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.

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.我们的实验室最近报道说，在瞬时前脑缺血（TFI）之后的再灌注期间，VAP-1/SSAO的药理阻塞阻止了中性粒细胞 浸润到大脑中，并与动物的6小时缺血后治疗窗口有关，其中嗜中性粒细胞的尿症增强（从〜6h再灌注开始）已被记录。这些结果强烈表明，TFI后嗜中性粒细胞浸润和神经病理学是联系的。然而，已知该研究中使用的大鼠（糖尿病，雌激素治疗的卵巢切除女性）表现出夸张的夸张的缺血后的隔粒细胞浸润反应，因此，不一定要采取此类发现来提出对VAP-1/SSAO抑制rodent codent of Rodent/Reperfef的有益作用。在本项目中，我们将研究VAP-1/SSAO，LJP-1586的高度选择性抑制剂的效果。治疗将在接受1H MCAO（腔内缝合线）和1-14天恢复的成年雄性大鼠中以0-24H再灌注开始。该提案的指导是两个特定的目标，旨在检验以下假设：（1）VAP-1/SSAO阻滞将提供长期的神经保护（即减少梗塞量；改善神经行为功能），并通过扩展的治疗窗口与可能的延长的治疗窗口相关，可能会与所选择的表达/激活的表达性粘附呈现，以部分相关。 （2）神经保护在很大程度上涉及对非中性粒细胞白细胞亚群的干扰。实验将涉及嗜中性粒细胞（抗PMNL抗体）和单核细胞（脂质体包含的氯膦酸盐）的选择性消耗，并与中性粒细胞的脑内脑内表达分析，使用不受inmmphocytes分析中性粒细胞的脑内脑内表达。初步证据支持这两个假设，这表明中性粒细胞的作用受限制，但有利于在临时MCAO伴随神经病理学中的非中性噬细胞亚群（即单核白细胞）所产生的显着贡献。 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).影响多个白细胞亚群的经过验证的药理学方法的表征，但脱离靶向有限，具有实质性的转化含义，因为它可能被证明在广泛的临床中风和缺血性表现中是有效的。 公共卫生相关性：缺血后的大脑炎症可以在中风后发生的脑神经病理学中起主要作用。炎症性攻击的一个潜在关键人物是白细胞。这些血液衍生的细胞可能在中风后进入大脑并释放神经毒性剂。迄今为止，在临床试验中，抗白细胞策略的失败可能部分涉及多个白细胞子集的存在以及负责其进入大脑的途径中的冗余。本项目采用了一种选择性的药理学方法，该方法靶向一种新型蛋白质，该蛋白在所有白细胞亚群中似乎很重要，因此可能被证明具有广泛的临床疗效。