Modulating ProNGF-induced cell death in Epilepsy: strategies for neuroprotection

调节 ProNGF 诱导的癫痫细胞死亡：神经保护策略

基本信息

批准号：
8624717
负责人：
WILMA J FRIEDMAN
金额：
$ 47.74万
依托单位：
RUTGERS THE STATE UNIV OF NJ NEWARK
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-03-01 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8624717
关键词：
Acute Address Adult Alleles Animals Antibodies Apoptosis Astrocytes Attenuated Binding Brain C-terminal Cell Death Cells Cessation of life Chronic Cleaved cell Code Complex Detection Development Effectiveness Electroencephalography Elements Epilepsy Epileptogenesis Epitopes Equilibrium Exons FDA approved Genetic Hippocampus (Brain)Impaired cognition In Vitro Infusion procedures Injury Knock-in Mouse Lead Ligand Binding Ligands Mediating Modeling Mus NGFR Protein Nervous system structure Neuraxis Neuroglia Neuronal Differentiation Neuronal Injury Neurons Outcome Pattern Pharmaceutical Preparations Pilocarpine Preclinical Drug Evaluation Process Proteins Reagent Recurrence Relative (related person)Resistance Risk Rodent Model Role Seizures Signal Transduction Status Epilepticus Stroke Synapses Testing Toxic effect Traumatic Brain Injury Up-Regulation Wild Type Mouse cell type clinical application disability in vivo mouse model neuron apoptosis neuron loss neuronal survival neuroprotection neurotrophic factor neutralizing antibody novel patient population prevent promoter receptor sortilin tool transcriptional coactivator p75 uptake

项目摘要

DESCRIPTION (provided by applicant): Many types of insults or injury lead to seizures and neuronal loss, which ultimately can lead to devastating and intractable long-term consequences, including cognitive impairment, chronic epilepsy and disability. Our labs have identified a central role of neurotrophins, particularly proNGF in mediating neuronal apoptosis following severe seizures (status epilepticus; SE). Neurotrophins, initially characterized for their survival and differentiative actions on neurons, are initially synthesized as precursors that are cleaved to release C-terminal mature forms that bind to Trk receptors to promote neuronal survival, and synaptic actions. Recent studies suggest that the precursor form of NGF (proNGF) acts as a distinct ligand by binding to a receptor complex of p75 and sortilin to initiate neuronal death. Our labs have demonstrated that SE rapidly induces p75 and proNGF in the hippocampus, in rodent models. Importantly, infusion of function blocking anti-proNGF antibodies, or genetic deletion of p75, rescues neuronal death following SE, providing a mechanism and target for therapy to prevent neuronal loss after SE, and its consequence, epileptogenesis. Here we extend these collaborative studies to utilize novel knock-in mouse lines that expressed HA-epitope tagged NGF, or proNGF under its endogenous promoter elements. These animals circumvent the current limitations in sensitivity of reagents, and markedly enhance our ability to detect endogenous NGF. Thus we will be able to identify cell specific and temporal patterns of NGF/proNGF synthesis in neurons and glia under basal conditions and after SE. Using the proNGF mouse, we will quantitatively evaluate the effects of augmented proNGF following SE induction, to assess whether excess proNGF leads to exacerbated neuronal cell death. We will also use video-EEG to assess the consequence of excess proNGF on the development of epilepsy. Finally, and most important, we will investigate whether antagonists of proNGF, identified in a high through-put drug screen, can prevent or attenuate neuronal death following SE and prevent the onset of epilepsy. These antagonists consist of FDA-approved drugs that have minimal toxicity and efficiently block proNGF-induced apoptosis in cultured neurons. The possibility of identifying approved drugs that have the capacity to prevent neuronal death from severe seizures has significant potential for clinical application, both for epileptic patient populations, and for those suffering seizures as a consequence of brain trauma or stroke.

描述（由申请人提供）：许多类型的侮辱或伤害会导致癫痫发作和神经元损失，最终可能导致毁灭性且棘手的长期后果，包括认知障碍、慢性癫痫和残疾。我们的实验室已经确定了神经营养素，特别是 proNGF 在介导严重癫痫发作（癫痫持续状态；SE）后神经元凋亡中的核心作用。神经营养蛋白最初以其对神经元的存活和分化作用为特征，最初被合成为前体，被裂解释放出 C 端成熟形式，与 Trk 受体结合，促进神经元存活和突触活动。最近的研究表明，NGF 的前体形式 (proNGF) 作为一种独特的配体，通过与 p75 和分拣蛋白的受体复合物结合来启动神经元死亡。我们的实验室已经证明，SE 在啮齿动物模型的海马体中快速诱导 p75 和 proNGF。重要的是，输注功能阻断性抗 proNGF 抗体或 p75 基因删除可以挽救 SE 后的神经元死亡，为预防 SE 后神经元损失及其后果癫痫发生提供治疗机制和目标。在这里，我们扩展了这些合作研究，以利用在其内源启动子元件下表达HA表位标记的NGF或proNGF的新型敲入小鼠系。这些动物规避了目前试剂灵敏度的限制，并显着增强了我们检测内源性 NGF 的能力。因此，我们将能够识别基础条件下和 SE 后神经元和神经胶质细胞中 NGF/proNGF 合成的细胞特异性和时间模式。使用 proNGF 小鼠，我们将定量评估 SE 诱导后增强的 proNGF 的影响，以评估过量的 proNGF 是否会导致神经元细胞死亡加剧。我们还将使用视频脑电图来评估过量的 proNGF 对癫痫发展的影响。最后，也是最重要的，我们将研究在高通量药物筛选中鉴定出的 proNGF 拮抗剂是否可以预防或减轻 SE 后的神经元死亡并预防癫痫的发作。这些拮抗剂由 FDA 批准的药物组成，毒性最小，可有效阻断培养神经元中 proNGF 诱导的细胞凋亡。确定能够预防严重癫痫发作引起的神经元死亡的批准药物的可能性，对于癫痫患者群体以及因脑外伤或中风而遭受癫痫发作的患者来说具有巨大的临床应用潜力。