The Circadian Molecular Clock is a Biomarker for Epilepsy in Focal Cortical Dysplasia

昼夜节律分子钟是局灶性皮质发育不良中癫痫的生物标志物

• 批准号: 10334417
• 负责人: Judy Shih-Hwa Liu
• 金额: $ 35.46万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10334417
• 关键词: Acidic Amino Acids; Biological Markers; Brain; Brain region; Caring; Cell Line; Cells; Circadian gene expression; Clinical; Cortical Dysplasia; Cortical Malformation; Data; Defect; Development; Disease; Electrophysiology (science); Enhancers; Epilepsy; Epileptogenesis; Excision; Exhibits; Frequencies; Functional disorder; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Techniques; Genetic Transcription; Goals; Human; Intractable Epilepsy; Lead; Magnetic Resonance Imaging; Maintenance; Measures; Mediating; Medical; Messenger RNA; Molecular; Molecular Profiling; Morbidity - disease rate; Mus; Neurons; Neurosurgeon; Operative Surgical Procedures; Output; Partial Epilepsies; Pathway interactions; Patients; Procedures; Proline; Proteins; Refractory; Reporting; Research; Sampling; Seizures; Severities; Statistical Data Interpretation; Testing; Therapeutic; Tissue Sample; Tissues; Tuberous Sclerosis; Validation; Vertebral column; bZIP Domain; base; brain abnormalities; brain tissue; childhood epilepsy; circadian; cohort; excitatory neuron; experimental study; hippocampal pyramidal neuron; human tissue; improved; inhibitory neuron; loss of function; malformation; molecular clock; mortality; mouse genetics; neuronal circuitry; novel marker; novel strategies; novel therapeutic intervention; optogenetics; patch clamp; personalized approach; postsynaptic; prevent; profiles in patients; restoration; side effect; small molecule; therapeutic target; transcription factor; transcriptome

Scientific Abstract: A common heterogeneous non-inherited condition, focal cortical dysplasia (FCD), is the most common cause of refractory focal epilepsy. While FCD is presumably a stable developmental malformation, epilepsy onset is variable and cases of non-epileptogenic FCD are reported. What factors initiate epilepsy in FCD are unknown. Nevertheless, in pediatric epilepsy most therapeutic resections are performed for FCD patients with severely medically refractory seizures. The tissue collected from this procedure allowed us to study epileptogenesis in these cortical malformations. To discover molecular pathways and to identify potential therapeutic targets in epilepsy, we banked resections and performed a transcriptome analysis of the epileptogenic tissue from FCD and a related disorder Tuberous Sclerosis Complex (TSC). Our preliminary transcriptome analysis on surgical samples from intractable focal epilepsy surgical cases included patients with focal cortical dysplasia (FCD) and tuberous sclerosis complex (TSC). The statistical analysis of gene expression in that study identified a decrease in the mRNA levels of the transcription factor, Circadian Locomotor Output Cycles Kaput (Clock), expression in epileptogenic tissue from both FCD and TSC compared with non-epileptic brain. This result was confirmed by Western analysis in a larger cohort of FCD cases. We found a reduction of Clock in both excitatory and inhibitory neurons. We created mouse lines with selective deletion of Clock in either excitatory neurons in the cortex or inhibitory neurons. We found that mice with specific deletion of the Clock gene in excitatory neurons have spontaneous seizures. Based on these results we hypothesize loss of Clock disrupts downstream gene expression leading to circuit dysfunction and epilepsy. Conversely, maintenance of Clock function prevents circuit and molecular changes causative for epilepsy. We will test this hypothesis in three aims: 1) We will determine the effect of Clock loss of function on cortical microcircuits. 2) We will determine a molecular mechanism for Clock by studying its targets, the PARbZip transcription factors. 3) We will use small molecule modifiers of circadian transcription genetic techniques to rescue the effects of decreased Clock. This approach has the potential to improve epilepsy care by developing new therapeutic strategies and refining epilepsy biomarkers.

科学摘要：一种常见的异质性非遗传性疾病，局灶性皮质发育不良（FCD），是 难治性局灶性癫痫的最常见原因。虽然FCD可能是一种稳定的发展， 畸形，癫痫发作是可变的和非致癫痫FCD的情况下，报告。哪些因素引发 FCD中的癫痫未知。然而，在小儿癫痫大多数治疗切除进行 对于患有严重医学难治性癫痫发作的FCD患者。从这个过程中收集的组织 我们来研究这些皮质畸形的癫痫发生。去发现分子通路， 作为癫痫的潜在治疗靶点，我们收集了切除标本，并对癫痫患者进行了转录组分析。 来自FCD的致癫痫组织和相关疾病多发性硬化症（TSC）。 我们对难治性局灶性癫痫手术标本的初步转录组分析 例为局灶性皮质发育不良（FCD）和结节性硬化症（TSC）患者。的 研究中基因表达统计分析表明， 转录因子，昼夜节律运动输出周期Kaput（时钟），在癫痫组织中的表达， FCD和TSC与非癫痫脑相比。这一结果得到了西方分析的证实， FCD病例的较大队列。我们发现在兴奋性和抑制性神经元中Clock减少。我们 创建了选择性删除皮质兴奋性神经元或抑制性神经元中Clock的小鼠品系。 神经元我们发现，在兴奋性神经元中特异性缺失Clock基因的小鼠， 癫痫发作。基于这些结果，我们假设Clock的丢失破坏了下游基因的表达， 电路功能障碍和癫痫相反，时钟功能的维护防止电路和分子 引起癫痫的变化。我们将在三个目标中检验这一假设： 1)我们将确定时钟功能丧失对皮层微电路的影响。2)我们将确定一个 通过研究Clock的靶点PARbZip转录因子来研究其分子机制。3)我们将使用小 分子修饰的昼夜转录遗传技术，以挽救减少时钟的影响。这 这种方法有可能通过开发新的治疗策略和改进治疗方法来改善癫痫护理。 癫痫生物标志物。