Mechanisms of PCDH19-clustering epilepsy

项目摘要/摘要 这份提案描述了一项为期五年的职业发展计划，该计划将引导PI成为一名 独立的医生-科学家，研究发育和疾病的机制和潜在的治疗方法 癫痫性脑病。 申请者：齐奥布罗博士拥有医学博士和博士学位，并完成了两个孩子的专业临床培训 神经病学与儿科癫痫。她以前有过使用啮齿动物进行神经科学研究的经验， 获得性癫痫的神经细胞培养和器官型海马细胞培养模型。这份职业 发展计划包括一段时间的指导性研究，旨在发展申请者在 先进的成像技术、病毒载体设计、脑切片电生理学和中间神经元发育。 这些技能将补充她目前的技能，并推动她作为一名独立研究员的发展。 在本培训期间学到的概念将广泛适用于多种疾病过程和 允许在与临床相关的研究领域进行重要的临床翻译。 研究计划：PCDH19-聚集性癫痫(PCE)是最常见的单基因发育 和癫痫脑病(DEE)，以认知障碍和难治性癫痫群为特征 从婴儿期开始。PCDH19是一种编码跨膜细胞黏附分子的X连锁基因， 对大脑发育过程中的细胞相互作用至关重要。PCE影响雌性和罕见的花叶雄性，而雄性 只表达突变的PCDH19不会导致癫痫。解释这一现象的一个主要假说是 它的发生是由于与随机X失活(或镶嵌突变)相关的细胞干扰 表达野生型和突变型PCDH19的细胞在脑内无法正常相互作用 发展，这得到了初步数据的支持，显示了Pcdh19+独特的细胞分离模式 以及PCE小鼠模型大脑皮质和海马区的Pcdh19-神经元。我们的中心假设是 嵌合体Pcdh19表达改变海马CA1区中间神经元发育导致异常 网络形成、过度兴奋和癫痫敏感性增加。这项提议将成为 PCE的组织学(目标1)、生理(目标2)和发育(目标3)机制。这项研究将提供 对PCE的机制有重要的洞察力并指导可能的潜在治疗策略 适用于多发性遗传性癫痫。

PROJECT SUMMARY/ABSTRACT This proposal describes a five-year career development program that will lead the PI to a career as an independent physician-scientist, studying mechanisms and potential therapeutics of developmental and epileptic encephalopathies. Applicant: Dr. Ziobro holds M.D. and Ph.D. degrees and has completed specialty clinical training in both Child Neurology and Pediatric Epilepsy. She has previous experience in neuroscience research using rodent, neuronal cell culture, and organotypic hippocampal cell culture models of acquired epilepsy. This career development plan includes a period of mentored research designed to develop the applicant’s knowledge in advanced imaging techniques, viral vector design, brain slice electrophysiology, and interneuron development. These skills will complement her current skill-set and propel her development as an independent researcher. The concepts learned during this training period will be broadly applicable to multiple disease processes and allow for significant clinical translation in a clinically relevant field of study. Research Plan: PCDH19-clustering epilepsy (PCE) is one of the most common monogenic developmental and epileptic encephalopathies (DEEs), characterized by cognitive impairment and intractable seizure clusters starting in infancy. PCDH19 is and X-linked gene that encodes a transmembrane cell adhesion molecule, critical for cell interactions during brain development. PCE affects females and rare mosaic males, while males expressing only mutant PCDH19 do not develop epilepsy. A leading hypothesis to explain this phenomenon is that it occurs due to cellular interference associated with random X-inactivation (or mosaic mutations) in which cells expressing wild type and those expressing mutant PCDH19 fail to interact properly during brain development, which is supported by a preliminary data showing a unique cell segregation pattern of Pcdh19+ and Pcdh19- neurons in the cortex and hippocampus of the PCE mouse model. Our central hypothesis is that mosaic Pcdh19 expression alters interneuron development in the hippocampal CA1 region leading to aberrant network formation, hyperexcitability and increased seizure susceptibility. This proposal will characterize histologic (Aim 1) physiologic (Aim 2) and developmental (Aim 3) mechanisms of PCE. This study will provide significant insight into the mechanisms of PCE and guide potential therapeutic strategies that may be applicable to multiple genetic epilepsies.