Identification of TLR signaling network

TLR信号网络的识别

• 批准号: 6818795
• 负责人: SERGEI S MAKAROV
• 金额: $ 222万
• 依托单位: ATTAGENE, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-15 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6818795
• 关键词: Listeria; Listeria infections; RNA interference; Salmonella infections; Salmonella typhimurium; biological signal transduction; cell line; complementary DNA; cooperative study; gene expression; gene targeting; genetic library; genetic regulation; genetic screening; genetically modified animals; immune response; immunogenetics; immunomodulators; interleukin 1; laboratory mouse; macrophage; microorganism immunology; nuclear factor kappa beta; toll like receptor

DESCRIPTION (provided by applicant): Toll-like receptors play a critical role in the initiation of the innate and adaptive immune responses. Members of the TLR family recognize conserved microbial structures and activate signaling pathways that result in immune responses against microbial infections. All TLRs activate common pathways to induce a core set of stereotyped responses, such as inflammation. However, individual TLRs can also induce immune responses that are tailored to a given microbial infection. The mechanisms and components of these varied responses are poorly understood. Given the importance of TLRs in host defense, dissection of these pathways is key to the rational design of immunomodulators and adjuvants. To address the complexity of the TLR signaling network, it is imperative to put in place technologies enabling systematic examination of the signal transduction. ATTAGENE Inc. has developed a reversible genetic approach that affords screening expression libraries of tens of thousands of cDNAs to identify signaling intermediates. We used this methodology to identify a number of novel components of the pathways that mediate interleukin-1-inducible activation of the transcription factor NF-kB. Our studies indicate that the reversible genetic approach offers a highly versatile tool for a systematic, genome-wide identification of signal transduction. This comprehensive approach does not rely on preconceived notions and it has built-in procedures that eliminate false-positive background. In this study, we will adapt the reversible genetic approach to systematic identification of components of the TLR signaling network. Our objectives are (1) to identify the components of signal transduction that link individual members of the TLR family with activation of the transcription factor NF-kB; (2) to annotate those components as positive/negative and differential/common intermediates; (3) to examine biological functions of the identified mediators in innate immune responses in vitro; and (4) to create knock-out animal models in order to assess the identified mediators as potential targets to modulate the innate and adaptive immune responses to different pathogens, including Listeria monocytogenes and Salmonella typhimurium. Successful implementation of the proposed plan should provide comprehensive knowledge of molecular mechanisms controlling immune responses to pathogens, thus greatly facilitating the development of highly specific immunomodulators and adjuvants.

描述(由申请人提供)：Toll样受体在启动先天和获得性免疫反应中发挥关键作用。TLR家族的成员识别保守的微生物结构，并激活信号通路，从而对微生物感染产生免疫反应。所有的TLR都激活了共同的通路，以诱导一系列核心的刻板印象的反应，如炎症。然而，单个TLRs也可以诱导针对特定微生物感染的免疫反应。人们对这些不同反应的机制和成分知之甚少。鉴于TLRs在宿主防御中的重要性，对这些途径的剖析是合理设计免疫调节剂和佐剂的关键。为了解决TLR信令网络的复杂性，必须采用能够对信号转导进行系统检查的技术。Attagene Inc.已经开发出一种可逆的遗传方法，该方法可以筛选数万个cDNA的表达文库，以识别信号中间产物。我们使用这种方法来确定一些新的成分的途径，以中介白介素1诱导的转录因子核因子-kB的激活。我们的研究表明，可逆遗传方法为系统的、全基因组范围的信号转导鉴定提供了一个高度通用的工具。这种全面的方法不依赖于先入为主的概念，它具有消除假阳性背景的内置程序。在这项研究中，我们将采用可逆遗传方法来系统地识别TLR信令网络的组件。我们的目标是(1)确定将TLR家族的单个成员与转录因子NF-kB激活联系起来的信号转导组件；(2)将这些组件注释为阳性/阴性和差异/共同中间体；(3)在体外检测已识别的介体在天然免疫反应中的生物学功能；以及(4)建立敲除动物模型，以评估已识别的介体作为潜在靶标来调节对不同病原体的固有和获得性免疫反应，包括李斯特菌和鼠伤寒沙门氏菌。该计划的成功实施将提供控制病原体免疫反应的分子机制的全面知识，从而极大地促进高度特异的免疫调节剂和佐剂的开发。