Vascular adhesion protein-1 (VAP-1) as a therapeutic target in stroke

血管粘附蛋白-1 (VAP-1) 作为中风治疗靶点

基本信息

批准号：
8013600
负责人：
DALE Alan PELLIGRINO
金额：
$ 33.66万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-01-15 至 2014-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8013600
关键词：
Accounting Adhesions Adult Affect Animals Antibodies Appearance Behavior Blood Blood Vessels Brain Cell Adhesion Molecules Cells Cerebrum Characteristics Clinical Clinical Trials Coupled Cytoplasmic Granules Dichloromethylene Diphosphonate Encapsulated Encephalitis Endothelial Cells Endothelium Enzymes Estrogens Exhibits Failure Female Flow Cytometry Hematogenous Hour Immunoglobulins Immunohistochemistry Infarction Infiltration Inflammatory Inflammatory Response Integrins Intervention Ischemia Ischemic Stroke Laboratories Leukocytes Link Liposomes Lymphocyte Measurement Mediating Middle Cerebral Artery Occlusion Modeling Monitor Mononuclear Leukocytes Neurons Neutrophil Infiltration Pathway interactions Pattern Peripheral Pharmaceutical Preparations Play Population Process Prosencephalon Proteins Protocols documentation Publishing Rattus Recovery Reperfusion Therapy Reporting Rodent Rodent Model Role Selectins Stimulus Stroke Surface Surgical sutures T-Lymphocyte Testing Therapeutic Time Tissues base clinical efficacy design diabetic diamino oxhydrase drug efficacy improved inhibitor/antagonist intravital microscopy male monocyte neurobehavioral neuropathology neuroprotection neurotoxic neutrophil novel prevent public health relevance research study response therapeutic target trafficking vascular bed

项目摘要

DESCRIPTION (provided by applicant): Vascular adhesion protein-1 (VAP-1), also called semicarbazide-sensitive amine oxidase (SSAO), is reported to play an important role in the adhesion and endothelial transmigration of multiple leukocyte subsets (i.e., neutrophils, monocytes, lymphocytes) in peripheral vascular beds. Our laboratory recently reported that pharmacologic blockade of VAP-1/SSAO, during the reperfusion period following transient forebrain ischemia (TFI), prevented neutrophil infiltration into the brain, and was associated with a 6h post-ischemic therapeutic window in animals where heightened neutrophil diapedesis (which begins at ~6h reperfusion) had been documented. These results strongly suggested that post-TFI neutrophil infiltration and neuropathology were linked. However, the rats used in that study (diabetic, estrogen-treated ovariectomized females) are known to exhibit an exaggerated post-ischemic neutrophil infiltration response and, therefore, such findings may not necessarily be taken to suggest a beneficial role for VAP-1/SSAO inhibition over a wide spectrum of rodent models of ischemia/reperfusion. In the present project, we will examine the effect of a highly- selective inhibitor of VAP-1/SSAO, LJP-1586. Treatments will be initiated at 0-24h reperfusion in adult male rats subjected to 1h MCAo (intraluminal suture) and 1-14 days recovery. The proposal is guided by two Specific Aims designed to test the following hypotheses: (1) VAP-1/SSAO blockade will provide long-term neuroprotection (i.e., reduced infarct volumes; improved neurobehavioral function) with extended therapeutic windows that may, in part, be linked to diminished expression/activation of selected adhesion molecules. (2) The neuroprotection will largely involve interference with non- neutrophil leukocyte subsets. Experiments will involve selective depletion of neutrophils (anti-PMNL antibody) and monocytes (liposome-encapsulated clodronate), coupled with analysis of post-MCAo intracerebral expression of neutrophils, hematogenous monocytes, and T-lymphocytes using immunohistochemistry and FACS analyses. Preliminary evidence supports both hypotheses, suggesting a restricted role for neutrophils, but favoring a significant contribution from non-neutrophil subsets (i.e., mononuclear leukocytes) in the neuropathology accompanying temporary MCAo. Published evidence that mononuclear leukocytes do not display increased intracerebral adhesion and transmigration until many hours (even days) following the appearance of neutrophils may account for the long therapeutic windows (6-12h) we have observed in these early experiments (based upon results using 2 different, but selective, VAP- 1/SSAO blockers). The characterization of a validated pharmacologic approach that affects multiple leukocyte subsets, but with limited off-target actions, has substantial translational implications, since it may prove to be efficacious over a wide range of clinical stroke and ischemic presentations. PUBLIC HEALTH RELEVANCE: Post-ischemic brain inflammation can play a major role in the cerebral neuropathology that occurs following stroke. One potentially key player in that inflammatory attack is the leukocyte. These blood-derived cells may enter the brain after a stroke and release neurotoxic agents. The failure of anti-leukocyte strategies in clinical trials to date may, in part, relate to the presence of multiple leukocyte subsets and redundancy in the pathways responsible for their passage into the brain. The present project utilizes a selective pharmacologic approach that targets a novel protein which appears to be important in the trafficking of all leukocyte subsets and, therefore, may prove to have wide-ranging clinical efficacy.

描述（申请人提供）：血管粘附蛋白-1（VAP-1），也称为氨基脲敏感胺氧化酶（SSAO），据报道在多种白细胞亚群（即中性粒细胞）的粘附和内皮迁移中发挥重要作用、单核细胞、淋巴细胞）位于外周血管床中。我们的实验室最近报道，在短暂前脑缺血 (TFI) 后的再灌注期间，VAP-1/SSAO 的药物阻断可阻止中性粒细胞渗入大脑，并与动物的缺血后 6 小时治疗窗口相关，其中中性粒细胞渗出增加（在再灌注约 6 小时开始）已被记录。这些结果强烈表明 TFI 后中性粒细胞浸润与神经病理学之间存在关联。然而，该研究中使用的大鼠（糖尿病、雌激素治疗的卵巢切除雌性）已知表现出夸大的缺血后中性粒细胞浸润反应，因此，这些发现不一定表明 VAP-1/ 具有有益作用。 SSAO 对多种啮齿动物缺血/再灌注模型的抑制作用。在本项目中，我们将研究 VAP-1/SSAO 的高选择性抑制剂 LJP-1586 的效果。治疗将在成年雄性大鼠再灌注 0-24 小时时开始，进行 1 小时 MCAo（腔内缝合）并恢复 1-14 天。该提案以两个具体目标为指导，旨在测试以下假设：(1) VAP-1/SSAO 阻断将提供长期神经保护（即减少梗塞体积；改善神经行为功能），并延长治疗窗，部分可能，与所选粘附分子的表达/激活减少有关。 (2)神经保护主要涉及对非中性粒细胞亚群的干扰。实验将包括选择性去除中性粒细胞（抗 PMNL 抗体）和单核细胞（脂质体封装的氯膦酸盐），并使用免疫组织化学和 FACS 分析分析 MCAo 后脑内中性粒细胞、血源性单核细胞和 T 淋巴细胞的表达。初步证据支持这两种假设，表明中性粒细胞的作用有限，但支持非中性粒细胞亚群（即单核白细胞）在伴随暂时性 MCAo 的神经病理学中的显着贡献。已发表的证据表明，单核白细胞直到中性粒细胞出现后数小时（甚至数天）才表现出脑内粘附和迁移增加，这可能解释了我们在这些早期实验中观察到的长治疗窗口（6-12小时）（基于使用2的结果）不同但具有选择性的 VAP-1/SSAO 阻滞剂）。影响多种白细胞亚群但脱靶作用有限的经过验证的药理学方法的表征具有重大的转化意义，因为它可能被证明对广泛的临床中风和缺血性表现有效。公共卫生相关性：缺血后脑部炎症在中风后发生的脑神经病理学中发挥重要作用。炎症攻击中一个潜在的关键角色是白细胞。这些血液来源的细胞可能在中风后进入大脑并释放神经毒性物质。迄今为止，临床试验中抗白细胞策略的失败可能部分与多种白细胞亚群的存在以及负责其进入大脑的途径的冗余有关。本项目采用选择性药理学方法，针对一种新型蛋白质，该蛋白质似乎对所有白细胞亚群的运输都很重要，因此可能被证明具有广泛的临床功效。