Cortical tuber and epilepsy

皮质结节和癫痫

• 批准号: 9053885
• 负责人: Angelique Bordey
• 金额: $ 25万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9053885
• 关键词: Address; Affect; Behavior; Cognitive; Complex; Cortical Dysplasia; Cortical Malformation; Data; Data Reporting; Dendrites; Development; Disease; Electroencephalogram; Electroporation; Embryo; Engineering; Epilepsy; Epileptogenesis; Excision; Experimental Models; Frequencies; Generalized seizures; Generations; Gliosis; Health; Human; Human Characteristics; Hyperactive behavior; Individual; Infantile spasms; Knock-out; Knockout Mice; Lead; Lesion; Medial; Modeling; Molecular; Monitor; Motor Seizures; Mutation; Nature; Neurologic; Neurons; Operative Surgical Procedures; Pacemakers; Pathology; Pathway interactions; Patients; Population; Positioning Attribute; Prefrontal Cortex; Procedures; Prosencephalon; Proteins; Refractory; Reporting; Seizures; Sirolimus; Slice; Symptoms; TSC1 gene; TSC1/2 gene; TSC2 gene; Tissues; Tuberous sclerosis protein complex; Tumor Suppressor Genes; Vigabatrin; Work; base; biophysical properties; body system; brain malformation; clinically relevant; cyclic-nucleotide gated ion channels; disability; experience; heart cell; human FRAP1 protein; improved; in utero; in vivo; innovation; insight; knock-down; malformation; novel; patch clamp; prevent; receptor; receptor coupling; research study; success; synaptic function; tool

 DESCRIPTION (provided by applicant): Tuberous sclerosis complex (TSC) is an autosomal dominant disorder occurring in 1/6,000 individuals with mutations in TSC1 or TSC2 leading to increased mTOR activity. About 80- 90% of the TSC individuals will experience seizures of different types. Most types of seizures are refractory to treatment and require invasive surgical resection leading to manageable seizures in only 50% of the cases. There is thus a serious need to improve epilepsy treatment in TSC. To develop such treatment, we need to better understand the mechanisms of seizure generation. It is known that focal cortical malformations, called cortical tubers in TSC are associated with epilepsy, but understanding how tubers generate seizures is limited because of the lack of an experimental model of focal cortical malformation-associated epilepsy. Here, we have generated a novel model of focal tuber-associated seizures, and we propose to further characterize it and address some of the possible mechanisms of epileptogenesis/seizures. As in humans, we found that experimental cortical tubers are characterized by loss of cortical lamination, the presence of ectopic, cytomegalic neurons with hypertrophic dendrites, and gliosis. Our overarching hypothesis is preventing one of the alterations may be sufficient to prevent seizure generation. Here, due to the exploratory nature and the short duration of a R21, we focused on two specific alterations, neuronal misplacement which is associated with circuit disorganization, and mTORC1-induced changes in biophysical properties (i.e., abnormal acquisition of pacemaker channel). Our more specific hypothesis is that preventing neuronal misplacement or specific alterations in biophysical properties prevents seizures. To address this hypothesis, we have two aims, one focused on neuronal misplacement and the other one on the biophysical properties of tuber neurons. We will use a combination of approaches including in utero electroporation to increase mTOR activity in developing cortical neurons followed by patch clamp recordings in slices and electroencephalogram (EEG)/behavior recordings to monitor epileptiform discharges in vivo. In addition, we will use conditional approaches as well as engineered receptors to silence neurons in vivo. This combination of approaches is innovative and we are in a unique position to accomplish the proposed work. Finally, TSC is an exemplary disorder of mTOR-dependent focal cortical malformation-associated epilepsy. The other disorder is focal cortical dysplasia (FCD) type II, which share similar molecular and cellular alterations. Our model is thus applicable to FCD type II. Both disorders are the leading cause of focal malformation-associated refractory epilepsy.s

 描述(申请人提供)：结节性硬化症(TSC)是一种常染色体显性遗传病，发生在1/6,000名TSC1或TSC2突变导致mTOR活性增加的患者中。大约80%-90%的TSC患者将经历不同类型的癫痫发作。大多数类型的癫痫发作难以治疗，需要侵入性手术切除，导致仅50%的病例癫痫发作可控。因此，迫切需要改进TSC的癫痫治疗。为了开发这种治疗方法，我们需要更好地了解癫痫发作的发生机制。众所周知，局灶性皮质畸形，在TSC中称为皮质结节与癫痫有关，但由于缺乏局灶性皮质畸形相关癫痫的实验模型，对结节如何产生癫痫的了解有限。在这里，我们建立了一个新的局灶性结节相关癫痫发作模型，我们建议进一步描述它的特征，并解决一些可能的癫痫发生/发作机制。正如在人类中一样，我们发现实验性皮质结节的特征是皮质板层丢失，存在异位的巨细胞神经元和肥大的树突，以及胶质细胞增生。我们的首要假设是防止其中一种改变可能足以防止癫痫的发生。在这里，由于R21的探索性和持续时间短，我们专注于两个特定的变化，与电路解体相关的神经元错位，以及mTORC1诱导的生物物理特性的变化(即起搏器通道的异常获取)。我们更具体的假设是，防止神经元错位或生物物理特性的特定变化可以防止癫痫发作。为了解决这一假设，我们有两个目标，一个集中在神经元错位上，另一个集中在块茎神经元的生物物理性质上。我们将结合使用多种方法，包括宫内电穿孔，以增加发育中的皮质神经元的mTOR活性，随后在脑片上进行膜片钳记录，并使用脑电/行为记录来监测体内的癫痫样放电。此外，我们将使用有条件的方法以及工程的受体来沉默体内的神经元。这种方法的结合是创新的，我们处于独特的地位，可以完成拟议的工作。最后，TSC是mTOR依赖的局灶性皮质畸形相关癫痫的典型障碍。另一种疾病是局灶性皮质发育不良(FCD)II型，具有相似的分子和细胞变化。因此，我们的模型适用于FCD II型。这两种疾病都是局灶性畸形相关难治性癫痫的主要原因。