Transglutaminase 2 decreases ischemic insult by attenuating hypoxic signaling.

转谷氨酰胺酶 2 通过减弱缺氧信号来减少缺血性损伤。

基本信息

批准号：
7609278
负责人：
Anthony J Filiano
金额：
$ 1.78万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-03-30 至 2009-05-16
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7609278
关键词：
ARNT protein Adenoviruses Affect Alteplase Animals Apoptosis Apoptotic Architecture Asses Attenuated BCL2/Adenovirus E1B 19kd Interacting Protein 3-Like Binding Biological Assay Biological Models Blood - brain barrier anatomy C57BL/6 Mouse Calcium Cause of Death Cell Death Cell Line Cell Nucleus Cells Cessation of life Condition Cultured Cells Data Development Effectiveness Environment Enzymes Event Genes Genetic Genetic Transcription Glucose Goals Green Fluorescent Proteins Guanosine Triphosphate Phosphohydrolases HIF1alpha protein Hour Human Hypoxia Hypoxia Inducible Factor Immunoblot Analysis Immunohistochemistry In Situ Infarction Infection Control Injury Ischemia Knockout Mice LacZ Genes Lactate Dehydrogenase Lactate Dehydrogenases Lead Life Ligation Luciferases Magnetic Resonance Imaging Measures Mediating Messenger RNA Modeling Mus Nerve Degeneration Neuraxis Neurites Neuroblastoma Neurons Oxygen Pathway interactions Pattern Permeability Play Point Mutation Polymerase Chain Reaction Predisposition Property Protein Overexpression Proteins Quality of life Rattus Reporter Response Elements Role Scaffolding Protein Signal Transduction Stroke Stroke Volume Techniques Testing Therapeutic Thrombolytic Therapy Time Transgenic Organisms United States Food and Drug Administration Up-Regulation Vascular Endothelial Growth Factors Weight attenuation base caspase-3 deprivation disability hypoxia inducible factor 1 improved middle cerebral artery mouse model mutant neuron loss novel post stroke protein expression response scaffold size small molecule success transcription factor transglutaminase 2 vector

项目摘要

DESCRIPTION (provided by applicant): Stroke is the third leading cause of death and a major cause of long term disability in the US. Inadequate success of current thrombolytic therapy demonstrates a critical need for new targets to improve quality of life post-stroke. Neurons adapt to decreased oxygen delivery subsequent to stroke by up-regulating genes controlled by the hypoxia inducible factor (HIF). This transcription factor is comprised of two subunits: HIF1 alpha and HIF1 beta. Recent studies show that increased HIF1 during ischemia results in upregulation of apoptotic genes. Our preliminary data indicates that Transglutaminase 2 (TG2) binds to HIF1 beta resulting in attenuation of HIF activation and decreased cell death during ischemia. Also, TG2 inhibits the upregulation of the HIF controlled proapoptotic gene Bnip3, but not VEGF. Our hypothesis is that TG2 will facilitate neuronal protection following oxygen and glucose deprivation (OGD) by attenuating apoptotic genes induced by HIF1. We propose to delineate the effects of TG2 on HIF signaling and ischemic cell death with 3 aims. 1) To test the hypothesis that TG2 is upregulated during OGD and decreases HIF-dependent pro-apoptotic signaling. We will use a neuroblastoma cell line (SH-SY5Y) that overexpresses wild type TG2 and two TG2 constructs with point mutations that differentially affect 2 unique functions (transamidating and GTPase) of the enzyme. Using a HIF signaling reporter assay and analysis of mRNA and protein expression patterns of HIF dependent genes, we will distinguish which properties of TG2 are important for regulating HIF signaling. 2) To test the hypothesis that TG2 protects neuronal cells from ischemic insult and which functions of TG2 are important. Using SH-SY5Y cells and primary cortical neurons we will measure OGD-induced cell death and upstream apoptotic events. 3) To test the hypothesis that TG2 leads to overall protection in a mouse model of stroke. We will use a permanent Middle Cerebral Artery ligation model in C57BL/6 mice that express endogenous TG2, overexpress TG2, or lack TG2. We will measure infarct volumes using T2 weighted MRI as well as identifying genes regulated by TG2 with appropriate techniques. Our preliminary data show that overexpression of TG2 decreases stroke infarct volumes by 33%. We hypothesize that TG2 will lead to overall protection against stroke.

描述（由申请人提供）：中风是美国的第三大死亡原因，也是美国长期残疾的主要原因。当前的溶栓疗法的成功不足表明，对新目标的迫切需要在势头后改善生活质量。神经元适应了通过由缺氧诱导因子（HIF）控制的上调基因（HIF）的上调基因而降低的氧气递送。该转录因子由两个亚基组成：HIF1 Alpha和HIF1 Beta。最近的研究表明，缺血期间的HIF1增加导致凋亡基因上调。我们的初步数据表明，转谷氨酰胺酶2（TG2）与HIF1β结合，导致HIF激活衰减并减少缺血期间细胞死亡。同样，TG2抑制了HIF控制的促凋亡基因BNIP3的上调，但不能抑制VEGF。我们的假设是，通过减弱HIF1诱导的凋亡基因，TG2将促进氧和葡萄糖剥夺（OGD）后的神经元保护。我们建议描述TG2对3个目标的HIF信号传导和缺血细胞死亡的影响。 1）测试TG2在OGD期间上调并降低HIF依赖性促凋亡信号传导的假设。我们将使用神经母细胞瘤细胞系（SH-SY5Y），该细胞系过表达野生型TG2和两个TG2构建体，其点突变会差异影响酶的2个独特功能（跨增强和GTPase）。使用HIF信号记者测定和HIF依赖基因的mRNA和蛋白质表达模式的分析，我们将区分TG2的哪些特性对于调节HIF信号传导很重要。 2）测试TG2保护神经元细胞免受缺血性损伤以及TG2的功能很重要的假设。使用SH-SY5Y细胞和原代皮质神经元，我们将测量OGD诱导的细胞死亡和上游凋亡事件。 3）检验了TG2导致小鼠中风模型中总体保护的假设。我们将使用表达内源性TG2，过表达TG2或缺乏TG2的C57BL/6小鼠中的永久性大脑连接模型。我们将使用T2加权MRI测量梗塞体积，并使用适当的技术鉴定由TG2调节的基因。我们的初步数据表明，TG2的过表达可将中风梗塞量减少33％。我们假设TG2将导致对中风的总体保护。