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.

项目总结/摘要 大量的研究已经阐明了漂白剂（HOCl）作为由细菌产生的有效抗菌剂的作用。 免疫系统通过中性粒细胞髓过氧化物酶，但新的发现提供了有趣的例子， 这种模式受到挑战，一些重要的人类细菌病原体，如沙门氏菌， 鼠伤寒和幽门螺杆菌已被证明实际上是吸引到发炎的组织-其中HOCl 可以达到毫摩尔浓度，并可以利用宿主炎症过程来殖民和建立 持续感染。这些细菌如何利用运动性和趋化性来响应 HOCl和反应产物大多是未知的，代表了感染性疾病的重要新方向 研究有可能提供许多关于致病菌和寄生菌如何定植的见解， 在动物宿主中持续存在并引起疾病。这个项目的广泛和长期目标是学习如何 宿主相关细菌在肠道的炎症和氧化还原环境中导航，这对 许多人类感染性疾病，特别是那些涉及细菌促进的慢性炎症， 胃癌、溃疡性结肠炎和炎症性肠病。该项目的重点是推进 了解宿主相关细菌如何通过研究一种新的 漂白敏感调节模块称为化学受体锌结合（CZB）结构域，其是保守的 广泛存在于肠道和致病菌中。具体目标1将使用体外还原方法来测试， CZB结构域通过分析残基构象模式的保守机制发挥功能， 测定来自H. pylori、S. Typhimurium和E.杆菌 Specific Aim 2将首次测试CZB结构域在细菌感染中的生物学作用， 新的ELISA趋化性测定和感染斑马鱼和小鼠模式生物。 重要的是博士的职业发展雅顿帕金斯，这个项目将为他提供全面的新的 他接受过使用动物模型的训练，这是他从未做过的，以及显微镜和实时成像的专业知识， 和细菌的基因工程。这项培训将建立在珀金斯博士先前在蛋白质方面的经验基础上 结晶学，药物发现和氧化还原生物学，使他成为一个称职的研究人员在接口的 体外和体内工作，使他能够继续研究参与的关键蛋白质的功能， 在分子和生物水平上的细菌感染。帕金斯博士将在俄勒冈州大学接受培训， 斑马鱼系统，一个高通量模型，他将研究细菌动力学和对 中性粒细胞原位，合作者Manuel Amieva博士在斯坦福大学和Andreas Baumler博士在加州大学戴维斯分校将 为将这些研究扩展到H.与幽门 S.分别为鼠伤寒。

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