Chemical biology of type IV secretion systems

IV型分泌系统的化学生物学

• 批准号: 10333392
• 负责人: Carrie Shaffer
• 金额: $ 29.91万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10333392
• 关键词: 2-hydroxypyridine; Address; Antibiotic Resistance; Bacteria; Bacterial DNA; Bacterial Genome; Basic Science; Biochemical; Biochemical Genetics; Biogenesis; Biological; Biological Assay; Biology; C10; Cells; Cellular biology; Centers of Research Excellence; Chemicals; Clinical; Collaborations; Complement; Complex; DNA; Destinations; Development; Discipline; Engineering; Epithelial Cells; Escherichia coli; Eukaryotic Cell; Exhibits; Faculty; Fostering; Foundations; Genetic; Goals; Helicobacter pylori; Impairment; Infection; Infectious Diseases Research; Knowledge; Lead; Libraries; Ligand Binding; Lipids; Lipopolysaccharide Biosynthesis Pathway; Mechanics; Mediating; Medical; Microscopy; Molecular; Molecular Machines; Molecular Probes; Nucleic Acids; Nucleoproteins; Oncoproteins; Organic Synthesis; Outcome; Pathogenesis; Pathogenicity Island; Peptidoglycan; Pharmaceutical Chemistry; Pharmacologic Substance; Phenotype; Pilum; Plant Model; Play; Polysaccharides; Prokaryotic Cells; Proteins; Regulation; Research; Research Design; Resolution; Resources; Rhizobium radiobacter; Risk; Role; Series; Signal Pathway; Stomach; Stomach Diseases; Structure-Activity Relationship; System; Systems Biology; Therapeutic; Toxin; Type IV Secretion System Pathway; Validation; Virulence; Work; antimicrobial; appendage; base; carcinogenicity; cell envelope; design; extracellular; genetic approach; high throughput screening; inhibitor; innovation; insight; interdisciplinary approach; invention; malignant stomach neoplasm; microbial; microbiome; model development; mutant; nanomachine; novel; pathogen; pathogenic bacteria; pharmacophore; protein complex; rational design; repository; response; scaffold; scale up; screening; small molecule; system architecture; tool; tumorigenic; virtual screening; 2-hydroxypyridine; Address; Antibiotic Resistance; Bacteria; Bacterial DNA; Bacterial Genome; Basic Science; Biochemical; Biochemical Genetics; Biogenesis; Biological; Biological Assay; Biology; C10; Cells; Cellular biology; Centers of Research Excellence; Chemicals; Clinical; Collaborations; Complement; Complex; DNA; Destinations; Development; Discipline; Engineering; Epithelial Cells; Escherichia coli; Eukaryotic Cell; Exhibits; Faculty; Fostering; Foundations; Genetic; Goals; Helicobacter pylori; Impairment; Infection; Infectious Diseases Research; Knowledge; Lead; Libraries; Ligand Binding; Lipids; Lipopolysaccharide Biosynthesis Pathway; Mechanics; Mediating; Medical; Microscopy; Molecular; Molecular Machines; Molecular Probes; Nucleic Acids; Nucleoproteins; Oncoproteins; Organic Synthesis; Outcome; Pathogenesis; Pathogenicity Island; Peptidoglycan; Pharmaceutical Chemistry; Pharmacologic Substance; Phenotype; Pilum; Plant Model; Play; Polysaccharides; Prokaryotic Cells; Proteins; Regulation; Research; Research Design; Resolution; Resources; Rhizobium radiobacter; Risk; Role; Series; Signal Pathway; Stomach; Stomach Diseases; Structure-Activity Relationship; System; Systems Biology; Therapeutic; Toxin; Type IV Secretion System Pathway; Validation; Virulence; Work; antimicrobial; appendage; base; carcinogenicity; cell envelope; design; extracellular; genetic approach; high throughput screening; inhibitor; innovation; insight; interdisciplinary approach; invention; malignant stomach neoplasm; microbial; microbiome; model development; mutant; nanomachine; novel; pathogen; pathogenic bacteria; pharmacophore; protein complex; rational design; repository; response; scaffold; scale up; screening; small molecule; system architecture; tool; tumorigenic; virtual screening

PROJECT SUMMARY Bacteria have evolved specialized nanomachines to deliver microbial cargo across the cell envelope. One versatile translocation apparatus, the type IV secretion system (T4SS), can be strategically deployed to inject macromolecular substrates into target bacterial or eukaryotic recipient cells. Despite their importance in bacterial pathogenesis and dissemination of antibiotic resistance determinants, the mechanisms by which the T4SS assembles and transports payload remain largely undefined. To address this knowledge gap, the long-term goal of this proposal is to develop and apply robust molecular tools to accelerate fundamental studies of T4SS nanomachines. The cag T4SS of the gastric bacterium Helicobacter pylori has emerged as an important system for understanding how a single molecular machine can transport diverse cargo into target cells. Whereas some T4SS have the capacity to secrete hundreds of proteins or DNA-protein complexes into the host cell, the ability to translocate a diverse repertoire of lipid, nucleic acid, protein, and polysaccharide substrates distinguishes the cag T4SS from other systems. Notably, the bacterial oncoprotein CagA is rapidly delivered to host gastric cells via cag T4SS mechanisms. Translocated H. pylori effector molecules activate innate defenses and dysregulate signaling pathways that influence progression of gastric disease; consequently, colonization by cag T4SS- positive H. pylori significantly augments the risk for gastric cancer. As a result of its central role in bacterial pathogenesis, the T4SS represents an ideal target for antimicrobials. In this application, we propose to identify and mechanistically characterize novel small molecule-based T4SS modulators. Iterative structure-activity relationship studies will be used to develop chemical scaffolds and pharmacophores with optimized anti-virulence potential. Probe development will take advantage of expertise and assay platforms in the Shaffer lab and will leverage synergistic resources in the proposed CPRI Computational Core (ligand-binding model development, rational design, virtual screening), CPRI Translational Core (high throughput assay support, novel compound repositories, and ADMET profiling), and the Organic Synthesis Core (medicinal chemistry and scale-up). Prioritized and validated chemical probes will be used in conjunction with biochemical and genetic approaches to interrogate cag T4SS regulation and dynamic steps in substrate translocation. Using a similar multidisciplinary approach, we will determine how the H. pylori cag T4SS apparatus assembles at the bacteria-host cell interface. Collectively, these studies will stimulate new basic research directions and will provide important insight into how the T4SS nanomachine orchestrates the delivery of specific molecular cargo to target cells to drive microbial pathogenesis. Furthermore, this work will generate powerful chemical tools that are broadly applicable to infectious disease research, and will identify potent lead compounds with the potential to disarm T4SS function in a variety of medically-relevant pathogens.

项目摘要 细菌已进化出专门的纳米机器，以在细胞包膜上传递微生物货物。一 多功能易位设备，IV型分泌系统（T4SS），可以策略性地部署 大分子底物进入靶标细菌或真核受体细胞。尽管它们在细菌中很重要 抗生素耐药性决定因素的发病机理和传播，T4SS的机制 组装和运输有效载荷仍然不确定。为了解决这个知识差距，长期目标 该建议的是开发和应用强大的分子工具来加速T4SS的基础研究 纳米机器。胃细菌幽门螺杆菌的CAG T4SS已成为重要的系统 为了了解单分子机器如何将各种货物运输到目标细胞中。而有些 T4S具有将数百种蛋白质或DNA蛋白复合物分泌到宿主细胞中的能力。 为了易位的脂质，核酸，蛋白质和多糖底物的多种曲目，将 来自其他系统的CAG T4S。值得注意的是，细菌癌蛋白CAGA迅速传递到宿主胃细胞 通过CAG T4SS机制。幽门螺杆菌效应子分子激活先天防御和失调 影响胃病进展的信号通路；因此，CAG T4SS-定植 幽门螺杆菌阳性显着增加了胃癌的风险。由于其在细菌中的核心作用 发病机理，T4SS代表了抗菌剂的理想靶标。在此应用程序中，我们建议确定 并机械地表征了新型的基于小分子的T4SS调节剂。迭代结构活动 关系研究将用于开发具有优化抗病毒性的化学支架和药算归因于 潜在的。探针开发将利用Shaffer Lab中的专业知识和测定平台，并将 在拟议的CPRI计算核心中利用协同资源（配体结合模型开发， 理性设计，虚拟筛选），CPRI翻译核心（高吞吐量测定支持，新颖的化合物 存储库和ADMET分析）以及有机合成核心（药物化学和扩大规模）。 优先和经过验证的化学探针将与生化和遗传方法一起使用 询问CAG T4SS调节和底物易位的动态步骤。使用类似的多学科 方法，我们将确定幽门螺杆菌CAG T4SS设备如何在细菌宿主宿主细胞界面组装。 总的来说，这些研究将刺激新的基础研究方向，并将提供重要的见解 T4SS纳米机械策划了特定的分子货物向靶细胞的递送以驱动微生物 发病。此外，这项工作将生成强大的化学工具，这些工具广泛适用于 传染病研究，并将确定有效的铅化合物，并具有解除T4SS功能的潜力 在各种与医学相关的病原体中。