CORE 2: Technology Core

技术核心 摘要 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3D培养对于鉴定功能寄主-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)丰度的变化，以及 对调节传染病发病机制的功能性宿主因素进行分类，可以提供战略性的洞察和 建立呼吸道病原体感染模型。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形成复合体，并与临床和患者相关数据相结合 对于疾病的严重性，我们创新的“从系统到机制，从床边到台面”的方法可以 在识别新的治疗靶点和预测呼吸系统疾病方面极其强大 结果。技术核心将提供必要的基础设施和技术专业知识 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技术(Thermo 费舍尔疾病靶点发现科学蛋白质组学研究所，定量生物科学 基因筛查(加州大学伯克利分校创新基因组研究所)、 和结构生物学方法(加州大学旧金山分校高级显微镜实验室和结晶学 设施)，用于生成高质量的数据，这些数据将与 具有临床和患者衍生数据的数据管理、生物信息学和建模核心。我们的 目标是提供PPI和全球蛋白质组分析，全基因组和有针对性的敲除和 分枝杆菌的基因敲除筛选和宿主-病原体相互作用的结构特征 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个病原体的选定变种和临床分离株， 从健康供者或小鼠分离的原代细胞，包括人呼吸道上皮细胞的3D培养模型 有机物和受感染的病人样本。最后，我们将继续发展、优化、创新新 支持项目1和2以及HPMI 2.0总体目标所需的技术进步。

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.