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

科学摘要：局灶性皮质发育不良(FCD)是一种常见的异质性非遗传性疾病。 最常见的原因是难治性局灶性癫痫。而FCD大概是一种稳定的发展 畸形，癫痫发作是不同的，非致痫的FCD的病例被报告。是什么因素引发了 FCD中的癫痫是未知的。然而，在儿童癫痫中，大多数治疗性切除都是进行的。 适用于有严重药物难治性癫痫发作的FCD患者。从这个过程中收集的组织允许 我们将研究这些皮质畸形的癫痫发病机制。发现分子途径并鉴定 对于癫痫的潜在治疗靶点，我们将手术切除并进行了转录组分析 来自FCD和相关疾病结节性硬化症(TSC)的致痫组织。 难治性局灶性癫痫手术标本的初步转录组分析 病例包括局灶性皮质发育不良(FCD)和结节性硬化症(TSC)。这个 对该研究中基因表达的统计分析表明， 转录因子昼夜节律运动输出循环Kaput(CLOCK)，在致痫组织中的表达 FCD和TSC与非癫痫脑比较。这一结果得到了西方分析的证实 更大的FCD病例队列。我们发现兴奋性神经元和抑制性神经元的时钟都减少了。我们 建立了选择性缺失皮层兴奋性神经元或抑制性神经元时钟的小鼠系 神经元。我们发现，兴奋性神经元中时钟基因特异性缺失的小鼠具有自发性 癫痫发作。根据这些结果，我们假设生物钟的丢失破坏了下游基因的表达 到回路功能障碍和癫痫。相反，时钟功能的维持阻止了电路和分子 改变癫痫的致病因素。我们将从三个方面检验这一假设： 1)我们将确定时钟功能丧失对大脑皮质微电路的影响。2)我们将确定 通过研究其靶标PARbZip转录因子来研究Clock的分子机制。3)我们将使用Small 分子调节剂的昼夜节律转录遗传技术，以挽救时钟下降的影响。这 方法有可能通过开发新的治疗策略和改进来改善癫痫的护理 癫痫生物标志物。

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.