Mechanisms Governing Monocyte Recruitment during Periodontal Inflammation

牙周炎症期间单核细胞募集的机制

• 批准号: 7792356
• 负责人: Rebecca Worthylake
• 金额: $ 10.54万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7792356
• 关键词: Actins; Adhesions; Area; Binding; Blood Circulation; CX3CL1 gene; Cell Adhesion Molecules; Cytoskeleton; Data; Development; Disease; Endothelium; GTP Phosphohydrolase Activators; Goals; Inflammation; Inflammatory; Inflammatory Response; Integrins; Kinetics; Leukocytes; Measures; Mediating; Modeling; Molecular; Oral health; Periodontal Diseases; Proteins; Proteomics; Reaction; Recruitment Activity; Regulation; Research Design; Research Personnel; Role; Signal Pathway; Signal Transduction; Testing; Therapeutic; Therapeutic Intervention; Tissues; Up-Regulation; base; chemokine; combat; design; in vivo; migration; monocyte; novel; public health relevance; research study; response; rho; rho GTP-Binding Proteins; trafficking

DESCRIPTION (provided by applicant): Inflammation is a major contributor to periodontal disease, and understanding the molecular mechanisms that govern the inflammatory response is a necessary precursor to the rational design of therapeutics to combat disease. Our overall objective is to discover specific signaling pathways that regulate monocyte recruitment to promote inflammation, and thereby contribute to the development and progression of periodontal disease. Chemokines are the molecular guides for monocyte trafficking throughout the body & the actin cytoskeleton and integrin adhesion are the molecular machinery that respond to the chemokine signals to effect monocyte migration. In our study, we are investigating how a chemokine associated with periodontal disease, CX3CL1, signals through the cytoskeletal regulator, ROCK, to promote monocyte recruitment from the circulation into periodontal tissue. In this proposal, we are pursuing two aspects of this signaling pathway. First, we are asking if CX3CL1-promoted adhesion is mediated by temporal regulation of ROCK activity. Our preliminary data shows that CX3CL1 and inhibition of ROCK both promote adhesion of monocytes to endothelial adhesion molecules. Chemokines are known to regulate ROCK and its upstream Rho GTPase activators, although the kinetics of specific Rho GTPases in response to CX3CL1 have not been investigated. The purpose of this aim will be to test the hypothesis that CX3CL1 signals through distinct Rho GTPases to transiently inhibit ROCK and promote monocyte adhesion to endothelial adhesion molecules. Second, we are investigating novel mechanisms of ROCK signaling by asking how ROCK interacts with a potential downstream effector, Coronin1A. We have identified Coronin1A as a putative ROCK interacting protein through a proteomics-based approach, and recent studies of Coronin1A function in leukocytes reveal an important role for Coronin1A in leukocyte adhesion, spreading and trafficking in vivo. The functional overlap of ROCK and Coronin1A, in combination with our proteomics results, strongly suggests a previously unrecognized functional interaction between these two proteins. The purpose of this aim will be to determine if ROCK binds to Coronin1A directly, or if Coronin1A is a substrate of ROCK. We anticipate that the results of Aim 1 will provide the data needed to convert our correlative relationship between CX3CL1, ROCK and adhesion into a direct mechanistic relationship. Similarly, while the case for interaction between ROCK and Coronin1A is compelling, the experiments in Aim 2 are necessary to provide the critical preliminary data to directly test their interaction. Together, the results of these aims will provide the necessary preliminary data needed to support a successful R01 application investigating the hypothesis that CX3CL1 triggers monocyte recruitment by inducing adhesion mediated by inhibition of ROCK and subsequent activation of Coronin1A. PUBLIC HEALTH RELEVANCE: Inflammation is a large contributor to periodontal disease, and developing strategies to combat pathological inflammatory reactions is a key goal for oral health investigators. Our studies are designed to understand the molecular mechanisms that govern periodontal inflammation, which will provide necessary information for rational development of therapeutic interventions.

描述（由申请人提供）：炎症是牙周病的主要原因，了解控制炎症反应的分子机制是合理设计治疗方法以对抗疾病的必要前提。我们的总体目标是发现调节单核细胞募集以促进炎症的特定信号通路，从而促进牙周病的发生和进展。趋化因子是单核细胞在体内运输的分子向导，肌动蛋白细胞骨架和整合素粘附是响应趋化因子信号以影响单核细胞迁移的分子机制。在我们的研究中，我们正在研究如何与牙周病相关的趋化因子，CX3CL1，信号通过细胞骨架调节，ROCK，促进单核细胞从循环到牙周组织的招聘。在这个提议中，我们正在研究这个信号通路的两个方面。首先，我们想知道CX3CL1促进的粘附是否是通过ROCK活性的时间调节介导的。我们的初步数据表明，CX3CL1和ROCK的抑制都促进单核细胞与内皮细胞粘附分子的粘附。已知趋化因子调节ROCK及其上游Rho GTP酶激活剂，尽管尚未研究响应于CX 3CL 1的特定Rho GTP酶的动力学。该目的的目的是检验CX3CL1通过不同的Rho GTP酶发出信号以瞬时抑制ROCK并促进单核细胞粘附于内皮粘附分子的假设。其次，我们正在研究ROCK信号转导的新机制，询问ROCK如何与潜在的下游效应子Coronin1A相互作用。我们已经确定Coronin1A作为一个假定的ROCK相互作用蛋白，通过蛋白质组学为基础的方法，和最近的研究Coronin1A在白细胞中的功能揭示了Coronin1A在白细胞粘附，扩散和运输在体内的重要作用。ROCK和Coronin1A的功能重叠，结合我们的蛋白质组学结果，强烈表明这两种蛋白质之间以前未被认识到的功能相互作用。该目的的目的是确定ROCK是否直接结合Coronin1A，或者Coronin1A是否是ROCK的底物。我们预期目标1的结果将提供将CX3CL1、ROCK和粘附之间的相关关系转换为直接机械关系所需的数据。同样，虽然ROCK和Coronin1A之间的相互作用是令人信服的，但目标2中的实验对于提供关键的初步数据以直接测试它们的相互作用是必要的。总之，这些目标的结果将提供必要的初步数据，以支持一个成功的R01应用调查的假设，CX3CL1触发单核细胞募集诱导粘附介导的抑制ROCK和随后的Coronin1A的激活。 公共卫生关系：炎症是牙周病的一个重要因素，制定策略来对抗病理性炎症反应是口腔健康研究人员的一个关键目标。我们的研究旨在了解牙周炎症的分子机制，这将为合理开发治疗干预措施提供必要的信息。