Technology Core

项目摘要、技术核心： 这一创新的集成系统生物学应用试图描绘复杂的宿主/病原体相互作用 发生在肺泡水平，导致对严重肺炎的治疗无效。整体而言 技术核心的目标是为所有项目提供样本处理和数据生成支持 肺炎治疗(SCRIPT)系统生物学中心成功临床反应的其他核心。我们的 技术核心公司独一无二地准备为剧本研究的成功做出贡献，因为我们拥有专业知识 在流式细胞术、细胞分选、各种下一代测序技术中，包括用于基因的RNA-seq 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 模特儿。技术核心将处理临床样本，分离特定的细胞群体，并执行 他们对项目1的转录和表观遗传学描述以及数据管理和生物信息学核心， 并产生人源化的MISTRG小鼠，并从这些小鼠中进行肺泡巨噬细胞的转录图谱 项目2和建模核心的MICE。 目的1：分离非霍奇金淋巴瘤患者外周血中巨噬细胞/单核细胞和T细胞亚群并进行免疫表型鉴定。 经纤维支气管镜支气管肺泡灌洗(NBBAL)和外周血荧光检测肺炎 激活细胞分选(FACS)。 目的2：对从NBBAL分离的巨噬细胞和T细胞亚群进行转录和表观遗传学分析 来自肺炎患者。 目的3：验证肺炎患者中观察到的病原体特异性基因表达特征。 人性化的MISTRG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.