The function of chemotactic signal transduction during colonization and disease

趋化信号转导在定植和疾病过程中的功能

• 批准号: 9793029
• 负责人: Karen M Ottemann
• 金额: $ 4.36万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-12-01 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9793029
• 关键词: Address; Affect; Affinity; Anti-Bacterial Agents; Antibiotics; Arginine; Australia; Bacteria; Bacterial Infections; Behavior; Binding; Biological Assay; Biology; Cancer Etiology; Cell Communication; Cells; Cessation of life; Chemoreceptors; Chemotactic Factors; Chemotaxis; Collaborations; Cues; Data; Defect; Diffusion; Disease; Disease Outcome; Frequencies; Fumarates; Gastric Glands; Gene Expression; Genes; Goals; Health; Helicobacter Infections; Helicobacter pylori; In Vitro; Incidence; Individual; Infection; Inflammation; Inflammatory; Inflammatory Response; Knowledge; Lead; Ligand Binding; Ligands; Measurement; Mediating; Mission; Modeling; Molecular; Mus; Nature; Nutrient; Organism; Organoids; Outcome; Pathogenesis; Pathogenicity; Pathogenicity Island; Phenotype; Play; Process; Property; Proteins; Public Health; Reagent; Research; Role; Route; Series; Signal Transduction; Signaling Protein; Stomach; Stomach Diseases; Surface Plasmon Resonance; System; Testing; Therapeutic; Thiamine; Ulcer; United States; United States National Institutes of Health; Universities; Work; base; burden of illness; combat; disease phenotype; experimental study; host-microbe interactions; in vivo; innovation; insight; malignant stomach neoplasm; member; metabolomics; microorganism; mouse model; mutant; neutrophil; new therapeutic target; novel; novel therapeutics; overexpression; pathogen; prevent; receptor; response; small molecule; small molecule libraries; tool

Our proposed research focuses on defining the mechanism of action of the TlpA and TlpB chemoreceptors that play fundamental roles in pathogenesis of the ulcer-causing bacterium Helicobacter pylori. These receptors modulate H. pylori-induced inflammation. There is a fundamental gap, however in our understanding of the TlpA and TlpB signals, of how these receptors sense their ligands, how they perform signal transduction, and how they promote pathogenesis. Continued existence of this gap prevents us from gaining a full understanding of H. pylori's pathogenic mechanisms and, in the long term, thwarting these processes to enable the creation of new drugs against H. pylori-related disease. Millions of people worldwide and in the U.S. are infected by H. pylori and suffer from its associated diseases—ulcers and gastric cancer. Gastric cancer is the second cause of cancer deaths worldwide. H. pylori is here to stay based on recent studies that show H. pylori incidence has stabilized in the developed world. Furthermore, current therapies to cure H. pylori infection fail with unaccepta- ble frequency, e.g., recent estimates in the United States have found that 20-25% of infected individuals are not cured by the current therapeutic regime. New drug targets are desperately needed. The specific objective of this application is to dissect TlpA and TlpB signal transduction and its role in gastric disease. Our central hy- pothesis is that TlpA and TlpB transduce information from specific ligands to affect bacterial-host interactions, proinflammatory gene expression, and in turn, host inflammation. Our hypothesis has been formulated from preliminary data using small molecule arrays to identify specific new ligands of TlpA, developing key reagents for analyzing TlpA and TlpB signal transduction, defining the roles of TlpA and TlpB in inflammation, and de- veloping new assays for analyzing H. pylori in the gastric setting. In Aim 1, we determine the mechanism of TlpA and TlpB ligand binding and signal transduction, using in vitro binding assays with purified protein, in vitro chemotaxis signal transduction assays, and intact H. pylori chemotaxis assays. In Aim 2, we determine the mechanism by which TlpA and TlpB modulate inflammation, analyzing the nature of the inflammatory response provoked by tlpA or tlpB mutants. In addition, we will explore the possibility that TlpA and TlpB mutants have abnormal host cell interactions and distribution. Lastly, we investigate the hypothesis that host parameters change of the course of an infection using metabolomics to analyze nutrient distribution in the stomach and state-of-the art organoid culture systems. The proposed research is innovative in that it will create new knowledge about the functions of chemotaxis during bacterial pathogenesis, and it its use of state of the art approaches. The proposed research is significant because it addresses chemotaxis, a fundamental property important to many pathogens, and focuses specifically on a bacterium that causes rampant disease but for which we are losing antibiotic efficacy. The long-term outcomes generated by this research are likely to provide insights that will enable creation of new drugs against H. pylori-related disease.

我们提出的研究重点是定义 TlpA 和 TlpB 化学感受器的作用机制， 这些受体在引起溃疡的细菌幽门螺杆菌的发病机制中发挥着重要作用。 然而，我们对幽门螺杆菌引起的炎症的理解存在根本性的差距。 TlpA 和 TlpB 信号，这些受体如何感知其配体，如何执行信号转导，以及 这种差距的持续存在使我们无法充分了解它们如何促进发病机制。 幽门螺杆菌的致病机制，并从长远来看，阻止这些过程以实现创造 对抗幽门螺杆菌相关疾病的新药。全世界和美国有数百万人感染幽门螺杆菌。 幽门螺杆菌并患有其相关疾病——溃疡和胃癌是第二个原因。 最近的研究表明，幽门螺杆菌的发病率已在全球范围内保持不变。 此外，目前治疗幽门螺杆菌感染的疗法因不可接受的情况而失败。 ble 频率，例如，最近在美国的估计发现 20-25% 的感染者 目前的治疗方案无法治愈，迫切需要新的药物靶点。 本应用的目的是剖析 TlpA 和 TlpB 信号转导及其在胃疾病中的作用。 假设 TlpA 和 TlpB 转导来自特定配体的信息以影响细菌-宿主相互作用， 促炎症基因表达，进而影响宿主炎症，我们的假设是根据这一点制定的。 使用小分子阵列识别 TlpA 特异性新配体的初步数据，开发关键试剂 用于分析 TlpA 和 TlpB 信号转导，定义 TlpA 和 TlpB 在炎症中的作用，以及 开发用于分析胃环境中幽门螺杆菌的新方法 在目标 1 中，我们确定了其机制。 TlpA 和 TlpB 配体结合和信号转导，使用纯化蛋白的体外结合测定，在体外 在目标 2 中，我们确定了趋化信号转导测定和完整幽门螺杆菌趋化测定。 TlpA和TlpB调节炎症的机制，分析炎症反应的性质 由 tlpA 或 tlpB 突变体引起此外，我们将探讨 TlpA 和 TlpB 突变体引起的可能性。 最后，我们研究了宿主参数的假设。 使用代谢组学分析感染过程的变化，分析胃中的营养分布和 所提出的研究具有创新性，因为它将创造新的类器官培养系统。 关于细菌发病机制中趋化性功能的知识及其对现有技术的利用 拟议的研究意义重大，因为它解决了趋化性这一基本特性。 对许多病原体很重要，并特别关注引起猖獗疾病的细菌，但对于 这项研究可能会提供我们正在失去抗生素功效的长期结果。 这些见解将有助于开发针对幽门螺杆菌相关疾病的新药。