Fas-Mediated Cell Death in TBI

TBI 中 Fas 介导的细胞死亡

• 批准号: 8097958
• 负责人: STEVEN H GRAHAM
• 金额: $ 14.73万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8097958
• 关键词: Address; Amyloid; Apoptosis; Behavioral; Binding; Brain; CD95 Antigens; Caspase; Cell Death; Cell Surface Receptors; Cells; Cessation of life; Chimeric Proteins; Cleaved cell; Complex; Cytosol; Data; Deposition; Dominant-Negative Mutation; Event; Functional disorder; Gene Family; Genes; Human; Infusion procedures; Injury; Laboratories; Mediating; Membrane Proteins; Middle Cerebral Artery Occlusion; Mitochondria; Molecular; Mus; Neurons; Outcome; Pathogenesis; Pathologic; Pathway interactions; Permeability; Physiological Processes; Principal Investigator; Proteins; Regulator Genes; Rodent Model; Role; Sampling; Site; Specimen; Stimulus; TBI Patients; Testing; Therapeutic Intervention; Trauma; Traumatic Brain Injury; Tumor Necrosis Factor Ligand Superfamily Member 6; Viral; Viral Vector; Virus Inhibitors; adeno-associated viral vector; base; caspase-3; caspase-8; caspase-9; cell type; controlled cortical impact; cytochrome c; functional outcomes; human disease; in vivo; inhibitor/antagonist; injured; loss of function mutation; neuron apoptosis; neuron loss; novel; overexpression; prevent; pro-caspase-8; programs; receptor

One molecule known to initiate programmed cell death in a variety of cell types in normal and pathologic conditions is the cell-surface receptor Fas. Fas is activated by another membrane protein, the Fas-ligand (FasL). Data from our and other laboratories demonstrate that Fas expression is increased and caspase-8 is cleaved and activated after trauma both in rodent models and in human TBI specimens. We have used two endogenous molecules that inhibit Fas-mediated cell death, Bcl-xL and cFLIP-L, to develop novel means of inhibiting Fas-mediated cell death. Therefore, the hypothesis to betested is that Fasis activated by FasL after trauma and initiates programmed cell death of neurons and worsens functional outcome. The following specific aims will be addressed: Aim 1. Test whether FasL function contributes to neuronal death and behavioral dysfunction after controlled cortical impact (CCI) in vivo by testing mice with a loss-of-function mutation in FasL (gld). Determine how the loss of FasL function effects the downstream events that execute programmed cell death afterCCI. Aim 2. Test whether the Fas pathway contributes to neuronal death after CCI in vivo by overexpression of cFLIP-L, a dominant negative inhibitor of Fas-mediated cell death, using an adeno- associated viral (AAV) vector. Aim 3. Test whether overexpression of Bcl-xL in neurons by systemic infusion of Bcl-xL-TAT fusion protein prior to and after injury protects mouse brain againstCCI. Aim 4. Determine if changes in Fas,FasL, Bid and caspase-8 occur in brain and CSF samples from patients with TBI and if these changes correlate with long-term outcome. Determine if increased expression of Fas is associated with (3-amyloid (Af3) deposition. The broad long-term objective of this project is to elucidate the molecular mechanisms of neuronal cell death after trauma by studying the role of the cell-death receptor Fas and related death-regulated gene products. Thus, this project may contribute to a better understanding of not only the mechanisms of neuronal cell death in TBI, but also suggest new treatment approaches.

已知一种分子可在正常和异常的多种细胞类型中启动程序性细胞死亡 病理条件下是细胞表面受体Fas。Fas被另一种膜蛋白激活， Fas配体（FasL）。我们和其他实验室的数据表明，Fas表达增加， 在啮齿动物模型和人TBI样本中，半胱天冬酶-8在创伤后被切割和活化。我们 已经使用两种抑制Fas介导的细胞死亡的内源性分子Bcl-xL和cFLIP-L来开发 抑制Fas介导的细胞死亡的新方法。 因此，有待验证的假设是，创伤后FasL激活Fas，并启动 神经元的程序性细胞死亡和神经元的功能结果。 将处理以下具体目标： 目标1.测试FasL功能是否有助于神经元死亡和行为功能障碍， 通过测试具有FasL（gld）功能缺失突变的小鼠，在体内控制皮质撞击（CCI）。 确定FasL功能的丧失如何影响执行程序性细胞死亡的下游事件 在CCI之后。 目标2.通过以下方法检测Fas通路是否有助于体内CCI后的神经元死亡： cFLIP-L是Fas介导的细胞死亡的显性负性抑制剂， 相关病毒（AAV）载体。 目标3.通过全身输注Bcl-xL-达特融合蛋白检测Bcl-xL是否在神经元中过表达 蛋白质在损伤前后保护小鼠脑免受CCI。 目标4。确定Fas、FasL、Bid和半胱天冬酶-8的变化是否发生在脑和CSF样品中， TBI患者以及这些变化是否与长期结果相关。确定是否增加表达 Fas的表达与β-淀粉样蛋白（Af 3）的沉积有关。 这个项目的广泛的长期目标是阐明神经元的分子机制， 通过研究细胞死亡受体Fas及相关死亡调控基因在创伤后细胞死亡中的作用 产品.因此，该项目不仅有助于更好地理解神经元的机制， TBI中的细胞死亡，但也提出了新的治疗方法。