CORE 2: Technology Core

核心2：技术核心

• 批准号: 10549999
• 负责人: Danielle L Swaney
• 金额: $ 40.2万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-17 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10549999
• 关键词: 2019-nCoV; 3-Dimensional; A549; Acetylation; Affinity Chromatography; Alveolar; Back; Bacterial Infections; Biochemical; Bioinformatics; Biological Assay; Biological Sciences; Cell Line; Cell Separation; Cells; Classification; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Communicable Diseases; Complex; Core Facility; Cryoelectron Microscopy; Crystallography; Data; Dedications; Deuterium; Disease; Disease Outcome; Education; Equipment; Generations; Genetic; Genetic Screening; Genomics; Goals; Human; Hydrogen; IL17 gene; Infection; Infrastructure; Integration Host Factors; Knock-out; Label; Laboratories; Macrophage; Maps; Mass Spectrum Analysis; Measures; Mentors; Microscopy; Modeling; Molecular; Mus; Mycobacterium tuberculosis; Organoids; Pathogenesis; Patients; Peripheral; Phosphorylation; Physiological; Post-Translational Protein Processing; Proteins; Proteomics; Research Personnel; Research Project Grants; Research Support; Resolution; Resources; Respiratory Disease; Sampling; Serum; Services; Severity of illness; Signal Transduction; Site; Small Interfering RNA; Structure; System; Systems Biology; Techniques; Technology; Training; Validation; Variant; Viral; Virus Diseases; Visualization; Work; X-Ray Crystallography; airway epithelium; beamline; clinically relevant; crosslink; data management; electron crystallography; genetic approach; genome-wide; innovation; insight; interest; knock-down; method development; model building; new technology; new therapeutic target; outreach; pathogen; protein complex; protein expression; protein protein interaction; protein purification; respiratory; respiratory pathogen; respiratory virus; structural biology; targeted treatment; therapeutic development; three dimensional cell culture; tissue culture; tool; variants of concern

TECHNOLOGY CORE SUMMARY Systematic characterization of protein and genetic interaction networks of respiratory pathogens in disease- relevant cells and 3D culture is critical for the identification of functional host-pathogen complexes and targets for therapeutic development. Systems biology approaches that determine protein-protein interactions (PPIs), characterize protein and posttranslational modification (PTM) abundance changes during infection, and classify functional host factors that regulate infectious disease pathogenesis, can provide strategic insight and generate models of respiratory pathogen infection. Combined with visualization of key host-pathogen complexes at atomic level through structural biology, and integrating with clinical and patient data related to disease severity, our innovative “systems to mechanism, bedside to benchtop” approach can be incredibly powerful for the identification of new therapeutic targets and for predicting respiratory disease outcome. The Technology Core will provide the infrastructure and technological expertise essential to the completion of the overall Host Pathogen Map Initiative (HPMI) 2.0 objectives. With support of Core facilities, the Technology Core will provide a suite of state-of-the-art proteomic technologies (Thermo Fisher Scientific Proteomics Facility for Disease Target Discovery, UCSF, Quantitative Biosciences Institute (QBI), Gladstone Institute), genetic screens (Innovative Genomics Institute, UCBerkeley), and structural biology approaches (UCSF Advanced Microscopy Laboratory and Crystallography Facilities, UCSF) for the generation of high-quality data, which will be integrated in collaboration with the Data Management and Bioinformatics and Modeling Core with clinical and patient derived data. Our goal is to provide PPI and global proteomic analyses, genome-wide and targeted knockdown and knockout genetic screens, and structural characterization of host-pathogen interactions for Mycobacterium tuberculosis (Mtb) (Project 1), and respiratory viruses: SARS-CoV-2, IAV, IBV, RSV, and HPIV (Project 2). We will analyze selected variants and clinical isolates for each pathogen in disease-relevant cell lines, primary cells isolated from healthy donors or mice, 3D-culture models including human airway epithelial organoids, and infected patient samples. Finally, we will continue to develop, optimize, and innovate new technological advances as needed to support Project 1 and 2 and HPMI 2.0 overall goals.

技术核心 总结 疾病中呼吸道病原体的蛋白质和遗传相互作用网络的系统表征- 相关细胞和三维培养对于鉴定功能性宿主-病原体复合体和靶标至关重要 用于治疗发展。确定蛋白质-蛋白质相互作用的系统生物学方法 （PPI），表征感染期间蛋白质和翻译后修饰（PTM）丰度变化，以及 对调节感染性疾病发病机制的功能性宿主因子进行分类，可以提供战略性见解， 生成呼吸道病原体感染的模型。结合关键宿主-病原体的可视化 通过结构生物学在原子水平上的复合物，并与临床和患者相关数据相结合， 我们创新的“从系统到机制，从床边到实验室”的方法， 对于识别新的治疗靶点和预测呼吸系统疾病 结果。技术核心将提供必要的基础设施和技术专长， 完成宿主病原体图谱倡议（HPMI）2.0的总体目标。在Core的支持下 技术核心将提供一套最先进的蛋白质组学技术（Thermo Fisher Scientific疾病靶点发现蛋白质组学设施，UCSF，定量生物科学 研究所（QBI），格莱斯顿研究所），遗传筛选（创新基因组学研究所，加州大学伯克利分校）， 和结构生物学方法（UCSF高级显微镜实验室和晶体学 设施，加州大学旧金山分校），以产生高质量的数据，这将是在与合作集成 数据管理和生物信息学和建模核心与临床和患者衍生数据。我们 目标是提供PPI和全球蛋白质组学分析，全基因组和靶向敲除， 分枝杆菌基因敲除筛选和宿主-病原体相互作用的结构表征 结核病（Mtb）（项目1）和呼吸道病毒：SARS-CoV-2、IAV、IBV、RSV和HPIV（项目2）。 我们将分析疾病相关细胞系中每种病原体的选定变体和临床分离株， 从健康供体或小鼠分离的原代细胞，包括人气道上皮细胞的3D培养模型 类器官和受感染的病人样本最后，我们将不断开发、优化、创新 支持项目1和2以及HPMI 2.0总体目标所需的技术进步。