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

• 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
• 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

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)：血管黏附蛋白-1(VAP-1)，也被称为氨基脲敏感胺氧化酶(SSAO)，据报道在外周血管床上多种白细胞亚群(即中性粒细胞、单核细胞、淋巴细胞)的黏附和内皮迁移中发挥重要作用。我们实验室最近报道，在短暂性前脑缺血(TFI)后再灌注期，药物阻断VAP-1/SSAO可防止中性粒细胞 在动物的脑缺血后6h的治疗窗口中，中性粒细胞的高度脱失(开始于~6h的再灌流)已被记录在案。这些结果有力地表明，TFI后中性粒细胞浸润与神经病理有关。然而，这项研究中使用的大鼠(糖尿病、雌激素治疗的去卵巢雌性)已知表现出缺血后中性粒细胞渗透反应的夸大，因此，这些发现可能不一定表明在广泛的啮齿动物缺血/再灌注模型中抑制VAP-1/SSAO具有有益的作用。在本项目中，我们将研究高度选择性的VAP-1/SSAO抑制剂LJP-1586的效果。成年雄性大鼠在MCAO(腔内缝合)1h再灌注1~14d后开始治疗。该提案以两个具体目标为指导，旨在检验以下假设：(1)阻断VAP-1/SSAO将提供长期的神经保护(即，减少梗死体积；改善神经行为功能)，延长治疗窗口，这可能部分与所选黏附分子的表达/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。实验将包括选择性地去除中性粒细胞(抗PMNL抗体)和单核细胞(脂质体包裹的氯屈肼)，并利用免疫组织化学和FACS分析来分析MCAO后脑内中性粒细胞、造血单核细胞和T淋巴细胞的表达。初步证据支持这两种假说，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的神经病理中的重要贡献。已发表的证据表明，直到中性粒细胞出现后数小时(甚至数天)，单核细胞才显示出脑内黏附和移行增加，这可能是我们在这些早期实验中观察到的长时间治疗窗口(6-12h)的原因(基于使用两种不同的、但有选择性的VAP-1/SSAO阻滞剂的结果)。一种有效的药理学方法影响多个白细胞亚群，但具有有限的非靶点作用，其特征具有实质性的翻译意义，因为它可能被证明在广泛的临床中风和缺血表现中是有效的。 公共卫生相关性：脑缺血后炎症可在中风后发生的脑神经病理中发挥重要作用。在这种炎症攻击中，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，释放神经毒剂。到目前为止，临床试验中抗白细胞策略的失败可能在一定程度上与多个白细胞亚群的存在以及负责它们进入大脑的通路中的冗余有关。本项目利用一种选择性的药理学方法，针对一种新的蛋白质，该蛋白质似乎在所有白细胞亚群的运输中具有重要作用，因此可能被证明具有广泛的临床疗效。