Inducible Renitence in Macrophages

巨噬细胞中的诱导性记忆

• 批准号: 8852635
• 负责人: JOEL A SWANSON
• 金额: $ 29.47万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8852635
• 关键词: Address; Asbestos; Atherosclerosis; Autophagocytosis; Bacteria; Bacterial Infections; Calcium; Cell physiology; Cells; Cholesterol; Chronic; Crohn' s disease; Cytoplasm; Disease; Event; Experimental Models; Exposure to; Genes; Goals; Health; Human; Immunocompromised Host; Individual; Infection; Inflammation; Inflammatory; Interferons; Investigation; Kinetics; Left; Lipopolysaccharides; Listeria; Listeria monocytogenes; Lysosomes; Macrophage Activation; Macrophage Cell Biology; Measures; Mechanics; Membrane; Methods; Microbe; Molecular; Mus; Mycoses; Natural Immunity; Nitrogen; Parasitic infection; Particulate; Patients; Phagocytes; Phagolysosome; Process; Property; Proteins; Quantitative Evaluations; Regulation; Research; Resistance; Resistance to infection; Role; Silicon Dioxide; Stimulus; System; Testing; Therapeutic; Tissues; Tumor Necrosis Factor-alpha; Urea; Vacuole; Virulence; Virus Diseases; Work; Wound Healing; acid sphingomyelinase; antimicrobial; bacterial resistance; base; cytokine; design; in vitro Model; inhibitor/antagonist; lysosome membrane; macrophage; microbicide; nanoparticulate; novel; pathogen; pressure; reactive oxygen intermediate; repaired; resistance mechanism; seal; synaptotagmin VII; therapy design

DESCRIPTION (provided by applicant): Macrophages are essential to innate immunity to infections. Activation of macrophages by lipopolysaccharide (LPS) and cytokines such as interferon-? (IFN?) and tumor necrosis factor-� (TNF�) increases their microbicidal activities but also increases damage to tissues due to inflammation. As therapies which target chronic inflammation leave patients vulnerable to infections, new strategies are needed that can selectively increase macrophage antimicrobial activities. The long-term goal of this research is to devise such strategies through investigations of fundamental macrophage cell biology. This lab discovered recently that exposure of murine macrophages to bacteria, LPS, IFN? or TNF� leads to stabilization of their lysosomes against mechanical damage, a phenomenon termed "inducible renitence" or IR. As vacuolar membrane damage is essential to the virulence of many pathogenic microbes, to infection by viruses and to inflammation by micro-articulates, this novel phenomenon could potentially be exploited therapeutically. The objective of the present work is to define the cellular and molecular basis of inducible renitence. The central hypothesis is that renitence is induced by classical activation and consists of enhanced mechanisms of membrane damage-repair. The experimental model for these studies is a system in which macrophage lysosomes or phagolysosomes are subjected to controlled levels of physical perturbation, which allows quantitative evaluation of mechanisms that resist or repair damage. The central hypothesis will be tested by addressing three specific aims. The first aim will determine the conditions and factors which induce renitence in human and murine macrophages. Renitence will be measured in classically activated macrophages, alternatively activated (wound-healing) macrophages and regulatory macrophages, as well as macrophages treated with other agents. The second aim will determine the role of membrane damage-repair mechanisms in renitence. The kinetics of phagolysosome damage and repair will be measured and the contributions of vacuolar calcium, synaptotagmin VII and acid sphingomyelinase to renitence will be analyzed. The third aim will determine the role of renitence in macrophage resistance to infection by the Gram-positive intracellular pathogen Listeria monocytogenes, which normally scapes into cytoplasm by damaging vacuolar membranes. By defining the cellular and molecular basis of IR, this research will introduce a new strategy for manipulation of macrophage function. Therapies which increase renitence selectively could reduce inflammation due to micro-particulates or benefit immunosuppressed patients and individuals with chronic inflammatory diseases, such as therosclerosis and Crohn's disease.

描述(申请人提供)：巨噬细胞对感染的先天免疫力是必不可少的。内毒素和干扰素等细胞因子激活巨噬细胞。(干扰素？)而肿瘤坏死因子-�(肿瘤坏死因子-�)在增强其杀菌活性的同时，也增加了炎症对组织的损伤。由于针对慢性炎症的治疗方法使患者容易受到感染，因此需要有选择地增加巨噬细胞抗微生物活性的新策略。这项研究的长期目标是通过对基础巨噬细胞生物学的研究来设计这样的策略。本实验室最近发现，小鼠巨噬细胞暴露于细菌、内毒素、干扰素？或肿瘤坏死因子�导致其溶酶体对机械损伤的稳定，这种现象被称为“诱导性突变”或IR。由于液泡膜损伤对许多病原微生物的毒力、病毒感染和微关节炎症是必不可少的，这一新现象可能被开发用于治疗。本工作的目的是定义可诱导重复性的细胞和分子基础。中心假说是，重复性是由经典的激活引起的，并由膜损伤修复的增强机制组成。这些研究的实验模型是一个巨噬细胞溶酶体或吞噬溶酶体受到可控水平的物理扰动的系统，这允许对抵抗或修复损伤的机制进行定量评估。核心假设将通过解决三个具体目标来检验。第一个目标将确定在人和小鼠巨噬细胞中诱导突变的条件和因素。重复性将在经典激活的巨噬细胞、交替激活的(伤口愈合)巨噬细胞和调节性巨噬细胞以及用其他药物治疗的巨噬细胞中进行测量。第二个目标将确定膜损伤修复机制在重复性中的作用。将测量吞噬溶酶体损伤和修复的动力学，并将分析空泡钙、突触素VII和酸性鞘磷脂酶在重复性中的作用。第三个目标将确定复原性在巨噬细胞抵抗革兰氏阳性细胞内病原体单核细胞增生性李斯特菌感染中的作用，这种细菌通常通过破坏空泡膜进入细胞质。通过定义IR的细胞和分子基础，本研究将为巨噬细胞功能的调控提供一种新的策略。选择性地增加复发率的疗法可以减少因微粒引起的炎症，或使免疫抑制的患者和患有慢性炎症性疾病的患者受益，如动脉粥样硬化和克罗恩病。