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

基本信息

批准号：
10093151
负责人：
Judy Shih-Hwa Liu
金额：
$ 43.15万
依托单位：
BROWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-02-01 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10093151
关键词：
Acidic Amino Acids Biological Markers Brain Brain region Caring Cell Line Cells 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 protein complex 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

项目摘要

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：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，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育畸形、癫痫发作情况各不相同，并且有非癫痫源性 FCD 病例的报道。哪些因素引发 FCD 中的癫痫尚不清楚。然而，对于小儿癫痫，大多数治疗性切除都是进行的适用于患有严重难治性癫痫发作的 FCD 患者。从此程序中收集的组织允许我们研究这些皮质畸形的癫痫发生。发现分子途径并识别为了寻找癫痫的潜在治疗靶点，我们储存了切除手术并进行了转录组分析 FCD 和相关疾病结节性硬化症 (TSC) 的致癫痫组织。我们对顽固性局灶性癫痫手术的手术样本进行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）患者。这该研究中基因表达的统计分析发现，mRNA 水平下降转录因子，昼夜节律运动输出周期 Kaput（时钟），在致癫痫组织中的表达 FCD 和 TSC 均与非癫痫大脑进行比较。这一结果得到了西方分析的证实更大的 FCD 病例队列。我们发现兴奋性和抑制性神经元的时钟都减少了。我们通过选择性删除皮层兴奋性神经元或抑制性神经元中的时钟创建了小鼠品系神经元。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。基于这些结果，我们假设时钟的丢失会破坏下游基因表达，从而导致电路功能障碍和癫痫。相反，维护时钟功能可以防止电路和分子引起癫痫的变化。我们将从三个目标来检验这个假设： 1) 我们将确定时钟功能丧失对皮质微电路的影响。 2）我们将确定一个通过研究其靶标 PARbZip 转录因子，了解时钟的分子机制。 3）我们将使用小昼夜节律转录遗传技术的分子修饰剂可挽救时钟减少的影响。这方法有可能通过开发新的治疗策略和改进来改善癫痫护理癫痫生物标志物。