Heat shock proteins in brain ischemia and stroke

脑缺血和中风中的热休克蛋白

• 批准号: 8044553
• 负责人: Midori A Yenari
• 金额: --
• 依托单位: VETERANS AFFAIRS MED CTR SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2014-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8044553
• 关键词: ATP phosphohydrolase; ATPase Domain; Acute; Astrocytes; Binding; Biological Preservation; Blood - brain barrier anatomy; Brain; Brain Edema; Brain Injuries; Brain Ischemia; Brain hemorrhage; Catastrophic Illness; Cell Culture Techniques; Cell Death Process; Cessation of life; Data; Edema; Endopeptidases; Endothelial Cells; Extracellular Matrix; Family; Gelatinase A; Genetic Transcription; Heat Stress Disorders; Heat shock proteins; Heat-Shock Proteins 70; Hemorrhage; Human; In Vitro; Injury; Interruption; Ischemia; Ischemic Brain Injury; Ischemic Stroke; Lead; Matrix Metalloproteinases; Mediating; Medical; Molecular Chaperones; Nature; Neurologic; Pathology; Peptide Hydrolases; Pharmaceutical Preparations; Plasmids; Population; Prevention; Process; Property; Proteins; Research; Risk; Source; Stimulus; Stroke; Stroke prevention; System; Translation Process; Translations; Veterans; Work; cell type; effective therapy; in vitro Model; in vivo; in vivo Model; mutant; overexpression; prevent; protective effect; protein aggregation; protein folding

DESCRIPTION (provided by applicant): Stroke is a common affliction among veterans, and treatments are few. Brain ischemia frequently causes disruption of the blood brain barrier (BBB) leading to brain edema and hemorrhage which can complicate an already catastrophic illness. Recent work in the field has implicated matrix metalloproteinases (MMPs), a family of Zn-dependent endopeptidases, in the breakdown of the extracellular matrix and BBB leading to brain edema and hemorrhage. Work by our labs and those of our collaborators' have focused on the protective potential of heat shock proteins, namely, the highly inducible 70 kD heat shock protein (HSP70). HSP70 appears to have cytoprotective properties by nature of its chaperone functions, presumably leading to enhancement of nascent protein folding and prevention of protein aggregation. However, work in related fields has shown that HSPs may also be involved in the expression and processing of MMPs. We recently showed that overexpression of HSP70 or its induction by heat stress reduced expression of MMPs in cultured astrocytes at the transcriptional and translational level. In addition to preventing MMP transcription and translation, we have preliminarily found that HSP70 can also prevent processing of pro-MMPs to their active form, suggesting an additional action of protection. In this application, we propose to further explore these observations that HSP70 protects the brain against stroke by preventing BBB disruption, edema and hemorrhage by inhibiting the expression, translation and processing of MMPs. Aim 1 will use cell cultures of brain derived endothelial cells and astrocytes to establish which cell types generate HSP70 and MMPs following various stimuli, and if HSP70 can prevent disruption of the BBB using in vitro models. Aim 2 will determine how HSP70 interacts with MMPs. HSP70 mutants lacking either functional ATPase or substrate binding domains will be studied. Aim 3 will explore the significance of HSPs on BBB integrity, brain edema/hemorrhage and MMPs will be studied using in vivo models of experimental stroke. PUBLIC HEALTH RELEVANCE: Stroke is a common affliction among veterans, and treatments are few. Stroke is frequently complicated by the disruption of the blood brain barrier (BBB) causing brain swelling and hemorrhage. In this project, we propose to study the potential beneficial effect of a protein that is increased in the brain after stroke, HSP70. HSP70 is not normally present in the brain except under stressful conditions such as stroke. However, further increasing HSP70 during stroke can reduce the amount of ensuing brain injury and prevent complications of stroke as well. We propose to explore how HSP70 prevents brain swelling and hemorrhage, in order that pharmacological strategies to increase HSP70 or to administer HSP70 itself might be a useful treatment for stroke in humans some day. Since stroke is a common medical problem among veterans, work from this proposal could potentially lead to treatments benefiting this population.

描述（由申请人提供）： 中风是退伍军人中常见的苦难，而治疗很少。脑缺血经常导致血脑屏障（BBB）的破坏导致脑水肿和出血，这可能会使已经灾难性疾病复杂化。该领域的最新工作暗示了基质金属蛋白酶（MMP），这是一个Zn依赖性内肽酶家族，在细胞外基质和BBB的分解中，导致脑水肿和出血。我们的实验室和合作者的工作集中在热休克蛋白的保护潜力上，即高度诱导的70 kD热休克蛋白（HSP70）。 HSP70的伴侣功能本质上具有细胞保护特性，可能导致了新生蛋白质折叠的增强并预防蛋白质聚集。但是，在相关领域的工作表明，HSP也可能参与MMP的表达和处理。我们最近表明，HSP70的过表达或通过热应力降低了转录和翻译水平培养的星形胶质细胞中MMP表达的诱导。除了防止MMP转录和翻译外，我们还先发现HSP70还可以防止将Pro-MMP处理为其活性形式，从而提出了另外的保护作用。在此应用中，我们建议进一步探索这些观察结果，即HSP70通过抑制MMP的表达，翻译和处理来防止BBB破坏，水肿和出血来保护大脑免受中风的影响。 AIM 1将使用脑衍生的内皮细胞和星形胶质细胞的细胞培养物来确定哪种细胞类型在各种刺激下产生HSP70和MMP，如果HSP70可以防止使用体外模型阻止BBB破坏BBB。 AIM 2将确定HSP70如何与MMP相互作用。将研究缺乏功能性ATPase或底物结合结构域的HSP70突变体。 AIM 3将使用实验中风的体内模型来研究HSP对BBB完整性，脑水肿/出血和MMP的重要性。 公共卫生相关性： 中风是退伍军人中常见的苦难，而治疗很少。中风经常因血液脑屏障（BBB）的破坏而复杂化，从而导致脑肿胀和出血。在这个项目中，我们建议研究中风后大脑中增加的蛋白质的潜在有益作用，HSP70。 HSP70通常不存在于大脑中，除了诸如中风之类的压力条件下。但是，中风过程中的HSP70进一步增加可以减少随后的脑损伤的量，并防止中风并发症。我们建议探索HSP70如何防止脑肿胀和出血，以便增加HSP70或管理HSP70本身的药理学策略可能是人类中风的有用治疗方法。由于中风是退伍军人中常见的医疗问题，因此该提案的工作可能会导致受益于该人群的治疗。