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.

项目概要/摘要 c-Jun N-末端激酶（JNK）是蛋白激酶家族，其是蛋白激酶家族的成员。 更大类别的“应激激活蛋白激酶”。已知JNK在细胞内被过度激活， 神经退行性疾病，如阿尔茨海默病，帕金森病，还有更多 最近与难治性癫痫有关在这些疾病中，JNK的过度活化是 这是一种病理事件，似乎有助于神经元的进行性凋亡性死亡。我们 最近发现，有些出乎意料的是，JNK在一种动物模型中被过度激活， 颞叶癫痫和JNK部分药理学抑制产生了 慢性癫痫患者自发性发作频率的显著剂量依赖性降低 动物此外，在三种JNK亚型（JNK 1、2和3）中，只有JNK 2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JNK 2可以产生一种抑制剂。 显著抗癫痫作用和避免广谱JNK潜在的CNS副作用 抑制作用 在这个为期两年的重点提案中，我们计划进一步研究JNK作为一个新目标， 抗癫痫治疗在第一个目标中，我们将寻求更完全地敲除JNK 2 使用新的AAV递送的CRISPR/Cas9在癫痫动物模型中的表达 approach.已发表的数据表明，这种方法能够敲低基因表达， 达到与组成型基因缺失相当的程度。我们将测量近- 在颞叶癫痫动物模型中完全敲除JNK 2对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JNK在人类癫痫中过度激活的证据， 在我们的动物模型中，通过寻找总的和磷酸化的（激活的）JNK表达， 对于接受脑切除术的人类癫痫患者的脑组织中的所有三种亚型， 难治性癫痫我们还将测量JNK的表达和激活， 难治性癫痫患者的脑脊液与没有癫痫症状的个体相比， 神经紊乱 这些实验将进一步验证JNK作为抗癫痫的新靶点 疗法迫切需要这样的目标，以推进对患有 难治性癫痫