Technology Core

项目概要、技术核心： 这种创新的综合系统生物学应用旨在描述复杂的宿主/病原体相互作用 发生在肺泡水平，导致严重肺炎治疗反应失败。整体 技术核心的目标是为所有项目和数据提供样本处理和数据生成支持 肺炎治疗成功临床反应 (SCRIPT) 系统生物学中心的其他核心。我们的 由于我们拥有专业知识，Technology Core 将为 SCRIPT 研究的成功做出独特的贡献 流式细胞术、细胞分选、各种下一代测序技术，包括基因 RNA 测序 用于基因组 DNA 甲基化的表达谱分析和全基因组亚硫酸氢盐测序，以及人源化小鼠 建模。技术核心将处理临床样本、分离特定细胞群并执行 项目 1 以及数据管理和生物信息学核心的转录和表观遗传分析， 生成人源化 MSTRG 小鼠并对这些小鼠的肺泡巨噬细胞进行转录分析 用于项目 2 和建模核心的小鼠。 目标 1：从非非特异性巨噬细胞/单核细胞和 T 细胞中分离并进行免疫表型分析。 支气管镜支气管肺泡灌洗（NBBAL）和肺炎患者外周血荧光检测 激活细胞分选（FACS）。 目标 2：对从 NBBAL 中分离的巨噬细胞和 T 细胞亚群进行转录和表观遗传分析 来自肺炎患者。 目标 3：验证在肺炎患者中观察到的病原体特异性基因表达特征 人源化 MSTRG 小鼠模型。

Project Summary, Technology Core: This innovative integrated systems biology application seeks to delineate the complex host/pathogen interactions occurring at the alveolar level that lead to unsuccessful response to therapy in severe pneumonia. The overall goal of the Technology Core is to provide sample processing and data generation support for all projects and other cores in the Successful Clinical Response In Pneumonia Therapy (SCRIPT) Systems Biology Center. Our Technology Core is uniquely poised to contribute to the success of the SCRIPT Study, as we possess expertise in flow cytometry, cell sorting, various Next Generation Sequencing techniques, including RNA-seq for gene expression profiling and genome-wide bisulfite sequencing for genomic DNA methylation, and humanized mouse modeling. The Technology Core will process clinical samples, isolate specific cellular populations and perform their transcriptional and epigenetic profiling for Project 1 and the Data Management and Bioinformatics Core, and generate humanized MISTRG mice and perform transcriptional profiling of alveolar macrophages from these mice for Project 2 and the Modeling Core. Aim 1: To isolate and immunophenotype subsets of macrophages/monocytes and T cells from non- bronchoscopic bronchoalveolar lavage (NBBAL) and peripheral blood patients with pneumonia via fluorescence- activated cell sorting (FACS). Aim 2: To perform transcriptional and epigenetic profiling of macrophage and T cell subsets isolated from NBBAL from patients with pneumonia. Aim 3: To validate pathogen-specific gene expression signatures observed in patients with pneumonia using a humanized MISTRG mouse model.