Targeting cellular senescence to prevent epileptogenesis

项目摘要 癫痫是一种以反复发作为特征的慢性神经疾病，影响6500万人。 并被认为是几种神经退行性疾病的共病，包括阿尔茨海默病(AD)。 确定目标以防止癫痫的发展(即癫痫的发生)是一个巨大的未得到满足的需求。 抗癫痫治疗仍然有限，即使在临床前模型中也是如此。在这里，我们建议研究一部小说 靶点是大脑中的衰老细胞(SCs)，以确定消融这些细胞是否可以防止癫痫的发生 匹罗卡品致小鼠颞叶癫痫模型及APdE9小鼠AD模型的研究 癫痫。细胞衰老是一种保守的细胞程序，可以阻止增殖并触发促细胞衰老。 炎性衰老相关分泌表型(SASP)对损伤性刺激的反应。公务员事务局局长是 人们对神经退行性疾病的兴趣与日俱增，但在癫痫的背景下仍然完全没有被探索。 致痫的侮辱，如癫痫持续状态(SE)，强烈地触发DNA损伤，这是细胞的一种常见原因 衰老。此外，在癫痫发生过程中报告的促炎细胞因子的特征也很惊人。 与SASP相似。我们将使用INK-ATTAC小鼠模型，它选择性地表达可诱导的 P16启动子控制下的自杀基因(和GFP)；p16的表达是SCs的标志。在目标1中， 我们将首先确定匹罗卡品诱导的SE后p16阳性细胞的细胞特性(胶质细胞和神经元细胞)。 接下来，我们将确定在SE后，去除SCs是否会减少促炎细胞因子的表达。最后，我们会 确定消融SCs是否减少了自发性癫痫的发生和/或减少了 一旦发作就会发作。在目标2中，我们将针对具有不同病因的APdE9小鼠模型。 AD显示共病癫痫发作和SCs积聚。与目标1中一样，我们将首先描述 APdE9模型中干细胞的细胞表型。接下来，我们将确定切除干细胞是否能减少促炎作用。 APdE9小鼠体内细胞因子水平。最后，我们将确定切除干细胞是否能减少自发性癫痫发作。 发生在APdE9小鼠身上。通过使用两种不同的病因和临床表现的模型 (匹罗卡品后的惊厥发作，APdE9模型中的非惊厥发作)，我们准备测试 最重要的假设是，干细胞参与了自发性癫痫的出现 癫痫发生，与仅特定模型所特有的效应相比。细胞衰老是 因此，这些研究有可能开辟研究癫痫的新途径 癫痫发生的机制。

Project Abstract Epilepsy is a chronic neurological condition characterized by recurrent seizures, affecting 65 million individuals and is recognized as a comorbidity in several neurodegenerative diseases, including Alzheimer’s disease (AD). Identifying targets to prevent the development of epilepsy (i.e., epileptogenesis) is a large unmet need. Antiepileptogenic therapies remain limited, even in preclinical models. Here, we propose to examine a novel target, senescent cells (SCs) in the brain, to determine if ablating these cells can prevent epileptogenesis in the pilocarpine mouse model of temporal lobe epilepsy and in the APdE9 mouse model of AD with comorbid epilepsy. Cellular senescence is a conserved cellular program which halts proliferation and triggers a pro- inflammatory senescence associated secretory phenotype (SASP) in response to damaging stimuli. SCs are of growing interest in neurodegenerative diseases but remain completely unexplored in the context of epilepsy. Epileptogenic insults, such as status epilepticus (SE) robustly trigger DNA damage, a common cause of cellular senescence. Moreover, the pro-inflammatory cytokine profile reported during epileptogenesis shares striking similarities to the SASP. We will use the INK-ATTAC mouse model, which selectively expresses an inducible suicide gene (and GFP) under the control of the p16 promoter; p16 expression is a hallmark of SCs. In Aim 1, we will first determine the cellular identity (glial, neuronal) of p16 positive cells following pilocarpine-induced SE. We will next determine if ablating SCs reduces proinflammatory cytokine expression after SE. Finally, we will determine if ablating SCs reduces the development of spontaneous seizures and/or reduces expression of seizures once they occur. In Aim 2, we will target a model with a different etiology, the APdE9 mouse model of AD, which displays co-morbid seizures, and accumulation of SCs. As in Aim 1, we will first characterize the cellular phenotype of SCs in the APdE9 model. We will next determine if ablation of SCs reduces proinflammatory cytokine levels in APdE9 mice. Finally, we will determine if ablation of SCs reduces spontaneous seizure occurrence in APdE9 mice. By using two models, with very different etiologies and “clinical” presentation (convulsive seizures after pilocarpine, non-convulsive seizures in the APdE9 model), we are positioned to test the overarching hypothesis that SCs contribute to the emergence of spontaneous seizures during epileptogenesis, as compared to an effect that is specific to only a particular model. Cellular senescence is unexamined in epilepsy, thus, these studies have the potential to open new lines of research into the mechanisms of epileptogenesis.