Persisting Grand Mal and Myoclonic Seizures in JME including Veterans

JME 中持续出现大发作和肌阵挛发作，包括退伍军人

• 批准号: 8680017
• 负责人: ANTONIO V. DELGADO-ESCUETA
• 金额: --
• 依托单位: VA GREATER LOS ANGELES HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2017-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8680017
• 关键词: Affect; Antiepileptic Agents; Apoptosis; Area; Binding; Binding Sites; Brain Stem; Brazil; Cell division; Cells; Clinic; Clinical; Corpus striatum structure; Crystallography; Dendrites; Development; Developmental Gene; Disease; Dorsal; Drosophila genus; Drosophila melanogaster; Enrollment; Epilepsy; Epileptogenesis; Etiology; Genes; Genetic; Genome; Globus Pallidus; Grant; Growth; Health; Healthcare Systems; Human; Intestines; Ions; Juvenile Myoclonic Epilepsy; Knock-out; Lead; Los Angeles; Medicine; Microtubules; Mission; Modeling; Mus; Mutate; Mutation; Mutation Analysis; Myoclonic Epilepsies; Organized by Structure Protein; Pathway interactions; Patients; Peru; Phosphotransferases; Pontine structure; Population; Post-Traumatic Epilepsy; Predisposition; Process; Protein Binding; Proteins; Public Health; Recruitment Activity; Role; Services; Soldier; Staging; Substantia nigra structure; Symptomatic Generalized Epilepsy; Synapses; System; Testing; Thalamic structure; Tonic - clonic seizures; Traumatic Brain Injury; United States National Institutes of Health; Veterans; Work; base; cohort; deep sequencing; deletion analysis; functional gain; in vivo; indexing; member; migration; mouse model; mutant; neuroblast; neuropathology; next generation sequencing; novel; public health relevance; putamen; research study; screening

DESCRIPTION (provided by applicant): Epilepsies affect about 3 million people in the US and 70 million in the world. In Greater Los Angeles area there are 8,500 Veterans with epilepsy. The VA healthcare system is challenged by 300,000 to 400,000 USA soldiers returning from OEF/OIF; 212,000 service members suffering traumatic brain injury (TBI) over the last decade; and some 9,600 Veterans are already under treatment for post-traumatic epilepsy (PTE). The specific role of heritable epilepsies in PTE remains unresolved. Amongst the heritable epilepsies, Juvenile Myoclonic Epilepsy (JME) is the most common and there is no cure. To find a cure, we have been expanding the JME genome, increasing our understanding of the functions of myoclonin1/EFHC1, a JME causing gene, and defining new putative major JME gene(s) by next generation sequencing. Now propose to continue studying the myoclonin1/EFHC1 disease mechanisms of persisting grand mal and myoclonic seizures in JME. We will also study how important new putative epilepsy genes are in clinic based and Veterans cohort. If a putative epilepsy gene is in at least 3% of our cohorts, we seek proof for causality by replication of epilepsy in knockout (Ko) or knockin (Ki) mice model. The results guide which putative epilepsy genes have priority for studies of functions and disease mechanisms in Drosophila melanogaster (Aim 3), mice (Aim 2) and humans (Aim 1). Specific Aim 1 involves human mutation analysis of JME genes and asks: Are mutations in Myoclonin1/EFHC1 population specific? (Aim 1a) Screen for new mutations in myoclonin1/EFHC1 in 442 recruited/available human JME index cases (65 from Peru, 120 from Brazil, 137 from LA, and 100 Veterans in LA) and 1,000 controls and confirm the two stages of epilepsy development: (a) susceptibility/epileptogenicity, and (b) established epileptogenesis. [Note: Human material is available from our complementary NIH grant and this project will only recruit/enroll Veterans.] (Aim1b) Screen for mutations in new putative JME genes being identified by the epilepsy deep sequencing pipeline (our complementary NIH grant). Specific Aim 2 proves causality by knockout of JME gene in mice. (Aim 2a) Conduct neuropathological analysis of the striatum-nigro-thalamic-cortical and pedunculopontine pathway system, and compare neuropathology of caudate, putamen, globus pallidus, thalamus substantia nigra and brainstem especially superior coliculus and dorsal pons varolii of Myoclonin1/efhc1 KO mice with myoclonias only versus Myoclonin1/efhc1 KO mice with grand mal seizures. (Aim 2b) Develop a KO mouse of new putative JME genes that satisfy the screening process of the epilepsy deep sequencing pipeline. Specific Aim 3 studies the Drosophila model. (Aim 3a) Analyze in vivo how pathological mutations impact Drosophila EFHC1 protein (Defhc1.1) role in synaptic and dendritic development. (Aim 3b) Characterize Defhc1.2 mutant as well as the double Defhc1.1/Defhc1.2 mutant. (Aim 3c) Dissect Defhc1.1 by deletion analysis to identify the portion of the protein that binds to microtubules, and confirm that it is also the same binding sit in Defhc1.2. (Aim 3d) Because EFHC1 proteins bind microtubules through a non-canonical microtubule binding domain, a 'pilot experiment' will determine the structure of the protein by crystallography. (Aim 3e) Exploit the Drosophila model and gain functional information on the new putative JME genes being identified through the epilepsy deep sequencing pipeline. Finally, until recently, common genetic epilepsies were thought to be 'ion channelopathies'. Now, we provide more proof that JME is due to mutations in developmental genes, like Myoclonin1/EFHC1, involved in cell division, apoptosis, neuroblast migration, and synapse and dendrite growth. Understanding JME disease mechanisms brings an era of targeted and designer medicine, and positively impacts the mission of the VA healthcare system and the health of Veterans.

描述（由申请人提供）： 癫痫病影响美国约300万人，全球7000万人影响。在大洛杉矶地区，有8,500名退伍军人患有癫痫病。 VA医疗保健系统受到了30万至40万美国士兵的挑战，这些士兵从OEF/OIF返回；在过去的十年中，有212,000名服务成员遭受创伤性脑损伤（TBI）；大约9,600名退伍军人已经接受了创伤后癫痫（PTE）的治疗。可遗传性癫痫在PTE中的具体作用尚未解决。在可遗传的癫痫中，少年肌阵挛性癫痫（JME）是最常见的，无法治愈。为了找到治愈方法，我们一直在扩展JME基因组，增进了对Myoclonin1/efhc1的功能的理解，这是一种引起基因的JME，并通过下一代测序定义了新的推定主要JME基因。现在建议继续研究JME中持续的大玛尔和肌阵挛性癫痫发作的Myoclonin1/EFHC1疾病机制。我们还将研究新的假定癫痫基因在临床和退伍军人队列中的重要性。如果假定的癫痫基因至少在我们的同类群中，我们通过在敲除（KO）或敲除（KI）小鼠模型中复制癫痫病来寻求因果关系证明。假定癫痫基因在果蝇中（AIM 3），小鼠（AIM 2）和人类（AIM 1）的功能和疾病机制研究的结果指南具有优先级。特定目的1涉及JME基因的人类突变分析，并提出：Myoclonin1/efhc1种群中的突变是否特定？ （AIM 1A）在442份招募/可用的人类JME指数案例中，在442个肌昂酸酯/EFHC1中进行新突变的屏幕（65个来自秘鲁，来自巴西的120个，洛杉矶的137，洛杉矶的137名，洛杉矶的100名退伍军人）和1,000个控件，并确认癫痫发育的阶段：（A）易感性/ePiLeptity and Epileptitigy and Epilepticity和（B） [注意：我们的补充NIH赠款可从我们的材料中获得，该项目只能招募/招募退伍军人。]（AIM1B）屏幕屏幕在新推定的JME基因中被癫痫的深层测序管道（我们的补充NIH赠款）鉴定出来。特定的目标2证明了小鼠JME基因的敲除。 (Aim 2a) Conduct neuropathological analysis of the striatum-nigro-thalamic-cortical and pedunculopontine pathway system, and compare neuropathology of caudate, putamen, globus pallidus, thalamus substantia nigra and brainstem especially superior coliculus and dorsal pons varolii of Myoclonin1/efhc1 KO mice with myoclonias only versus Myoclonin1/efhc1 ko小鼠，癫痫发作。 （AIM 2B）开发了一种新的假定JME基因的KO小鼠，该基因满足癫痫深层测序管道的筛选过程。特定目标3研究果蝇模型。 （AIM 3A）在体内分析病理突变如何影响果蝇EFHC1蛋白（DEFHC1.1）在突触和树突发展中的作用。 （AIM 3B）表征DEDHC1.2突变体以及双DEDHC1.1/DEDHC1.2突变体。 （AIM 3C）通过缺失分析解剖DEDHC1.1，以识别与微管结合的蛋白质的一部分，并确认它在DEFHC1.2中也是相同的结合位置。 （AIM 3D）由于EFHC1蛋白通过非典型的微管结合结构域结合微管，因此“初步实验”将通过晶体学确定蛋白质的结构。 （AIM 3E）利用果蝇模型并获得有关通过癫痫深层测序管道鉴定的新推定JME基因的功能信息。最后，直到最近，常见的遗传癫痫被认为是“离子通道病”。现在，我们提供了更多的证据，即JME是由于发育基因中的突变，例如Myoclonin1/efhc1，涉及细胞分裂，细胞凋亡，神经细胞迁移，突触和树突生长。了解JME疾病机制带来了一个有针对性和设计师医学的时代，并积极影响VA医疗保健系统和退伍军人的健康。