Systems Approach to Immunity and Inflammation

说明(申请人提供)：免疫反应是一把双刃剑；它是宿主防御所必需的，但不受监管，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得到回答。这项提议试图揭示新的调控机制，以控制对NIAID优先病原体的先天和适应性反应的关键方面。这份U19报告提出了一种新的、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，同时消除了之前的几个障碍。这种协同作用的结果是基因发现和随后的机械性研究的范围和速度都有了显着的提高。这一计划是由下一代测序技术的快速进步和新的系统生物学方法实现的，这些技术通过允许在表型筛选之前识别ENU诱导的突变而改变了遗传学，并且通过新的系统生物学方法允许在基因调节和信号网络中对突变进行情景分析。这个集成的U19计划中的多个核心将支持和支持三个互补的项目，这些项目在几个垂直方向上探测免疫系统。UTSW小组将对多种PAMP进行无偏见的免疫反应筛查，这些PAMP是多种NIAID优先病原体的组成部分。该小组将利用每个家系中存在的突变知识来加速在这些筛查中识别出的基因。ANU小组将选择性地丰富将被系统生物学分析预测为在适应性免疫反应中发挥作用的基因突变的小鼠群体，并分析这些基因在感染NIAID优先病原体流感和外皮病病毒期间的作用。西雅图生物医学小组将全面筛选巨噬细胞和树突状细胞的体外缺陷，以应对广泛的天然免疫激动剂。该小组将重点关注通过外显子组测序确定存在ENU诱导基因突变的小鼠，系统分析预测这些基因是免疫细胞网络的关键调节因素，并将确定这些基因在对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.