Pathophysioloy of non-classic epileptic encephalopathies (EE)

癫痫性脑病（EE）是一种罕见但常见的中枢神经系统（CNS）疾病。EE是严重的、难治性癫痫，通常始于儿童期，并与认知结果不良相关。虽然癫痫性脑病可以作为CNS中可识别病变（如畸形）的结果发生，但在广泛的病例亚组中无法识别致病因素。遗传因素也有牵连，但直到最近，发现潜在的遗传改变才被证明是难以捉摸的。随着高通量测序技术的出现，可以确定经典癫痫性脑病的遗传基础，包括婴儿痉挛症（IS）和Lennox-Gastaut综合征（LGS）。发现新生突变对这些疾病的病因学有显著贡献，目前正在进行大型合作研究，以增加IS和LGS中解释的病例比例。在这些国际研究中，申请人已经作为项目负责人和协调员发挥了主导作用。在目前的建议中，我们的目标是扩大我们的基因识别策略，以更广泛的EE，如非经典的形式，不能归类为传统的综合征。这些EE在临床实践中构成了主要的诊断和治疗挑战，迄今为止尚未纳入系统研究。有强有力的证据表明，新生突变在这些疾病中起着重要作用，这些疾病具有高度的临床相关性，目前没有足够的治疗选择。我们的目标是在100例患者-父母三联体中使用三联体外显子组测序来鉴定从头突变，并在500例患者中使用50个最有希望的候选基因的基因组分析进行随访分析。我们将在5个最有希望的蛋白质中进行功能分析。鉴于在国家和欧洲范围内缺乏可比项目，本研究将为启动未来对迄今为止被忽视的广泛EE的确认和功能研究提供必要的基础。当前的提案申请是申请人在2012年12月重新提交的先前提案。我们认为，我们充分考虑了审查人员和DFG小组的意见和建议。此外，我们提出了以下方面的建议：（1）改变本申请某些方面的本领域的最新发展，以及（2）申请人专业知识的进展，特别是关于下一代测序技术。我们还强调了该申请在该领域整体竞争和资金格局中的重要性，因为该提案将使申请人能够确立其在该领域的领导者地位。

Epileptic encephalopathies (EE) are individually rare but collectively common disorders of the central nervous system (CNS). EE are severe, intractable epilepsies that usually start in childhood and are associated with a poor cognitive outcome. While epileptic encephalopathies can occur as the result of identifiable lesions in the CNS such as malformations, a causative factor cannot be identified in a broad subset of cases. Genetic factors are implicated, but until recently, the discovery of the underlying genetic alterations turned out to be elusive. With the advent of high-throughput sequencing technologies, a genetic basis could be identified for the classic epileptic encephalopathies including Infantile Spasms (IS) and the Lennox-Gastaut syndrome (LGS). De novo mutations were found to contribute significantly to the etiology of these conditions and large, collaborative studies are currently performed to increase the fractions of cases explained in IS and LGS. In these international studies, the applicants are already taking a leading role as project leaders and coordinators. In the current proposal, we aim to expand our gene-identification strategy to a broader range of EE such as the non-classical forms which cannot be classified into the traditional syndromes. These EE, which pose a major diagnostic and therapeutic challenge in clinical practice, have not been included in systematic studies so far. There is strong evidence that de novo mutations play a large role in these conditions, which are clinically highly relevant and currently without sufficient treatment options. We aim to identify de novo mutations using trio exome sequencing in 100 patient-parent trios with follow-up analysis in 500 patients using a gene panel analysis of the 50 most promising candidate genes. We will perform functional analysis in the 5 most promising proteins. Given the absence of comparable projects on a national and European scale, this study will provide the necessary foundation to launch future confirmation and functional studies on a broad range of so far neglected EE. This current proposal application is a resubmission of a previous proposal by the applicants in December 2012. We believe that we fully address the comments and suggestions of the reviewers and of the DFG panel. In addition, we put out proposal in context to (1) recent developments in the field that change some aspects of this application and (2) progress in the expertise of the applicants particularly with respect to next-generation sequencing technologies. We also address the importance of the application in the context of overall competition and funding landscape in this area, as this proposal will allow the applicants to establish their role as leaders in the field.