Temporal Lobe Epilepsy and Retrotransposons

颞叶癫痫和逆转录转座子

基本信息

批准号：
9064579
负责人：
Wade H Berrettini
金额：
$ 25.05万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-30 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9064579
关键词：
Address Affect Antibodies Anticonvulsants Brain Brain Diseases Cell Nucleus Cell Separation Cell physiology Central Nervous System Diseases Chromatin Structure Chronic Chronic Disease Cloning Complex Craniotomy Cryptogenic Epilepsies DNA DNA Sequence Data Daughter Detection Development Disease Electroencephalography Elements Embryonic Development Epilepsy Event Excision Family Study Fluorescence Gene Expression Gene Frequency Genes Genetic Genetic Risk Genetic Transcription Genome Genomic Segment Goals High-Throughput Nucleotide Sequencing Human Genome Image Infection Inherited Introns Junk DNA L1 Elements Label Mediating Morbidity - disease rate Mosaicism Mutation Neuronal Differentiation Neurons Nuclear Organism Partial Epilepsies Paste substance Patients Pharmaceutical Preparations Pharmacotherapy Play Population Procedures Public Health RNA RNA Polymerase II RNA-Directed DNA Polymerase Regimen Retrotransposon Risk Role Sampling Secondary to Seizures Sequence Alignment Site Staining method Stains Temporal Lobe Temporal Lobe Epilepsy Testing Therapeutic Tissues Traumatic CNS injury Twin Studies Variant base brain cell case control digital endonuclease improved malformation mortality public health relevance risk variant success transposon/insertion element tumor

Address Affect Antibodies Anticonvulsants Brain Brain Diseases Cell Nucleus Cell Separation Cell physiology Central Nervous System Diseases Chromatin Structure Chronic Chronic Disease Cloning Complex Craniotomy Cryptogenic Epilepsies DNA DNA Sequence Data Daughter Detection Development Disease Electroencephalography Elements Embryonic Development Epilepsy Event Excision Family Study Fluorescence Gene Expression Gene Frequency Genes Genetic Genetic Risk Genetic Transcription Genome Genomic Segment Goals High-Throughput Nucleotide Sequencing Human Genome Image Infection Inherited Introns Junk DNA L1 Elements Label Mediating Morbidity - disease rate Mosaicism Mutation Neuronal Differentiation Neurons Nuclear Organism Partial Epilepsies Paste substance Patients Pharmaceutical Preparations Pharmacotherapy Play Population Procedures Public Health RNA RNA Polymerase II RNA-Directed DNA Polymerase Regimen Retrotransposon Risk Role Sampling Secondary to Seizures Sequence Alignment Site Staining method Stains Temporal Lobe Temporal Lobe Epilepsy Testing Therapeutic Tissues Traumatic CNS injury Twin Studies Variant base brain cell case control digital endonuclease improved malformation mortality public health relevance risk variant success transposon/insertion element tumor

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项目摘要

DESCRIPTION (provided by applicant): Idiopathic (cryptogenic) epilepsy is a common, chronic group of brain disorders, affecting 1-2% of the US population, creating a significant public health problem because of the associated morbidity and mortality. One subtype of this complex group of brain disorders, temporal lobe epilepsy (TLE), is particularly burdensome, because a substantial fraction of patients do not respond to anticonvulsant medications. If the seizure focus can be localized by imaging and/or EEG, the TLE patient often elects to have a partial resection of the affected temporal lobe, because they have daily seizures, despite multiple pharmacotherapy regimens. Although genetic risk is thought to play a role in TLE, identification of common risk alleles has had limited success. In the past 5 years, data have accumulated to prove that neuronal embryogenesis is accompanied by activation of LINE1 (L1) retrotransposons (RTPs) to an unexpected degree, such that developing neurons may accumulate de novo L1s insertions. This results in a substantial mosaicism within populations of CNS neurons. While most of these somatic de novo L1s will have little effect on neuronal function (perhaps because they occur in gene deserts or in large introns or in genes not required for that cell's function), some may interfere with normal neuronal activity because they have inserted into a gene needed by that particular neuron for normal function. If one or more functional de novo L1s occur early in CNS development, all the daughter neurons that derive from that neuronal precursor will also carry the L1 insertion, perhaps leading to a dysfunctional population of neurons destined to become an epileptic focus. This proposal will leverage CNS tissue (obtained at craniotomy) from patients with intractable TLE, who have undergone a therapeutic partial resection of one temporal lobe. Using fluorescence-assisted cell sorting of temporal lobe neuronal nuclei, followed by high-throughput sequencing and alignment to the reference genome, detection of de novo L1s will be done. The allele frequencies in neuronal DNA of selected de novo intragenic L1s will be determined by droplet digital PCR. In this manner, it is expected that de novo intragenic L1s, contributing to idiopathic TLE, will be discovered.

描述（由应用程序提供）：特发性（隐性）癫痫是一种常见的慢性脑疾病群，影响了1-2％的美国人群，由于相关的发病率和死亡率而造成了重大的公共卫生问题。这一复杂的脑疾病临时叶癫痫（TLE）的一种亚型特别繁重，因为很大一部分患者对抗惊厥药不反应。癫痫发作的焦点可以通过成像和/或脑电图定位，TLE患者通常会选择对受影响的临时叶进行部分切除，因为它们每天癫痫发作，进行多种药物治疗方案。尽管遗传风险被认为在TLE中起作用，但对共同风险等位基因的识别取得了有限的成功。在过去的5年中，数据积累以证明神经元的胚胎发生是通过激活Line1（L1）反转座子（RTP）（RTP）达到意外程度的，因此发展神经元可能会积累从头l1s插入。这导致了中枢神经系统神经元种群中的大量镶嵌。尽管这些从头l1的大多数对神经元功能的影响很小（也许是因为它们发生在基因沙漠，大型内含子中或该细胞功能不需要的基因中），但有些人可能会干扰正常的神经元活性，因为它们已插入该特定神经元所需的基因中，以实现正常功能。如果从中枢神经系统发育的早期出现一个或多个功能性的L1，那么从该神经元前体衍生的所有女儿神经元也将带来L1插入，也许会导致有可能成为癫痫焦点的神经元功能障碍。该提案将利用可顽固的TLE患者的CNS组织（在颅骨切开术时获得），他们使用荧光辅助的临时Lobe神经元核的细胞分选进行了A，然后对参考基因组进行高通量测序，然后对NOVO L1S的参考基因组进行对齐，将确定De Novo L1S的检测。通过这种方式，可以发现从头开始的基因内L1s，从而促成特发性TLE。