Understanding the molecular mechanisms and biological functions of a novel bleach-sensing bacterial receptor in shaping host-associated bacterial populations in response to host inflammation

了解新型漂白剂感应细菌受体在塑造宿主相关细菌群体以响应宿主炎症方面的分子机制和生物学功能

• 批准号: 10217956
• 负责人: Arden Baylink
• 金额: $ 11.74万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-16 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10217956
• 关键词: Animal Model; Animals; Antioxidants; Bacteria; Bacterial Infections; Binding; Biochemical Pathway; Biological; Biological Assay; Biological Process; Biological Testing; Biology; Chemistry; Chemoreceptors; Chemotaxis; Chronic; Colon Carcinoma; Communicable Diseases; Crystallography; Cysteine; Disease; Engineering; Enzyme-Linked Immunosorbent Assay; Enzymes; Escherichia coli; Feedback; Fingers; Gastric Tissue; Gastric ulcer; Genetic Engineering; Goals; Helicobacter pylori; Human; Hypochlorous Acid; Immune; Immune system; In Situ; In Vitro; Infection; Infectious Diseases Research; Inflammation; Inflammation Process; Inflammatory Bowel Diseases; Invaded; Investigation; Knowledge; Learning; Leukocytes; Ligands; Measures; Metals; Microbial Biofilms; Microscopy; Modeling; Molecular; Molecular Conformation; Molecular Structure; Mus; Nutrient; Oregon; Oxidants; Oxidation-Reduction; Oxides; Pathogenicity; Pattern; Peroxidases; Physiological; Play; Point Mutation; Population; Process; Production; Proteins; Reaction; Research; Research Personnel; Role; Salmonella; Salmonella typhimurium; Second Messenger Systems; Shapes; Signal Transduction; Signaling Protein; Site; Source; Spatial Distribution; Stomach; Structure; Sulfhydryl Compounds; Swimming; System; Tertiary Protein Structure; Testing; Therapeutic Uses; Tissues; Training; Ulcerative Colitis; Universities; Virulent; Work; Zebrafish; Zinc; antimicrobial; bacterial community; blind; career development; cell motility; chronic infection; commensal bacteria; diguanylate cyclase; drug discovery; experience; experimental study; gut colonization; imaging genetics; in vivo; insight; malignant stomach neoplasm; microbial; mouse model; neutrophil; novel; oxidation; pathogen; pathogenic bacteria; persistent bacteria; prevent; receptor; response; sensor; zinc-binding protein

PROJECT SUMMARY/ABSTRACT A large body of research has elucidated the role of bleach (HOCl) as a potent antimicrobial produced by the immune system through neutrophil myeloperoxidase, but new findings have provided intriguing instances in which this paradigm is challenged, and some important human bacterial pathogens such as Salmonella Typhimurium and Helicobacter pylori have been shown to actually be attracted to inflamed tissue—where HOCl can reach millimolar concentrations—and can exploit host inflammation processes to colonize and establish persistent infections. The molecular basis for how these bacteria can use motility and chemotaxis to respond to HOCl and reaction products is mostly unknown, and represents an important new direction for infectious disease research with the potential to provide many insights into how pathogenic and commensal bacteria colonize and persist in animal hosts to cause disease. The broad and long-term objective of this project is to learn how host-associated bacteria navigate the inflammation and redox landscape of the gut, which has implications for many human infectious diseases, especially those involving bacterial-promoted chronic inflammation such as stomach cancer, ulcerative colitis, and inflammatory bowel diseases. The key focus of this project is to advance understanding of how host-associated bacteria sense and respond to inflammation by investigating a novel bleach-sensing regulatory module referred to as chemoreceptor zinc-binding (CZB) domains that are conserved broadly in commensal and pathogenic bacteria. Specific Aim 1 will use a reductionist in vitro approach to test if CZB domains function through a conserved mechanism by analyzing residue conformation patterns and determining the reactivity with HOCl of representative CZB proteins from H. pylori, S. Typhimurium, and E. coli. Specific Aim 2 will perform the first tests of the biological roles of CZB domains in bacterial infections with a novel ELISA chemotaxis assay and infections in zebrafish and mouse model organisms. Important for the Career Development of Dr. Arden Perkins, this project will provide him comprehensive new training in using animal models, which he has never done, as well as expertise in microscopy and live imaging, and genetic engineering of bacteria. This training will build on Dr. Perkins’ prior experience in protein crystallography, drug discovery, and redox biology to make him a competent investigator at the interface of in vitro and in vivo work, and enable him to pursue investigations into the functions of key proteins involved in bacterial infections at the molecular and biological levels. Dr. Perkins will be trained at University of Oregon in the zebrafish system, a high throughput model in which he will study bacterial dynamics and responses to neutrophils in situ, and collaborators Dr. Manuel Amieva at Stanford and Dr. Andreas Baumler at UC Davis will provide expert guidance and support for extending these studies to mouse models of infection of H. pylori and S. Typhimurium, respectively.

项目摘要/摘要 大量研究阐明了漂白剂（HOCL）的作用，是由 通过嗜中性粒细胞髓过氧化物酶进行免疫系统，但新发现提供了有趣的实例 该范式受到挑战，一些重要的人类细菌病原体（例如沙门氏菌） 幽门螺杆菌和幽门螺杆菌已被证明实际上被炎症组织吸引 可以达到毫米浓度，并可以利用宿主注入过程来定居并建立 持续感染。这些细菌如何使用运动性和趋化性反应的分子基础 HOCL和反应产物大多未知，代表了传染病的重要新方向 研究有可能提供许多有关致病性和共生细菌如何定殖和有多种见解的研究 坚持动物宿主引起疾病。该项目的广泛和长期目标是了解如何 与宿主相关的细菌导航肠道注射和氧化还原景观，这对 许多人类感染性疾病，尤其是那些参与细菌促进的慢性感染的疾病，例如 胃癌，溃疡性结肠炎和炎症性肠病。该项目的重点是进步 了解与宿主相关的细菌如何感知并通过研究新颖 漂白感应调节模块称为化学感受器锌结合（CZB）域 在共生和致病细菌中广泛。特定目标1将使用还原主义的体外方法来测试是否存在 CZB域通过配置机制通过分析居住构象模式和 确定幽门螺杆菌，伤寒链虫和大肠杆菌的代表性CZB蛋白的反应性。 具体目标2将对CZB结构域在细菌感染中的生物学作用进行首次测试。 斑马鱼和小鼠模型生物中的新型ELISA趋化测定和感染。 对于Arden Perkins博士的职业发展至关重要，该项目将为他提供全面的新 他从未做过的动物模型以及显微镜和现场成像方面的专业知识的培训， 和细菌的基因工程。这项培训将基于珀金斯博士先前在蛋白质方面的经验 晶体学，药物发现和氧化还原生物学，使他成为In界面的合格研究者 体外和体内工作，使他能够对涉及的关键蛋白质的功能进行调查 分子和生物学水平的细菌感染。珀金斯博士将在俄勒冈大学的培训 斑马鱼系统，这是一种高吞吐量模型，他将研究细菌动力学和对 中性粒细胞原位，以及斯坦福大学的Manuel Amieva博士和加州大学戴维斯分校的Andreas Baumler博士 提供专家指导和支持，以将这些研究扩展到幽门螺杆菌感染的小鼠模型和 鼠伤寒S.。

项目成果

Rural exclusion from science and academia.

• DOI: 10.1016/j.tim.2021.06.012
• 发表时间: 2021-11
• 期刊: Trends in microbiology
• 影响因子: 15.9
• 作者: O'Neal L;Perkins A
• 通讯作者: Perkins A

A Bacterial Inflammation Sensor Regulates c-di-GMP Signaling, Adhesion, and Biofilm Formation.

• DOI: 10.1128/mbio.00173-21
• 发表时间: 2021-06-29
• 期刊: mBio
• 影响因子: 6.4
• 作者: Perkins A;Tudorica DA;Teixeira RD;Schirmer T;Zumwalt L;Ogba OM;Cassidy CK;Stansfeld PJ;Guillemin K
• 通讯作者: Guillemin K