMARY/ABSTRACT Microbial communities called biofilms are hallmarks of chronic infections. Pseudomonas aeruginosa is a model for biofilm study, and also is a pathogen that causes biofilm infections including of burns, wounds, urinary catheters, contact lenses, and the airways of patients with cystic fibrosis (CF). Biofilm microbes are encased in a mesh-like extracellular matrix that helps bacteria to evade host defenses and protects against external assaults such as antimicrobial treatment. A better understanding of biofilm matrix assembly is necessary to improve prevention and treatment of biofilm-involved infections. The long-term goal of this application and the candidate, Dr. Reichhardt, is to establish an independent research program focused on the nature and functional impact of intermolecular interactions in biofilm. Towards this goal, the immediate career objective of Dr. Reichhardt is to obtain an independent faculty position using the proposed research as the foundation of her job applications. The overall research objective of this application is to investigate the flexibility and control of matrix interactions of the key P. aeruginosa protein CdrA. Based on preliminary data, the hypothesis is that the biofilm functionality of CdrA can be expanded through enzymatic processing and binding to host factors, impacting biofilm stability. To test this hypothesis, two specific aims are proposed and will be investigated using Dr. Reichhardt’s unique multidisciplinary training in biophysical chemistry and microbiology. Aim 1 will elucidate the biofilm functionality of CdrA and its processed forms within P. aeruginosa biofilms including those found in chronic infections. Aim 2 will examine the ability of CdrA to integrate host material into biofilms and determine if these host factor- integrated biofilms are more protected against antimicrobials and host immune responses. These aims are expected to transform the established view of biofilm matrices as static structures that are formed by only self- produced biomolecules. This improved understanding will advance biofilm microbiology and aid the creation of effective treatments for chronic infections. This proposal includes a career development plan to complement Dr. Reichhardt’s prior experience so that she can successfully transition to an independent research faculty position. Dr. Reichhardt has assembled a multidisciplinary mentorship committee to help her achieve her scientific and career development goals. Professor Parsek is an ideal mentor for Dr. Reichhardt since he is internationally recognized as a leader in biofilm microbiology, and he has a strong track record producing successful independent academic scientists, including several with multidisciplinary backgrounds. The proposed research is distinct from that of Professor Parsek, and the results of this project will transition with Dr. Reichhardt to her future independent research program. Additionally, results generated from the proposed research aims are expected to help Dr. Reichhardt successfully compete for R01 funding.

项目摘要/摘要 被称为生物膜的微生物群落是慢性感染的标志。铜绿假单胞菌是一个典范 用于生物被膜研究，也是引起生物被膜感染的病原体，包括烧伤、伤口、尿路 囊性纤维化(CF)患者的导管、隐形眼镜和呼吸道。生物膜微生物被包裹在 一种网状细胞外基质，帮助细菌逃避宿主防御并抵御外部攻击 比如抗菌治疗。有必要更好地了解生物膜基质组装，以提高 预防和治疗生物被膜感染。这份申请和候选人的长期目标， Reichhardt博士将建立一个独立的研究计划，专注于脑血管疾病的性质和功能影响 生物膜中的分子间相互作用。为了实现这一目标，赖克哈特博士的直接职业目标是 利用所提议的研究作为她工作申请的基础，获得一个独立的教员职位。 该应用程序的总体研究目标是研究基质相互作用的灵活性和可控性 铜绿假单胞菌的关键蛋白CDRA。根据初步数据，假设生物膜的功能 CDRA可通过酶处理和与宿主因子结合而扩增，影响生物膜的稳定性。 为了验证这一假设，提出了两个具体的目标，并将使用Reichhardt博士独特的 生物物理化学和微生物学方面的多学科培训。目标1将阐明生物膜的功能 CDRA及其在铜绿假单胞菌生物膜内的加工形式，包括在慢性感染中发现的那些。目标2 将检查CDRA将宿主材料整合到生物膜中的能力，并确定这些宿主因素是否- 集成的生物膜可以更好地抵御抗菌剂和宿主免疫反应。这些目标是 预计将改变生物膜基质作为静态结构的既定观点，这些静态结构仅由自身形成 产生了生物分子。这一更好的理解将促进生物膜微生物学的发展，并有助于创造 治疗慢性感染的有效方法。该提案包括一项职业发展计划，以补充Dr。 Reichhardt以前的经验，使她可以成功地过渡到一个独立的研究教授的职位。 Reichhardt博士已经组建了一个多学科指导委员会来帮助她实现她的科学和 职业发展目标。Parsek教授是Reichhardt博士的理想导师，因为他在国际上 被公认为生物膜微生物学的领导者，他有很好的记录，生产成功 独立的学术科学家，包括几个具有多学科背景的人。拟议的研究 不同于Parsek教授的，这个项目的结果将从Reichhardt博士过渡到她 未来的独立研究计划。此外，建议的研究目标所产生的结果如下 预计将帮助Reichhardt博士成功竞争R01资金。