Systems Approach to Immunity and Inflammation

描述（由申请人提供）：免疫反应是一把双刃剑；它绝对是宿主防御所必需的，但不受管制，会导致炎症性疾病。关于适当调整这些途径的机制的一些关键问题仍然没有答案。本研究旨在揭示控制NIAID优先病原体先天和适应性反应关键方面的新调控机制。本U19提出了一种全新的、完全集成的免疫学研究方法，我们称之为系统引导正向遗传学。在这里，我们结合了传统的正向遗传学和系统生物学方法，保留了每种策略的优势，同时消除了以前的几个障碍。这种协同作用的结果是基因发现的范围和速度以及随后的机制研究的显著增加。该项目得益于下一代测序技术的快速发展，该技术通过允许在表型筛选之前识别enu诱导的突变，改变了正向遗传学，并通过新的系统生物学方法允许在基因调控和信号网络中对突变进行背景化。这个综合U19项目中的多个核心将启用并支持三个互补的项目，这些项目将在几个正交的方向上探测免疫系统。UTSW小组将对多种PAMPs的免疫反应缺陷进行无偏筛选，这些PAMPs是多种NIAID优先病原体的成分。该小组将利用每个谱系中存在的突变知识来加速这些筛选中发现的基因的鉴定。澳大利亚国立大学的研究小组将选择性地丰富小鼠群体，以筛选系统生物学分析预测的在适应性免疫反应中发挥作用的基因突变，并分析这些基因在感染NIAID优先病原体流感病毒和电虫病毒时的作用。西雅图BioMed团队将全面筛选巨噬细胞和树突状细胞对广泛的先天免疫激动剂反应的体外缺陷。该小组将专注于通过外显子组测序鉴定出的具有ENU诱导突变的基因的小鼠，系统分析预测这些基因是免疫细胞网络的关键调节因子，并将确定这些基因在NIAID优先病原体沙门氏菌，李斯特菌和流感的免疫反应中的作用。

DESCRIPTION (provided by applicant): The immune response is a double-edged sword; it is absolutely required for host defense, but unregulated, causes inflammatory disease. A number of critical questions regarding the mechanisms that appropriately tailor these pathways remain unanswered. This proposal seeks to uncover novel regulatory mechanisms that control critical aspects of the innate and adaptive response to NIAID priority pathogens. This U19 proposes a novel and fully integrated approach to immunological research that we have termed Systems-Guided Forward Genetics. Here, we combine traditional forward genetics and systems biology methodologies, retaining the advantages of each strategy while eliminating several previous obstacles. The result of this synergy is a significant increase in both the scope and pace of gene discovery and of subsequent mechanistic studies. This program is enabled by the rapid advances in next-generation sequencing technology, which have transformed forward genetics by allowing the identification of ENU-induced mutations prior to phenotypic screening, and by new systems biology approaches that allow contextualization of the mutations within gene regulatory and signaling networks. The multiple Cores within this integrated U19 program will enable and support three complementary Projects that probe the immune system in several orthogonal directions. The UTSW group will conduct an unbiased screen for defective immune responses to a variety of PAMPs that are components of multiple NIAID Priority pathogens. This group will use knowledge of the mutations present in each pedigree to accelerate the identification of genes identified in these screens. The ANU group will selectively enrich the colony of mice to be screened for mutations in genes that have been predicted by systems biology analyses to play a role in adaptive immune responses, and analyze the role of these genes during infection with the NIAID Priority pathogens influenza and ectromelia viruses. The Seattle BioMed group will comprehensively screen macrophages and dendritic cells for in vitro defects in response to a wide-range of innate immune agonists. This group will focus on mice identified by exome sequencing to have ENU induced mutations in genes predicted by systems analysis to be critical regulators of immune ceil networks and will establish the role of these genes in the immune response to the NIAID Priority pathogens Salmonella, Listeria, and influenza.