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: 8600326
• 负责人: DALE Alan PELLIGRINO
• 金额: $ 33.32万
• 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 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8600326
• 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.

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），据报道在外周血管床中多种白细胞亚群（即中性粒细胞、单核细胞、淋巴细胞）的粘附和内皮迁移中发挥重要作用。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/SSAO 抑制对广泛的缺血/再灌注啮齿动物模型具有有益作用。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），并延长治疗窗口，这可能部分与减少所选粘附分子的表达/激活有关。 (2)神经保护主要涉及对非中性粒细胞亚群的干扰。实验将涉及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）后才表现出脑内粘附和迁移增加，这可能是我们在这些早期实验中观察到的长治疗窗（6-12小时）的原因（基于使用2种不同但选择性的VAP-1/SSAO阻滞剂的结果）。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。

项目成果

Protective role of fingolimod (FTY720) in rats subjected to subarachnoid hemorrhage.

• DOI: 10.1186/s12974-015-0234-7
• 发表时间: 2015-01-27
• 期刊: Journal of neuroinflammation
• 影响因子: 9.3
• 作者: Xu HL;Pelligrino DA;Paisansathan C;Testai FD
• 通讯作者: Testai FD

VAP-1 blockade prevents subarachnoid hemorrhage-associated cerebrovascular dilating dysfunction via repression of a neutrophil recruitment-related mechanism.

VAP-1阻断通过抑制中性粒细胞募集相关机制来预防蛛网膜下腔出血相关的脑血管扩张功能障碍。

• DOI: 10.1016/j.brainres.2015.01.047
• 发表时间: 2015
• 期刊: Brain research
• 影响因子: 2.9
• 作者: Xu,Haoliang;Testai,FernandoD;Valyi-Nagy,Tibor;NPavuluri,Mani;Zhai,Fengguo;Nanegrungsunk,Danop;Paisansathan,Chanannait;Pelligrino,DaleA
• 通讯作者: Pelligrino,DaleA