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 调节宿主防御和炎症。在炎症反应中，两种G蛋白偶联 (GPCRs)和先天免疫受体是同时激活的，每个受体都可以激活不同的 手机信号。总体假设是炎症性gpr信号相互作用并改变。 巨噬细胞Toll样受体(TLR)激活途径的许多成分，最终增强 激活促炎转录因子核因子kappaB(NFkB)。这是受支持的 根据初步发现，白三烯B4(LTB4)与其GPCRBLT1的连接可增强TLR- 依赖于NFkB的激活。GPCR和TLR信号之间存在许多潜在的相互作用 这可能联合决定NFkB的激活状态。为了发现这种相互作用，我们 提出了一种实验与系统生物学相结合的跨学科建模方法 能够进行实验测试的综合网络。这项提议的总体计划是 在我使用BLT1-TLR进行的最初两年的指导培训中获得知识和技能 相互作用作为一个模型，将在独立期间扩展到其他类别的GPCR 相位。为了验证我们的假设，提出了以下具体目标：K99阶段；目标1：确定 LTB4/BLT1信号转导(激酶、ROI生成)对TLR依赖的NFkB激活的重要性 目的：研究LTB4/BLT1信号通路对TLRs/IL-1β受体表达的影响。 它们的适配器，目标3：开发涉及信令程序的计算模型(方程式) 在Mo-TLR反应过程中由BLT1诱导。R00期间：目标4：根据在 K99期间，我们想要进一步研究GAI蛋白偶联受体的重要性 BLT1在TLR诱导的NFkB激活中，目标5：计算模型细胞信号网络 (GPCR和TLRs)，调节NFkB的激活，以制定可检验的假说。这 该提案将为炎症中巨噬细胞激活的协调提供新的见解，以及 会把我培养成一名独立的调查员。

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.