Targeting active adaptors to control endothelial damage

靶向活性接头来控制内皮损伤

基本信息

批准号：
7224951
负责人：
SHENG WEI
金额：
$ 27.05万
依托单位：
H. LEE MOFFITT CANCER CTR & RES INST
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-05-01 至 2009-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7224951
关键词：
1-Phosphatidylinositol 3-Kinase Activated Lymphocyte Acute Adaptor Signaling Protein Area Autoimmune Diseases Binding Biochemical Blood Vessels Bronchoalveolar Lavage Fluid Cell Death Cell Line Cell physiology Cells Cessation of life Chronic Cytolysis Cytoplasmic Granules Data Development Disease Disease Management Disruption Endopeptidases Endothelial Cells Enzymes Exocytosis Family Functional disorder Gene Delivery Granzyme Heart Diseases Human Immune In Vitro Inflammation Inflammatory Laboratories Large granular lymphocyte Lead Lesion Lung Lymphocyte Lytic Mediating Membrane Mitogen-Activated Protein Kinases Movement Natural Killer Cells Pathway interactions Patients Peptide Hydrolases Pharmacologic Substance Phosphotransferases Play Process Protease Inhibitor Proteins Reaction Receptor Activation Regulation Resistance Rheumatoid Arthritis Role Serine Protease Serine Proteinase Inhibitors Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Molecule Surface TYROBP gene Testing Therapeutic Intervention Tissues base cell injury chronic T-cell leukemia design drug development enzyme activity extracellular neoplastic cell novel therapeutics perforin primary pulmonary hypertension pulmonary artery endothelial cell therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Upon cell-cell contact, one of the mechanisms of immune cells to cause tissue damage is by delivery of lytic granules containing preformed effector molecules--perforin proteins and granzyme proteases. The presence of granzymes and perforin in the extracellular milieu not only reflects the presence of activated CTL and NK cells but also significantly contribute to inflammatory reaction. These protein enzymes can cause circular pore-like lesions on the membrane surface of endothelial cells and induce target cell death leading to local tissue damage and chronic vascular cell damage. Increased granzyme level is closely correlated with the inflammatory activity in autoimmune diseases (e.g. rheumatoid arthritis) and virally-infected lung and heart diseases. A clear understanding of the pathophysiology of inflammation mediated tissue damage would greatly facilitate management of this disease. The signal transduction pathways in effector lymphocytes, which trigger the redistribution of the lytic granules towards the target endothelial cells, are not well defined. Identification of key signaling molecules which specifically control this lytic process could enable pharmaceutical disruption of this process, thereby reducing the tissue damage mediated by activated lymphocytes. In this proposal, we will use a human primary pulmonary endothelial cell line, CRL- 2598, as the trigger to activate the lytic signal cascade in NK cells. Using biochemical and gene delivery approaches, we will directly test the hypothesis that the early signals via NK activation receptors and their associated adaptor proteins, DAP12 and DAP10, will play a specific role in control of granule movement in NK cells. Blocking of this initial step, at the level of the adaptor proteins, will inhibit granule exocytosis. More importantly, we will test our hypothesis on LGL leukemic patients with primary pulmonary hypertension (PPH) and determine if DAP10 and/or DAP12 critically control NK cell- mediated endothelial cell damage and death. A better understanding of the signaling pathways that control granule movement and exocytosis will offer new opportunities, e.g. design of DAP10 and DAP12 antagonists, for therapeutic intervention to specifically control lymphocyte-mediated tissue damage.

描述（由申请人提供）：在细胞 - 细胞接触时，免疫细胞造成组织损伤的机制之一是通过递送含有预制效应子分子的裂解颗粒 - 掺杂蛋白和颗粒蛋白酶。细胞外环境中粒酶和穿孔蛋白的存在不仅反映了活化的CTL和NK细胞的存在，而且还显着导致炎症反应。这些蛋白质酶可以在内皮细胞的膜表面引起圆形孔状病变，并诱导靶细胞死亡，导致局部组织损伤和慢性血管细胞损伤。颗粒水平升高与自身免疫性疾病（例如类风湿关节炎）和病毒感染的肺和心脏病的炎症活性密切相关。对炎症介导的组织损伤的病理生理学有清晰的了解将极大地促进这种疾病的治疗。效应淋巴细胞中的信号转导途径触发裂解颗粒向靶内皮细胞的重新分布，并未得到很好的定义。特异性控制这种裂解过程的关键信号分子的鉴定可以使该过程的药物破坏，从而减少活化淋巴细胞介导的组织损伤。在此提案中，我们将使用人类原代肺部内皮细胞系CRL-2598作为激活NK细胞中裂解信号级联的触发。使用生化和基因递送方法，我们将直接检验以下假设：NK激活受体及其相关衔接蛋白DAP12和DAP10的早期信号将在控制NK细胞的颗粒运动中发挥特定的作用。在衔接蛋白水平上阻止这一初始步骤将抑制颗粒胞吐作用。更重要的是，我们将对原发性肺动脉高压（PPH）LGL白血病患者的假设进行测试，并确定DAP10和/或DAP12是否严格控制NK细胞介导的内皮细胞损伤和死亡。更好地了解控制颗粒运动和胞吞作用的信号传导途径将提供新的机会，例如DAP10和DAP12拮抗剂的设计，用于治疗干预，以专门控制淋巴细胞介导的组织损伤。