Antiepileptic action of JNK2 inhibition

项目摘要/摘要 C-Jun氨基末端激酶(JNKs)是一类蛋白激酶家族，属于 更大类别的“应激激活蛋白激酶”。已知JNK在 神经退行性疾病，如阿尔茨海默病，帕金森氏病，以及 最近与难治性癫痫有牵连。在这些疾病中，JNK的过度激活是 这是一种病理事件，似乎与神经元的进行性凋亡死亡有关。我们 最近发现，有些出乎意料的是，在一种动物模型中，JNK过度激活 颞叶癫痫，JNK的部分药理抑制产生了 慢性癫痫患者自发性发作频率显著降低，呈剂量依赖性 动物。此外，在JNK的三个亚型(JNK1、2和3)中，只有JNK2被过度激活 在我们的动物模型中。因此，我们假设选择性抑制JNK2可以产生一种 广谱JNK显著的抗癫痫作用及避免潜在的中枢神经系统不良反应 抑制力。 在这项为期两年的重点计划中，我们计划进一步研究JNK作为 抗癫痫治疗。在第一个目标中，我们将寻求更全面地击倒JNK2 新型AAV载体CRISPR/Cas9在癫痫动物模型中的表达 接近。已公布的数据表明，这种方法能够抑制基因表达。 达到相当于结构性基因缺失的程度。我们将衡量近乎- 完全抑制JNK2基因对颞叶癫痫动物模型自发性癫痫发作频率的影响 脑叶癫痫。 在第二个目标中，我们将寻找证据表明JNKs在人类癫痫中被过度激活 在我们的动物模型中，它们通过寻找总的和磷酸化的(激活的)JNK表达来实现 人脑切除后癫痫患者脑组织中三种亚型的表达 医学上难治性癫痫。我们还将测量JNK的表达和激活 难治性癫痫患者脑脊液与非癫痫患者脑脊液的比较 神经性疾病。 这些实验将进一步证实JNK作为抗癫痫药物的新靶点。 心理治疗。这样的目标是迫切需要的，以推进对患有癌症的患者的治疗 医学上难治性癫痫。

Project summary / abstract c-Jun N-terminal kinases (JNKs) are a family of protein kinases that are members of the larger category of “stress activated protein kinases.” JNKs are known to be hyperactivated in neurodegenerative disorders such as Alzheimer’s disease, Parkinson’s disease, and are more recently implicated in medically refractory epilepsy. In these diseases, hyperactivation of JNK is a pathological event, and appears to contribute to the progressive apoptotic death of neurons. We recently found, somewhat unexpectedly, that JNK is hyperactivated in an animal model of temporal lobe epilepsy, and that partial pharmacological inhibition of JNK produced a significant, dose-dependent reduction of spontaneous seizure frequency in chronically epileptic animals. In addition, of the three JNK isoforms (JNK1, 2, and 3), only JNK2 was hyperactivated in our animal model. Thus, we hypothesized that selective inhibition of JNK2 could produce a significant antiepileptic action and avoid potential CNS adverse effects of broad spectrum JNK inhibition. In this two-year, focused proposal we plan to further study JNK as a novel target of antiepileptic therapy. In the first Aim, we will seek more complete knockdown of JNK2 expression in an animal model of epilepsy using a novel AAV-delivered CRISPR/Cas9 approach. Published data shows that this approach is capable of knocking down gene expression to a degree comparable to that of constitutive gene deletion. We will measure the effect of near- complete JNK2 knockdown on spontaneous seizure frequency in an animal model of temporal lobe epilepsy. In the second Aim, we will seek evidence that JNKs are hyperactivated in human epilepsy as they are in our animal model by looking for total and phosphorylated (activated) JNK expression for all three isoforms in brain tissue from human epilepsy patients undergoing brain resection for medically refractory epilepsy. We will also measure JNK expression and activation in cerebrospinal fluid from refractory epilepsy patients compared to individuals without a neurological disorder. These experiments will provide further validation of JNK as a novel target of antiepileptic therapy. Such targets are desperately needed in order to advance therapy for patients with medically refractory epilepsy.