Identification of TLR signaling network

TLR信号网络的识别

• 批准号: 6921353
• 负责人: SERGEI S MAKAROV
• 金额: $ 224.3万
• 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/6921353
• 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 样受体在先天性和适应性免疫反应的启动中发挥着关键作用。 TLR 家族的成员识别保守的微生物结构并激活信号通路，从而产生针对微生物感染的免疫反应。所有 TLR 都会激活共同途径来诱导一组核心的刻板反应，例如炎症。然而，单个 TLR 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组成部分知之甚少。鉴于 TLR 在宿主防御中的重要性，剖析这些途径是合理设计免疫调节剂和佐剂的关键。为了解决 TLR 信号网络的复杂性，必须采用能够系统检查信号转导的技术。 ATTAGENE Inc. 开发了一种可逆遗传方法，可以筛选包含数万个 cDNA 的表达文库，以鉴定信号传导中间体。我们使用这种方法来鉴定介导白介素 1 诱导的转录因子 NF-kB 激活途径的许多新成分。我们的研究表明，可逆遗传方法为信号转导的系统性、全基因组识别提供了高度通用的工具。 这种综合方法不依赖于先入为主的观念，并且具有消除误报背景的内置程序。在本研究中，我们将采用可逆遗传方法来系统鉴定 TLR 信号网络的组成部分。我们的目标是 (1) 确定将 TLR 家族各个成员与转录因子 NF-kB 激活联系起来的信号转导组件； (2) 将这些成分注释为正/负和微分/共同中间体； (3) 在体外检查所鉴定的介质在先天免疫反应中的生物学功能； (4) 创建敲除动物模型，以评估已确定的介质作为调节对不同病原体（包括单核细胞增多性李斯特菌和鼠伤寒沙门氏菌）的先天性和适应性免疫反应的潜在靶标。该计划的成功实施应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性免疫调节剂和佐剂的开发。