Matrix metalloproteinase-9 mediated cell death in stroke

基质金属蛋白酶 9 介导的中风细胞死亡

• 批准号: 7577473
• 负责人: Eng H. Lo
• 金额: $ 33.88万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7577473
• 关键词: Amplifiers; Anoikis; Area; Astrocytes; Binding; Biological Models; Blood - brain barrier anatomy; Caspase; Cell Adhesion Molecules; Cell Culture Techniques; Cell Death; Cell Survival; Cells; Cerebral Ischemia; Cerebrum; Cessation of life; Chemicals; Clinical Data; Coagulation Process; Collagen; Complement; Complementary DNA; Data; Edema; Endothelial Cells; Endothelium; Extracellular Signal Regulated Kinases; Extravasation; Fibronectins; Gelatinase B; Glucose; Homeostasis; Hypoxia; In Vitro; Infarction; Inflammatory; Injury; Integrins; Intercellular adhesion molecule 1; Ischemia; Knock-out; Knockout Mice; Knowledge; LacZ Genes; Laminin; Lead; Ligation; Link; Luciferases; MAPK14 gene; MAPK8 gene; MEKs; Maps; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Mechanics; Mediating; Mediator of activation protein; Middle Cerebral Artery Infarction; Middle Cerebral Artery Occlusion; Mitogen-Activated Protein Kinases; Modeling; Molecular; Mus; National Institute of Neurological Disorders and Stroke; Neurons; Outcome; Oxygen; Pathology; Pathway interactions; Peptide Hydrolases; Pharmaceutical Preparations; Principal Investigator; Program Review Group; Proteolysis; RNA Interference; Regulation; Reperfusion Therapy; Reporter; Research Priority; Resistance; Role; SB 203580; SP600125; STAT1 gene; Signal Pathway; Signal Transduction; Stroke; System; Testing; Therapeutic; Tissue Inhibitor of Metalloproteinase-1; Transcription Factor AP-1; Transgenic Mice; Tumor Necrosis Factor-alpha; U-0126; Up-Regulation; cytokine; deprivation; extracellular; human TNF protein; improved; in vivo; inhibitor/antagonist; insight; integrin-linked kinase; kinase inhibitor; mouse model; mutant; neurovascular unit; overexpression; prevent; programs; promoter; protective effect; ras-GRF1; receptor; research study; response

DESCRIPTION (provided by applicant): The NINDS Stroke Program Review Group identified proteolytic pathology in the neurovascular unit as a high priority research area. In this project, we will investigate the mechanisms of matrix metalloproteinase-9 (MMP-9) mediated cell death in cells of the neurovascular unit (cerebral endothelial cells, astrocytes, neurons). We hypothesize that after ischemia, MMP-9 degrades neurovascular matrix and disrupts cell-matrix interactions, thus triggering anoikis-like cell death in endothelium, astrocytes, and neurons. Hence, targeting signaling pathways that mediate MMP-9 dysregulation or initiating a rescue of homeostatic matrix signaling may be viable therapeutic approaches for stroke. To test this hypothesis, we will pursue 3 Specific Aims. In Aim 1, we examine mechanisms of MMP-9 regulation in cell cultures of cerebral endothelial cells, astrocytes, and neurons. Responses to hypoxia/reoxygenation and oxygen-glucose deprivation will be assessed. The roles of three major MAP kinases (ERK, p38, JNK) as signaling mediators will be examined. Endothelial activation via adhesion molecules will be explored as a molecular amplifier. In Aim 2, we test the idea that MMP-9 upregulation disrupts matrix integrin signals and triggers anoikis like death in endothelial cells, astrocytes, and neurons. We will examine the roles of integrin linked kinase (ILK) and Akt cell survival signaling, and try to prevent caspase-mediated cell death by integrin rescue. Pharmacologic inhibitors, RNA interference, and mutant cells lacking MMP-9 or overexpressing the endogenous inhibitor TIMP1 will be used. In Aim 3, we test the hypothesis that targeting MAP kinase pathways in mouse models of focal cerebral ischemia are protective by down regulating ischemic MMP-9 responses. U0126, SB203580, and SP600125 will be used to inhibit ERK, p38 and JNK MAP kinase pathways respectively. To complement these drugs, we will also use molecular approaches involving a JIP1 cDNA to block JNK and a Ras-GRF knockout against ERK. To assess a specific role for MMP-9, experiments will be performed in MMP-9 knockouts, transgenic mice that overexpress the endogenous MMP-9 inhibitor TIMP1, and transgenic mice that express LacZ driven by the MMP-9 promoter. Our lab and others have established MMP-9 as a key extracellular protease that modifies neurovascular homeostasis. These proposed studies should help reveal the mechanisms of MMP-9 mediated cell death in stroke.

描述（由申请人提供）：NINDS中风计划审查小组将神经血管单元中的蛋白水解病理确定为高优先研究领域。在该项目中，我们将研究神经血管单元（脑内皮细胞，星形胶质细胞，神经元）细胞中基质金属蛋白酶9（MMP-9）介导的细胞死亡的机制。 我们假设缺血后，MMP-9降解神经血管基质并破坏细胞矩阵相互作用，从而触发内皮细胞，星形胶质细胞和神经元中的厌氧菌样细胞死亡。因此，靶向介导MMP-9失调或启动稳态基质信号的营救的信号通路可能是中风的可行治疗方法。为了检验这一假设，我们将追求3个具体目标。 在AIM 1中，我们检查了MMP-9调节脑内皮细胞，星形胶质细胞和神经元细胞培养的机制。将评估对缺氧/二氧化量和氧气剥夺的反应。将检查三个主要的MAP激酶（ERK，P38，JNK）作为信号介质的作用。通过粘附分子的内皮激活将作为分子放大器探索。在AIM 2中，我们测试了MMP-9上调会破坏基质整联蛋白信号和触发诸如内皮细胞，星形胶质细胞和神经元的死亡的观念。我们将检查整联蛋白连接激酶（ILK）和AKT细胞存活信号的作用，并试图防止通过整合素recRecue进行caspase介导的细胞死亡。将使用缺乏MMP-9或过表达内源性抑制剂TIMP1的药理学抑制剂，RNA干扰和突变细胞。在AIM 3中，我们检验了以下假设：靶向局灶性脑缺血小鼠模型中的MAP激酶途径通过降低调节缺血性MMP-9反应来保护。 U0126，SB203580和SP600125将分别用于抑制ERK，p38和JNK MAP激酶途径。为了补充这些药物，我们还将使用涉及JIP1 cDNA的分子方法阻止JNK和对ERK的RAS-GRF敲除。为了评估MMP-9的特定作用，将在MMP-9敲除，过表达内源性MMP-9抑制剂TIMP1的转基因小鼠中进行实验，以及由MMP-9启动子驱动的LACZ的转基因小鼠。 我们的实验室和其他实验室已经建立了MMP-9作为修饰神经血管稳态的关键细胞外蛋白酶。这些提出的研究应有助于揭示MMP-9中风中介导的细胞死亡的机制。

项目成果

Role of ERK map kinase and CRM1 in IL-1beta-stimulated release of HMGB1 from cortical astrocytes.

• DOI: 10.1002/glia.20982
• 发表时间: 2010-06
• 期刊: GLIA
• 影响因子: 6.2
• 作者: Hayakawa, Kazuhide;Arai, Ken;Lo, Eng H.
• 通讯作者: Lo, Eng H.

Plasma and brain matrix metalloproteinase-9 after acute focal cerebral ischemia in rats.

• DOI: 10.1161/strokeaha.109.554824
• 发表时间: 2009-08
• 期刊: Stroke
• 影响因子: 8.3
• 作者: Park KP;Rosell A;Foerch C;Xing C;Kim WJ;Lee S;Opdenakker G;Furie KL;Lo EH
• 通讯作者: Lo EH

Neuroprotective roles and mechanisms of neuroglobin.

• DOI: 10.1179/174313209x389866
• 发表时间: 2009-03
• 期刊: Neurological research
• 影响因子: 1.9
• 作者: Yu Z;Fan X;Lo EH;Wang X
• 通讯作者: Wang X

Effects of neuroglobin overexpression on mitochondrial function and oxidative stress following hypoxia/reoxygenation in cultured neurons.

• DOI: 10.1002/jnr.21826
• 发表时间: 2009-01
• 期刊: JOURNAL OF NEUROSCIENCE RESEARCH
• 影响因子: 4.2
• 作者: Liu, Jianxiang;Yu, Zhanyang;Guo, Shuzhen;Lee, Sun-Ryung;Xing, Changhong;Zhang, Chenggang;Gao, Yan;Nicholls, David G.;Lo, Eng H.;Wang, Xiaoying
• 通讯作者: Wang, Xiaoying

Induction of vascular endothelial growth factor and matrix metalloproteinase-9 via CD47 signaling in neurovascular cells.

• DOI: 10.1007/s11064-010-0159-6
• 发表时间: 2010-07
• 期刊: NEUROCHEMICAL RESEARCH
• 影响因子: 4.4
• 作者: Xing, Changhong;Arai, Ken;Park, Kyung-Pil;Lo, Eng H.
• 通讯作者: Lo, Eng H.