Alternative Mechanisms of Monocyte Transendothelial Migration in Inflammation

炎症中单核细胞跨内皮迁移的替代机制

• 批准号: 9759026
• 负责人: Margarette Helen Clevenger
• 金额: $ 4.04万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9759026
• 关键词: Academia; Acute; Address; Affect; Air; Anti-inflammatory; Area; Atherosclerosis; Biological Assay; Blood Circulation; Blood Vessels; CD31 Antigens; Cell Adhesion Molecules; Cells; Chronic; Confocal Microscopy; Data; Disease; Endothelial Cells; Event; Flow Cytometry; Gene Expression Profile; Goals; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Kinesin; Knock-out; Lateral; Learning; Left; Leukocyte Adhesion Molecules; Leukocytes; Light; Lymphocyte; Mediating; Membrane; Membrane Protein Traffic; Methods; Microtubules; Migration Assay; Modeling; Molecular; Movement; Mus; Myocardial Infarction; Pathologic; Pathology; Pathway interactions; Phase; Phenotype; Population; Process; Property; Receptor Signaling; Recycling; Refractory; Reperfusion Injury; Research; Research Personnel; Residual state; Resistance; Role; Signal Pathway; Signal Transduction; Site; Stimulus; Stroke; Structure; Surface; Therapeutic; Time; Tissues; Ursidae Family; Vasculitis; body system; career; cell motility; cell type; clinical application; combat; comorbidity; experience; fighting; in vitro Model; in vivo; innovation; intravital microscopy; migration; monocyte; mouse model; neutrophil; pathogen; recruit; response; screening; tissue repair; trafficking; transcriptome sequencing; wound healing

Project Summary Inflammation serves a protective role in the body to fight infection and mediate wound healing; however, the inflammatory response can become deleterious if left uncontrolled or unresolved. Most pathology, including atherosclerosis, vasculitis, and ischemia/reperfusion injury—which can exacerbate potential comorbidities such as myocardial infarcts and stroke—is due to dysfunctional inflammation. Targeting inflammation could thus provide the means to combat inflammatory disease. This objective can be realized by targeting a common process that underlies all inflammation: the directed movement of white blood cells (WBCs) from circulation into affected tissue. In this process, transendothelial migration (TEM), WBCs migrate between lateral borders of endothelial cells (ECs) lining blood vessels to enter sites of inflammation. TEM is a critical step in the inflammatory response as it is the ultimate step preceding WBC access to inflamed tissue. Successful blocking of TEM would preclude tissue damage in an inflammatory context and is thus a therapeutic goal. TEM is mediated by interactions between surface adhesion molecules on the WBC and ECs, resulting in signaling within ECs that traffics membrane from the EC lateral border recycling compartment (LBRC), a perivascular vesiculotubular structure, to the site of TEM. This provides additional surface area required for WBC migration between adjacent ECs. Trafficking of LBRC is the final common pathway for TEM regardless of WBC type or inflammatory stimulus. Blocking critical adhesion molecules or downstream EC signaling pathways blocks TEM of WBC by 80-90% in vitro and in vivo. However, residual transmigration rate remains 10-20%, raising the question as to whether specific subpopulation(s) of WBC are resistant to TEM blockade and whether non-blocked WBCs use alternative signaling pathways to recruit the LBRC to transmigrate. AIM I will phenotypically characterize monocytes that overcome TEM blockade to determine if they represent a distinct monocyte subset. I will interrogate the mechanism by which these cells transmigrate in an in vitro TEM model and in murine models of inflammation. This will allow for the identification of surface receptors/signaling pathways that can be targeted to block the exit of all circulating monocytes from the vasculature in situations in which blocking 100% of inflammation is critical. AIM II will interrogate mechanisms that allow these monocytes to use alternative transmigration pathways. I will utilize a guided in vitro screening approach and in vivo real time intravital confocal microscopy to validate my in vitro results by inspecting mechanisms functioning at the TEM step directly. The project will shed light on alternative TEM pathways that may be important under some pathologic conditions and alert us to ways in which the body may compensate for the chronic blockade of traditional TEM pathways.

项目摘要 炎症在体内发挥了受保护的作用，以抵抗感染并介导伤口愈合。 但是，如果不受控制或未解决，炎症反应可能会变得有害。大多数病理学， 包括动脉粥样硬化，血管炎和缺血/再灌注损伤，这可能会加剧潜力 合并症，例如心肌实际上和中风，这是由于功能失调的炎症。定位 因此，炎症可以为打击炎症性疾病提供手段。这个目标可以通过 针对构成所有炎症的共同过程：白细胞的定向运动 （WBC）从循环到受影响的组织中。在此过程中，跨内皮迁移（TEM），WBC迁移 在内皮细胞（ECS）衬里血管的侧向边界之间进入炎症部位。 tem是一个 炎症反应的关键步骤是WBC进入发炎组织之前的最终步骤。 成功阻断TEM将在炎症环境中排除组织损害，因此是 治疗目标。 TEM是由WBC和EC上表面粘合分子之间的相互作用介导的，导致 在EC内的信号传导中，来自EC外侧边界回收室（LBRC）的交通膜，A 血管周围的囊泡结构，到TEM的部位。这提供了所需的其他表面积 WBC在相邻的EC之间迁移。 LBRC的贩运是TEM的最终公共途径 WBC类型或炎症刺激。阻止临界粘合分子或下游EC信号传导 途径在体外和体内将WBC的TEM限制为80-90％。但是，剩余的传输率仍然存在 10-20％，提出了一个问题，即WBC的特定亚群是否对TEM封锁有抵抗力 以及非阻滞的WBC是否使用替代信号通路来募集LBRC发射。 目的，我将表型典型地表征单核细胞，以克服TEM封锁以确定它们是否是否 代表一个独特的单核细胞子集。我将询问这些细胞在 体外TEM模型和炎症的鼠模型。这将允许识别表面 接收器/信号通路可以针对以阻止所有循环单核细胞的出口 在阻塞100％炎症的情况下的脉管系统至关重要。 AIM II将审问机制 这使这些单核细胞可以使用替代传输途径。我将利用引导的体外筛查 接近和体内实时插入式共聚焦显微镜，通过检查来验证我的体外结果 直接在TEM步骤工作的机制。该项目将阐明替代的TEM途径 在某些病理状况下可能很重要，并提醒我们身体可以补偿的方式 对于传统TEM途径的慢性封锁。