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绘复杂的宿主/病原体相互作用 发生在肺泡水平，导致对重症肺炎治疗的不成功反应。整体 技术核心的目标是为所有项目提供样品处理和数据生成支持， 其他核心在肺炎治疗的成功临床反应（CNOT）系统生物学中心。我们 技术核心是独一无二的准备，以促进成功的研究，因为我们拥有的专业知识， 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 建模该技术核心将处理临床样本，分离特定的细胞群， 他们的转录和表观遗传分析项目1和数据管理和生物信息学核心， 并产生人源化MISTRG小鼠，并从这些小鼠中进行肺泡巨噬细胞的转录谱分析， 项目2和建模核心的小鼠。 目的1：分离和免疫表型的巨噬细胞/单核细胞和T细胞亚群， 支气管镜支气管肺泡灌洗（NBBAL）和外周血通过荧光检测肺炎患者- 活化细胞分选（FACS）。 目的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 从肺炎患者身上。 目的3：使用一种新的方法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.