Mechanisms of CD40/CD40L in vascular injury and repair

CD40/CD40L在血管损伤与修复中的机制

• 批准号: 7910679
• 负责人: Guohong Li
• 金额: $ 32.96万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7910679
• 关键词: 3-nitrotyrosine; Address; Agreement; Angioplasty; Animal Model; Animals; Antibodies; Apolipoprotein E; Arterial Injury; Atherogenic Diet; Atherosclerosis; Attenuated; Biochemical; Blood Circulation; Blood Platelets; Blood Vessels; Bone Marrow; Bone Marrow Transplantation; CD40 Ligand; Cell Wall; Cells; Clinical; Consumption; Data; Development; Diabetes Mellitus; Environment; Genetic; Goals; Human; ITGAM gene; Immune; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Injury; Intravenous; Leukocyte Adhesion Molecules; Leukocyte-Adhesion Receptors; Leukocytes; Macrophage-1 Antigen; Mediating; Modeling; Mus; Neutrophil Activation; Pathway interactions; Patients; Plasma; Platelet Activation; Play; Probability; Process; Production; Proteins; Publishing; Recombinants; Relative (related person); Reporting; Role; Signal Transduction; Site; Stents; Superoxides; System; TNFRSF5 gene; Testing; Thromboplastin; Vascular Cell Adhesion Molecule-1; Vascular Diseases; Wild Type Mouse; Work; acute coronary syndrome; atherogenesis; base; coronary angioplasty; design; hypercholesterolemia; in vivo; injury and repair; insight; leukocyte activation; macrophage; mouse model; neointima formation; neutrophil; novel; novel therapeutics; prevent; receptor; reconstitution; research study; response; response to injury; restenosis

DESCRIPTION (provided by applicant): The long-term goal of this project is to better understand inflammatory mechanisms of vascular injury and repair and to develop new therapeutic strategies to treat patients with vascular diseases. Abundant evidence indicates that the CD40-CD40 ligand (CD40L) pathway plays a pivotal role in immune and inflammatory responses. Elevated levels of CD40L in the circulation have been associated with hypercholesterolemia, diabetes, acute coronary syndromes, and an increased probability of restenosis. To date there is no direct experimental evidence addressing if and how the CD40L pathway contributes to vascular injury and repair process. The results of our preliminary studies indicate multiple beneficial effects of CD40L blockade on the vascular injury response in animals on an atherogenic background, which is contrary to the limited evidence in a non-atherogenic mouse injury model. Based on our preliminary data, the overall hypothesis of this proposal is that circulating CD40L (both soluble CD40L and platelet-associated CD40L) is elevated and contributes to proinflammatory responses and neointimal formation after vascular injury in an atherogenic environment. To test our hypothesis and achieve our long-term goal, we will employ genetic, pharmacologic, and biochemical approaches in well-characterized cellular and animal models in the following three Specific Aims. Aim 1 will establish the role of CD40L in inflammation and neointima formation after arterial injury (carotid wire and FeCl3 injury) in apoE-/-CD40L-/- mice and apoE-/-CD40L+/+ mice vs in mice deficient in CD40L in a non-atherogenic environment. The relative contribution of CD40L expression by bone- marrow derived cells and vascular wall cells will be determined by creation of chimeric animals by bone marrow transplantation. Aim 2 will establish the contribution of soluble CD40L and platelet-associated CD40L in the inflammatory and thrombotic responses to vascular injury in an atherogenic milieu. We will examine platelet-leukocyte activation and interactions as well as tissue factor (TF) expression in apoE-/-CD40L-/- mice and apoE-/-CD40L+/+ mice, and then determine effects of intravenous reconstitution with recombinant sCD40L or CD40L-containing platelets in the mice on platelet-leukocyte leukocyte recruitment and TF expression at sites of arterial injury. Aim 3 will establish the role of CD40 (the receptor for CD40L) in neutrophil activation and inflammatory functions in vitro, and in the inflammatory and thrombotic responses to vascular injury in vivo. We will determine if CD40 is required for CD40L-mediated effects on neutrophils in vitro using CD40- expressing and CD40-deficient neutrophils, and then determine the role of CD40 in leukocyte recruitment and neointima formation after arterial injury in CD40-deficient (CD40-/- and apoE-/-CD40-/-) mice. In addition, we will determine if CD40 is required for CD40L-mediated TF induction in vivo. The results of these studies will provide mechanistic insight into the role of CD40/CD40L signaling in the vascular response to injury on an atherogenic background and may provide valuable information for the development of novel therapeutic strategies to prevent and treat vascular disease in humans. Project Narrative: Clinical restenosis, defined as e 50% loss of the initial lumen size of vessels, remains a major clinical problem in patients after coronary angioplasty and stent placement. This project is designed to better understand the mechanisms of restenosis using the well established mouse model of vascular injury and repair. The results obtained will provide novel insights into the mechanisms of restenosis and promise to provide valuable information for the development of novel therapeutic strategies to prevent and treat vascular disease in humans.

描述（由申请人提供）：该项目的长期目标是更好地了解血管损伤和修复的炎症机制，并制定新的治疗策略来治疗血管疾病患者。大量证据表明，CD40-CD40配体（CD40L）途径在免疫和炎症反应中起关键作用。循环中CD40L水平升高与高胆固醇血症，糖尿病，急性冠状动脉综合征以及再狭窄的可能性增加有关。迄今为止，尚无直接的实验证据表明CD40L途径是否以及如何促进血管损伤和修复过程。我们的初步研究的结果表明，CD40L阻断对动脉粥样硬化背景下动物血管损伤反应的多种有益作用，这与非动态小鼠损伤模型中的有限证据相反。根据我们的初步数据，该提案的总体假设是循环CD40L（可溶性CD40L和与血小板相关的CD40L）升高，并有助于促炎反应和在动脉典礼环境中血管损伤后的促炎反应和新内膜形成。为了检验我们的假设并实现我们的长期目标，我们将在以下三个特定目标中采用良好表征的细胞和动物模型中采用遗传，药理和生化方法。 AIM 1将在APOE-/ - / - CD40L - / - 小鼠和APOE-/ - /- CD40L+/+小鼠中，在非疗法环境中缺乏小鼠中，CD40L在APOE - / - CD40L - / - 小鼠和APOE-/ - / - CD40L - / - 小鼠和APOE - / - CD40L - / - 小鼠和APOE-/ - / - CD40L - / - 小鼠和APOE-/ - CD40L - / - 小鼠中，将确定CD40L在炎症和新内膜形成中的作用。骨髓衍生细胞和血管壁细胞表达CD40L表达的相对贡献将由骨髓移植创建嵌合动物。 AIM 2将建立可溶性CD40L和与血小板相关的CD40L在动脉粥样硬化环境中对血管损伤的炎症和血小板反应中的贡献。 We will examine platelet-leukocyte activation and interactions as well as tissue factor (TF) expression in apoE-/-CD40L-/- mice and apoE-/-CD40L+/+ mice, and then determine effects of intravenous reconstitution with recombinant sCD40L or CD40L-containing platelets in the mice on platelet-leukocyte leukocyte recruitment and TF expression at动脉损伤部位。 AIM 3将确定CD40（CD40L受体）在体外以及体内对血管损伤的炎症和血栓形成反应中的作用。 We will determine if CD40 is required for CD40L-mediated effects on neutrophils in vitro using CD40- expressing and CD40-deficient neutrophils, and then determine the role of CD40 in leukocyte recruitment and neointima formation after arterial injury in CD40-deficient (CD40-/- and apoE-/-CD40-/-) mice.此外，我们将确定CD40L介导的TF诱导体内是否需要CD40。这些研究的结果将提供有关CD40/CD40L信号在动脉粥样硬化背景上损伤的血管反应中的作用的机械洞察力，并可能为开发用于预防和治疗人类血管疾病的新型治疗策略提供宝贵的信息。项目叙述：临床再狭窄，定义为E血管初始管腔大小的50％损失，仍然是冠状动脉成形术和支架放置后患者的主要临床问题。该项目旨在通过良好的血管损伤和修复小鼠模型更好地了解再狭窄的机制。获得的结果将提供有关再狭窄机制的新见解，并有望为开发新的治疗策略提供有价值的信息，以预防和治疗人类的血管疾病。