Anesthetics and Stress Proteins in Brain Cell Injury

描述（由申请人提供）：卒中是美国第三大死亡原因，也是神经系统残疾的最常见原因。虽然许多工作都集中在对大脑神经元的损伤上，但从这个角度开发的疗法基本上都没有在临床试验中证明有效。最近的研究强调了神经胶质细胞在正常大脑和中风等损伤中发挥的重要和多方面的作用。长期目标是了解星形胶质细胞和小胶质细胞在中风中的有益和有害作用，以确定减少脑损伤的新靶点。70 kDa家族的热休克蛋白已被证明在中风的动物模型中具有保护作用，但迄今为止，该家族中只有一个成员，即胞质Hsp72，已被广泛研究。已知中风后的脑损伤涉及氧化应激和钙超载，这两者都与线粒体和内质网的损伤有关。这两种细胞器在决定脑细胞是否能在缺血性损伤中存活方面起着关键作用，因为它们密切参与处理缺血性损伤的标志性钙超载和自由基。本提案的第一个目的是研究两个Hsp70家族成员定位于这些细胞器的程度，Hsp75线粒体和Hsp78内质网，保护脑免受局灶性缺血和调节胶质细胞活化。这一目标包括通过改变线粒体代谢功能和氧化应激来调节炎症的研究。第二个目标是针对单个神经胶质细胞类型对中风的贡献。当在所有细胞类型中表达时具有保护性的胞质Hsp72将选择性地靶向星形胶质细胞或小胶质细胞，以确定仅限于这些细胞的过表达是否足以保护脑和保存神经元。小胶质细胞和星形胶质细胞的活化在对中风的炎症反应中是重要的，并且已经显示Hsp72过表达调节这种反应并减少损伤。将确定靶向表达对炎症的影响。由于在缺血期间细胞功能的许多方面同时改变，因此迄今为止，很难确定决定结果的关键途径和机制。在第三个目标中，将开发炎症和凋亡细胞死亡信号通路的动态计算模型，包括Hsp70的调制。这将允许使用计算工具来识别中风触发的分子信号相互作用中使用的关键控制策略。该模型将通过生物学实验进行验证，然后用于识别响应于炎症和缺血应激而诱发的调节策略。这种动态计算分析将确定干预最有可能强烈影响结果的点。通过从几个互补的角度来处理这个问题，将确定新的大脑保护候选人。中风是美国第三大死亡原因，也是神经系统残疾的最常见原因。这项研究的目的是确定减少中风引起的脑损伤的新方法，当转化为临床使用时，可以通过减少中风引起的损害来改善中风患者的生活。

DESCRIPTION (provided by applicant): Stroke is the third leading cause of death and the most common cause of neurological disability in the US. Although much work has focused on damage to neurons in the brain, essentially none of the therapies developed from this perspective has proven efficacious in clinical trials. Recent work has highlighted the important and multifaceted roles that glial cells play both in normal brain and following insults like stroke. The long term goal is to understand both the beneficial and deleterious effects of astrocytes and microglia in stroke to identify new targets to reduce brain injury. Heat shock proteins of the 70 kDa family have been shown to be protective in animal models of stroke, but to date only a single member of this family, the cytosolic Hsp72, has been extensively studied. Brain injury following stroke is known to involve oxidative stress and calcium overload, both of which are implicated in damage to both mitochondria and the endoplasmic reticulum. Both of these organelles play key roles in determining whether brain cells can survive ischemic injury, because they are intimately involved in dealing with the calcium overload and the free radicals that are hallmarks of ischemic injury. The first aim of this proposal will study the extent to which two Hsp70 family members localized to these organelles, Hsp75 to mitochondria and Hsp78 to endoplasmic reticulum, protect the brain from focal ischemia and modulate glial activation. This aim includes studies of modulation of inflammation by alteration of mitochondrial metabolic function and oxidative stress. The second aim targets the contribution of individual glial cell types to stroke. The cytosolic Hsp72, which is protective when expressed in all cell types, will be targeted selectively to either astrocytes or microglia to determine if overexpression restricted to these cells is sufficient to protect the brain and preserve neurons. Activation of microglia and astrocytes is important in the inflammatory response to stroke, and Hsp72 overexpression has been shown to modulate this response and reduce injury. The effect on inflammation of targeted expression will be determined. Because many aspects of cell function are altered simultaneously during ischemia, it has been difficult, to date, to identify the key pathways and mechanisms that determine outcome. In the third aim a dynamical computational model of inflammatory and apoptotic cell death signaling pathways, including modulation by Hsp70, will be developed. This will allow the use of computational tools to identify critical control strategies used in the molecular signaling interactions triggered by stroke. The model will be validated by biological experiments, and then used to identify the regulatory strategies evoked in response to inflammatory and ischemic stress. This dynamical computational analysis will identify the points at which intervention is most likely to strongly influence outcome. By approaching the problem from several complementary angles, new candidates for brain protection will be identified. PUBLIC HEALTH RELEVANCE Stroke is the third leading cause of death and the most common cause of neurological disability in the US. The goal of this research is to identify new ways to reduce the brain injury caused by stroke which when translated to clinical use could improve the lives of stroke victims by reducing the damage caused by stroke.