Regulation of Toll-like receptor-induced NFkB activation by Gai-coupled receptors

我的职业目标是领导一个独立的研究实验室，探索分子机制， 介导宿主防御和炎症。在炎症反应中，G蛋白偶联的 GPCR和先天性免疫受体同时被激活，并且每一种都可以激活多种不同的细胞。 蜂窝信号总体假设是炎症性GPCR信号传导与炎症性GPCR信号传导相互作用， 巨噬细胞Toll样受体（TLR）激活途径的许多成分，最终增强 促炎性转录因子核因子κ B（NF κ B）的活化。都支持这一看法 通过初步发现，白三烯B4（LTB 4）连接其GPCR BLT 1增强TLR-1， NFkB的依赖性激活。GPCR和TLR信号之间存在许多潜在的相互作用 可能共同决定NFkB的激活状态。为了发现这种相互作用，我们 提出一种跨学科的方法，将实验与系统生物学相结合， 能够通过实验进行测试的综合网络。本提案的总体计划是 在我最初两年的使用BLT 1-TLR的指导培训中获得知识和技能 作为一个模型的相互作用，这将被扩展到其他类别的GPCR在独立的 相位为了验证我们的假设，提出了以下具体目标：K99时期;目标1：确定 LTB 4/BLT 1信号传导（激酶，ROI生成）对TLR依赖性NF κ B活化的重要性， 目的2：检查LTB 4/BLT 1信号传导对TLR/IL-1 β受体表达的影响 目标3：开发涉及信号程序的计算模型（方程） 在Mo TLR应答期间由BLT 1诱导。R 00期：目标4：根据 K99时期，我们想进一步研究Gai蛋白偶联受体的重要性， BLT 1在TLR诱导的NF κ B活化中的作用，目的5：计算模型细胞信号网络 (GPCR和TLR），其调节NFkB活化以形成可检验的假设。这 该提案将为炎症中巨噬细胞活化的协调提供新的见解， 会让我成长为一名独立调查员

My career goal is to direct an independent research laboratory exploring the molecular mechanisms mediating host defense and inflammation. During inflammatory responses, both G protein-coupled (GPCRs) and innate immune receptors are activated simultaneously, and each can activate diverse cellular signals. The overall hypothesis is that inflammatory GPCR signaling interacts with and modifies many components of the macrophage Toll-like receptor (TLR) activation pathway, ultimately enhancing activation of the pro-inflammatory transcription factor nuclear factor kappa B (NFkB). This is supported by the preliminary finding that leukotriene B4 (LTB4) ligation of its GPCR BLT1 enhances the TLR- dependent activation of NFkB. There are many potential interactions between GPCR and TLR signaling that might combinatorially determine the state of NFkB activation. To discover such interactions, we propose an interdisciplinary approach combining experimentation with systems biology to model integrative networks that are capable of being tested experimentally.The general plan of this proposal is to acquire knowledge and skills during my initial two years of mentored training using BLT1-TLR interactions as a model, which will be extended to other classes of GPCRs during the independent phase. To test our hypothesis, the following Specific Aims are proposed: K99 period; Aim 1: Determine the importance of LTB4/BLT1 signaling (kinases, ROI generation) to TLR-dependent NFkB activation, Aim 2: Examine the effects of LTB4/BLT1 signaling on the expression of TLRs/IL-1beta receptors and their adaptors, Aim 3: Develop computational models (equations) involving the signaling programs induced by BLT1 during Mo TLR responses. R00 period:Aim 4: Based on the findings obtained in the K99 period, we want to further investigate the importance of Gai protein-coupled receptors other than BLT1 in TLR-induced NFkB activation, Aim 5: Computationally model cellular signaling networks (GPCR and TLRs) that regulate NFkB activation in order to formulate testable hypotheses. This proposal will provide new insights into the coordination of macrophage activation in inflammation, and will foster my development into an independent investigator.