Coordinated dynamic regulation and function of IRF transcription factors

项目摘要/摘要 巨噬细胞的天然免疫信号作为对病原体暴露的第一反应是至关重要的。三 干扰素调节因子(IRFs)是天然免疫反应的关键调节因子(即IRF3， ISGF3和IRF7)，它们对病原体暴露的反应是协调和相互依赖的。这些是如何 IRFS作为一种调节系统发挥作用，以及它们在调节广泛基因中的具体作用是什么 由数百个基因组成的表达程序仍然不清楚，因为以前的分析缺乏 在单细胞分辨率下定量IRF动力学的分辨率，以及这些因素之间的相互依赖 不能用流行的模型来描述。 因此，拟议研究的科学前提是利用尖端的量化单细胞和 全基因组的测量将允许我们开发一个定量的数学模型，描绘出 IRF3、ISGF3、IRF7各自的功能。我们将通过以下目标实现这一目标： 在目标1中，我们将阐明能够产生初级动态控制的复杂机制 我们在初步研究中揭示的响应因子IRF3。 在目标2中，我们将研究不同的、病原体特定的IRF3动力学对 染色质调控和靶基因表达。 在目标3中，我们将建立ISGF3的剂量响应和动态控制的预测信号模型 通过自分泌和旁分泌的I型干扰素的分泌激活，并表征其如何促进先天的 免疫基因在感染细胞和旁观者细胞中的表达。 在目标4中，我们将检查IRF7的监管控制，并检验它在 确定先前干扰素“警告”的细胞中的先天免疫反应，从而调节某种形式的先天免疫 免疫记忆。

Project Summary/Abstract Innate immune signaling by macrophages is critical as the first response to pathogen exposure. Three Interferon-regulatory factors (IRFs) are the critical regulators of the innate immune response (namely IRF3, ISGF3, and IRF7), and they respond to pathogen exposure coordinately and interdependently. How these IRFs function as a regulatory system, and what their specific roles are in regulating the extensive gene expression programs comprising hundreds of genes, has remained unclear, as previous assays lacked resolution to quantitate IRF dynamics at single cell resolution, and interdependencies between these factors could not be delineated by prevailing models. Thus the scientific premise of the proposed studies is that utilizing cutting-edge quantitative single cell and genome-wide measurements will allow us develop a quantitative mathematical model that delineates respective functions of IRF3, ISGF3, IRF7. We will address this goal with the following Aims: In Aim 1, we will elucidate the mechanism that are able to produce the complex dynamic control of the primary response factor IRF3 that we reveal in our preliminary studies. In Aim 2, we will investigate the functional consequence of differential, pathogen-specific IRF3 dynamics on chromatin control and target gene expression. In Aim 3, we will develop a predictive signaling model of the dose response and dynamic control of ISGF3 activated via autocrine and paracrine type I interferon secretion, and characterize how it contributes to innate immune gene expression in both infected and in bystander cells. In Aim 4, we will examine the regulatory control of IRF7 and test the hypothesis that it plays a key role in determining innate immune responses in previously interferon-`warned' cells, thus mediating a form of innate immune memory.