BIOLOGICAL EFFECTS OF DEATH REGULATORY GENES IN ISCHEMIA

死亡调节基因在缺血中的生物学效应

• 批准号: 6112625
• 负责人: ROGER Pancoast SIMON
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6112625
• 关键词: Alphaherpesvirinae; antisense nucleic acid; apoptosis; cerebral ischemia /hypoxia; gene induction /repression; neural degeneration; neurogenetics; neurons; tissue /cell culture; transcription factor; transfection /expression vector

Project 3: Biological effects of death-regulatory genes in ischemia In Project 3 we will directly test the hypothesis that the death of some neurons after ischemia results from the induced expression of specific death-promoter genes, while the survival of other neurons is promoted by the expression of death-suppressor genes. In Project 1, we have used the regional expression of known death-promoter and death-suppressor genes after ischemic injury to vulnerable or resistant brain regions to identify those genes which are likely to play a role in the survival or death of ischemic neurons; we have used subtractive hybridization to discover novel genes that may play a similar role. In Project 2 these neurons and novel candidate genes will be characterized in culture models of neuronal death. In order to directly test the role of these identified gene products in ischemic cell death, we will block expression of candidate death-promoter or death-suppressor genes in vivo using antisense oligonucleotides. Alternately, we will express candidate death-promoter or death-suppressor genes in vivo using a recombinant HSV-based vector. The effect of altered gene expression on cell survival following transient global ischemia and in ischemic tolerance (a model in which brief sub lethal focal ischemia protects neurons from subsequent prolonged ischemia) will be assessed histologically. The role of a putative death-promoter gene will be supported if blocking its expression increases cell survival after ischemia while overexpression increases the vulnerability of hippocampal neurons to ischemia. The role of a putative death-suppressor will be supported if blocking its expression blocks ischemic tolerance while overexpression reduces cell death following ischemia. The results of the studies in Project 3 will allow us to confirm the role of specific genes in ischemic cell death in vivo; this is a critical step in identifying candidate targets for novel therapies to reduce infarct size in stroke.

项目3：死亡调节基因在缺血中的生物学效应 在项目 3 中，我们将直接检验以下假设：某些人死亡 缺血后的神经元是由特定的诱导表达引起的 死亡促进基因，而其他神经元的存活则由 死亡抑制基因的表达。 在项目1中，我们使用了已知死亡启动子的区域表达 以及易受伤害或缺血性损伤后的死亡抑制基因 抗性大脑区域来识别那些可能发挥作用的基因 在缺血性神经元的存活或死亡中发挥作用；我们已经用过 消减杂交以发现可能发挥类似作用的新基因 角色。在项目 2 中，这些神经元和新的候选基因将被 神经元死亡培养模型的特征。 为了直接测试这些已鉴定的基因产物在 缺血性细胞死亡，我们将阻断候选死亡启动子的表达 或体内使用反义寡核苷酸的死亡抑制基因。 或者，我们将表达候选死亡促进者或死亡抑制者 使用基于 HSV 的重组载体进行体内基因检测。修改后的效果 短暂性局部缺血后基因表达对细胞存活的影响 缺血耐受（一种模型，其中短暂的亚致死局部缺血 保护神经元免受随后的长期缺血）将被评估 组织学上。假定的死亡促进基因的作用是 如果阻断其表达可增加细胞存活率，则支持 缺血而过度表达会增加海马的脆弱性 神经元缺血。假定的死亡抑制器的作用将是 如果阻断其表达会阻断缺血耐受性，则支持 过度表达可减少缺血后的细胞死亡。 项目 3 的研究结果将使我们能够确认该作用 体内缺血性细胞死亡中特定基因的研究；这是关键的一步 确定减少梗塞的新疗法的候选靶标 行程尺寸。