The role of ArsS in gastric infection by Helicobacter pylori

ArsS在幽门螺杆菌胃部感染中的作用

• 批准号: 8864812
• 负责人: George Sachs
• 金额: $ 28.46万
• 依托单位: BRENTWOOD BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8864812
• 关键词: Acclimatization; Acidity; Acids; Affinity; Ammonia; Anti-Infective Agents; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Bacterial Proteins; Binding; Biochemical; Biological; Biology; Campylobacter jejuni; Clarithromycin; Collaborations; Complex; Data; Decarboxylation; Detection; Development; Drug Targeting; Duodenal Ulcer; Gastric Acid; Gastritis; Goals; Helicobacter Infections; Helicobacter pylori; Hydrolysis; Infection; Knowledge; Lead; Libraries; Maintenance; Malignant Neoplasms; Mediating; Membrane; Membrane Protein Traffic; Metronidazole; Microbe; Molecular; Mono-S; Organism; Outcome Study; Peptic Ulcer; Phosphotransferases; Physiology; Population; Preventive; Proteins; Proteome; Proteomics; Proton Pump Inhibitors; Protons; Public Health; Pylorus; Regimen; Regulation; Regulatory Pathway; Research; Resistance; Resources; Role; Signal Pathway; Signal Transduction; Signaling Protein; Specificity; Stomach; Stomach Carcinoma; Stomach Diseases; System; Techniques; Testing; Treatment Protocols; Urea; Urease; Vibrio cholerae; Work; acid stress; bactericide; base; biological adaptation to stress; design; falls; high throughput screening; improved; infectious disease treatment; inhibitor/antagonist; innovation; microbiome; mucosa-associated lymphoid tissue lymphoma; novel; pathogenic Escherichia coli; pathogenic bacteria; periplasm; protein transport; protonation; public health relevance; response; screening; sensor; targeted treatment; trafficking

 DESCRIPTION (provided by applicant): Helicobacter pylori colonizes the normal acid-secreting stomach of about 50% of the world's population. Colonization is associated with gastric disease, including gastritis, peptic and duodenal ulcers, gastric carcinoma, and MALT lymphoma. Although initial eradication rates with triple therapy were successful (~95%), antibiotic resistance has made successful treatment of infection progressively more difficult. Thus, there is a critical need for an antibiotic-free alternative eradication mono-therapy that is . pylori-specific, sparing commensal gut flora or improvement to antibiotic therapy that decreases the duration and complexity of treatment. The long-term goal of this study is to understand how H. pylori responds to and survives gastric acidity and exploit this knowledge to develop novel treatment regimens and/or improve current eradication therapies. The objective here is to determine how the acid-sensitive Two-Component System (TCS), ArsRS, regulates the acid-induced trafficking of urease and its accessory proteins to the inner membrane to form a membrane- bound complex with UreI that is required for gastric infection and acid survival. The rationale for the study is that understanding the mechanisms used by ArsS will allow pharmacological interference resulting in new H. pylori-specific antibiotic-free monotherapy and/or improvement in current eradication regimens. The specific aims are: 1. Elucidate the acid-induced signaling mechanism mediated by the sensor kinase ArsS. Because ArsS is essential for gastric colonization and acid survival, knowing the mechanism of protein trafficking to UreI and the formation of the UreI/urease membrane complex will provide novel eradication targets. 2. Identify the acid-dependent membrane proteome and the contribution of ArsS to intrabacterial trafficking of the identified proteins. This remarkable example of intra-bacterial protein trafficking provides a unique opportunity to develop anti-infective drugs. 3. Identify high affinity ArsS inhibitors by High- Throughput Screening (HTS). Because the acid-sensing domain of ArsS is in the periplasm and ArsS is essential for gastric infection, it is an attractive eradication target. We have designed an HTS that will allow detection of inhibitors that select between ArsS and UreI. The work of aims 1 and 2 will use biochemical and molecular biological techniques to identify proteins and regulatory pathways responsible for maintenance of gastric colonization by H. pylori. The work proposed in aim 3 will provide lead compounds that inactivate the ArsRS signaling cascade required for gastric infection. The results of these three aims will have an important positive impact by providing novel targets and improving current therapies for eradication.

 描述（由申请人提供）：幽门螺杆菌定植于世界上约 50% 人口的正常分泌酸的胃中。定植与胃病有关，包括胃炎、消化性溃疡和十二指肠溃疡、胃癌和 MALT 淋巴瘤。尽管三联疗法的初始根除率是成功的（~95%），但抗生素耐药性使得成功治疗感染变得越来越困难。因此，迫切需要一种不含抗生素的替代根除单一疗法。幽门螺杆菌特异性、保留共生肠道菌群或改进抗生素治疗，减少治疗的持续时间和复杂性。 这项研究的长期目标是了解幽门螺杆菌如何响应胃酸并在胃酸中存活，并利用这些知识来开发新的治疗方案和/或改进当前的根除疗法。这里的目的是确定酸敏感双组分系统 (TCS) ArsRS 如何调节酸诱导的脲酶及其辅助蛋白向内膜的运输，以与胃感染和胃酸存活所需的 UreI 形成膜结合复合物。该研究的基本原理是，了解 ArsS 使用的机制将允许药理干扰，从而产生新的幽门螺杆菌特异性无抗生素单一疗法和/或改进当前的根除方案。 具体目标是： 1. 阐明由传感器激酶ArsS介导的酸诱导信号传导机制。由于 ArsS 对于胃定植和胃酸存活至关重要，因此了解蛋白质转运至 UreI 的机制以及 UreI/脲酶膜复合物的形成将提供新的根除靶点。 2. 鉴定酸依赖性膜蛋白质组以及 ArsS 对所鉴定蛋白质的细菌内运输的贡献。这个细菌内蛋白质运输的显着例子为开发抗感染药物提供了独特的机会。 3.识别高 通过高通量筛选 (HTS) 筛选亲和力 ArsS 抑制剂。由于 ArsS 的酸敏感结构域位于周质中，并且 ArsS 对于胃感染至关重要，因此它是一个有吸引力的根除目标。我们设计了一种 HTS，可以检测在 ArsS 和 UreI 之间进行选择的抑制剂。 目标 1 和 2 的工作将使用生化和分子生物学技术来识别负责维持幽门螺杆菌胃定植的蛋白质和调节途径。目标 3 中提出的工作将提供使胃感染所需的 ArsRS 信号级联失活的先导化合物。这三个目标的结果将通过提供新的靶点和改进现有的根除疗法产生重要的积极影响。