NEUROPROTECTIVE GROWTH FACTORS IN TRAUMATIC BRAIN INJURY

创伤性脑损伤中的神经保护生长因子

• 批准号: 6819711
• 负责人: DEBORAH J WATSON
• 金额: $ 31.7万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-12-01 至 2006-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6819711
• 关键词: acute disease /disorder; apoptosis; biological signal transduction; biotechnology; biotherapeutic agent; brain disorder chemotherapy; brain injury; chronic brain damage; cognition; disease /disorder prevention /control; disease /therapy duration; fibroblast growth factor; growth factor; immunocytochemistry; laboratory rat; laser capture microdissection; mitogen activated protein kinase; molecular pathology; nerve growth factors; neural degeneration; neural plasticity; neuromuscular system; neuroprotectants; nonhuman therapy evaluation; sensorimotor system; serial analysis of gene expression

DESCRIPTION: (Verbatim from the Applicant's Abstract) The mechanism(s) of delayed secondary injury following traumatic brain injury (TBI) may involve the alteration of specific intracellular signaling pathways involving the mitogen activated protein (MAP) kinases JNK and Erk 1/2, as well as long-term genomic changes. These pathogenic molecular events provide targets for treatment with growth factors intended to prevent or limit neurologic disability. The investigators, among others, have generated preliminary data suggesting that acute (24 hr postinjury) administration of exogenous NGF or basic fibroblast growth factor (bFGF) may be neuroprotective in experimental models of brain injury. Specific Aim 1 will assess whether administration of either NGF or bFGF in the acute post-traumatic period (beginning 24 hr postinjury) will attenuate long-term neurologic disability and neurodegenerative (apoptotic/necrotic) cell loss following experimental lateral fluid percussion (FP) brain injury in rats. The investigators will selectively infuse these growth factors directly into the injured brain over a 2-week postinjury period, and behaviorally evaluate the animals over a chronic 3-month postinjury period for neurologic motor and sensorimotor dysfunction, cognitive deficits, and regional cell death. Specific Aim 2 will evaluate the temporal and regional alterations in specific MAP kinase (JNK/Erk 1/2) signaling pathways following experimental FP brain injury, and relate the neuroprotective effects of therapy with NGF or bFGF to a reversal of the pathologic changes in JNK/Erk induced by trauma. They will use Western blotting and immunohistochemistry to support their pilot data that TBI results in activation of the pro-death kinase JNK signaling pathway and a concomitant decrease in signaling through the pro-survival Erk 1/2 pathway. The effects of NGF or bFGF therapy and cessation of 2-week therapy (i.e., growth factor withdrawal) on these trauma-induced signal transduction cascades will then be regionally and temporally evaluated. In Specific Aim 3, they will quantify the effects of NGF or bFGF therapy on trauma-induced alterations in expression profiles of genes involved in cell death/survival and long-term plasticity/remodeling of the injured CNS. The investigators will use laser capture microdissection and reverse-Northern hybridization techniques with custom-designed slot-blots to quantify expression levels of a selected panel of genes, including cell death/survival genes, cytoskeletal genes, growth-related proteins, and cytokines. Genomic changes will be confirmed at a translational level with immunohistochemistry for selected proteins. In Specific Aim 4, the investigators will selectively infuse NGF or bFGF into the injured brain over a 2-week treatment period beginning 2 weeks or 1 month postinjury and evaluate the ability of these delayed treatment paradigms to improve long-term neurological motor and cognitive function (up to 3 months postinjury) and attenuate progressive post-traumatic cell death.

描述：（逐字来自申请人的摘要） 创伤性脑损伤（TBI）后的迟发性继发性损伤可能涉及 涉及有丝分裂原的特异性细胞内信号通路的改变 活化蛋白（MAP）激酶JNK和Erk 1/2，以及长期基因组 变化这些致病性分子事件提供了用以下药物治疗的靶点： 旨在预防或限制神经功能障碍的生长因子。的 调查人员，除其他外，已经产生了初步数据表明， 急性（损伤后24小时）给予外源性NGF或碱性成纤维细胞 在实验性脑损伤模型中，生长因子（bFGF）可能具有神经保护作用 损伤具体目标1将评估是否给予NGF或bFGF 在急性创伤后阶段（伤后24小时开始）， 长期神经功能障碍和神经退行性（凋亡/坏死）细胞 大鼠实验性侧向液压冲击（FP）脑损伤后的损失。 研究人员将选择性地将这些生长因子直接注入 在伤后2周内，对受伤的大脑进行行为评估， 在慢性损伤后3个月内，动物的神经运动和 感觉运动功能障碍、认知缺陷和局部细胞死亡。具体 目标2将评估特定MAP的时间和区域变化 实验性FP脑损伤后JNK/Erk 1/2信号通路， 并将神经生长因子或碱性成纤维细胞生长因子治疗的神经保护作用与 逆转由创伤引起的JNK/Erk的病理变化。他们将使用 免疫印迹和免疫组织化学来支持他们的试验数据， 导致促死亡激酶JNK信号通路的激活， 通过促存活Erk 1/2途径的信号传导的伴随减少。的 NGF或bFGF治疗的效果和2周治疗的停止（即，增长 因子撤除）对这些创伤诱导的信号转导级联的影响将 然后进行区域和时间评估。在第三个目标中，他们将 量化NGF或bFGF治疗对创伤诱导的改变的影响， 参与细胞死亡/存活和长期存活的基因的表达谱 受损CNS的可塑性/重塑。调查人员将使用激光 捕获显微切割和反向Northern杂交技术， 定制设计的狭缝印迹，以定量所选的一组 基因，包括细胞死亡/存活基因，细胞骨架基因，生长相关基因， 蛋白质和细胞因子。基因组变化将在翻译时确认。 免疫组化水平的选定蛋白质。具体目标4： 研究人员将选择性地将NGF或bFGF注入受伤的大脑， 2-从伤后2周或1个月开始的一周治疗期，并评估 这些延迟治疗范例改善长期 神经运动和认知功能（伤后3个月）， 减轻进行性创伤后细胞死亡。