Antiepileptic action of JNK2 inhibition

JNK2 抑制的抗癫痫作用

• 批准号: 10515751
• 负责人: NICHOLAS P POOLOS
• 金额: $ 42.76万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10515751
• 关键词: Adverse effects; Alzheimer' s Disease; Animal Model; Animals; Antiepileptic Agents; Apoptotic; Astrocytes; Brain; CRISPR/Cas technology; Categories; Cerebrospinal Fluid; Chronic; Contralateral; Control Animal; Data; Disease; Dose; Electrodes; Epilepsy; Event; Excision; Frequencies; Gene Deletion; Gene Expression; Harvest; Hippocampus (Brain); Human; Immunohistochemistry; Impact Seizures; Implant; Individual; Interneurons; Intractable Epilepsy; KRP protein; MAP Kinase Gene; MAPK8 gene; MAPK9 gene; Maintenance; Measurement; Measures; Mediating; Medical; Microglia; Molecular; Monitor; N-terminal; Neuraxis; Neurodegenerative Disorders; Neurologic; Neurons; Parkinson Disease; Pathologic; Patients; Pharmacology; Phosphorylation; Phosphotransferases; Play; Population Control; Protein Family; Protein Isoforms; Protein Kinase; Proteins; Publishing; Rattus; Role; Sampling; Seizures; Side; Signal Pathway; Techniques; Temporal Lobe Epilepsy; Testing; Tissues; Validation; Work; acquired epilepsy; brain tissue; cell type; differential expression; experimental study; human tissue; innovation; knock-down; member; neocortical; nervous system disorder; neuron loss; novel; p38 Mitogen Activated Protein Kinase; stress activated protein kinase; tool

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.

项目摘要 /摘要 C-Jun N末端激酶（JNKS）是一个蛋白激酶家族，是 较大类别的“应激激活蛋白激酶”。已知JNK在 神经退行性疾病，例如阿尔茨海默氏病，帕金森氏病，更多 最近在医学性难治性癫痫中实施。在这些疾病中，JNK的过度激活是 一个病理事件，似乎有助于神经元的进行性凋亡死亡。我们 最近发现，有些人意外地，JNK在动物模型中被过度激活 临时叶癫痫，部分药物抑制JNK产生了 赞助商依赖性癫痫发作频率的显着，剂量依赖性降低 动物。此外，在三个JNK同工型（JNK1、2和3）中，只有JNK2被过度激活 在我们的动物模型中。那就是我们假设选择性抑制JNK2可以产生 明显的抗癫痫作用并避免了广谱JNK的潜在中枢神经系统不利影响 抑制。 在这个为期两年的集中建议中，我们计划进一步研究JNK作为新的目标 抗癫痫疗法。在第一个目的中，我们将寻求更全面的JNK2 使用新型AAV递送CRISPR/CAS9在癫痫动物模型中的表达 方法。已发表的数据表明，这种方法能够击倒基因表达 与本构基因缺失相当的程度。我们将衡量接近的效果 在临时动物模型中的赞助癫痫发作频率上完全JNK2敲低 叶癫痫。 在第二个目标中，我们将寻求证据证明JNK在人类癫痫中被过度激活为 它们是通过寻找全部和磷酸化（激活的）JNK表达的动物模型中的 对于来自接受大脑切除的人癫痫患者的所有三种同工型 医学上难治性癫痫。我们还将测量JNK表达和激活 与没有A的个体相比，来自难治性癫痫患者的脑脊液 神经系统障碍。 这些实验将进一步验证JNK作为抗胃静脉曲张的新目标 治疗。迫切需要这样的目标，以便为患者提供治疗 医学上难治性癫痫。