Mechanisms of PCDH19-clustering epilepsy

项目总结/摘要 该提案描述了一个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，研究机制和潜在的治疗发展和 癫痫性脑病 申请人：Ziobro博士拥有医学博士学位。和博士学位，并已完成专业临床培训，在这两个孩子 神经病学和小儿癫痫。她以前有过利用啮齿动物进行神经科学研究的经验， 神经元细胞培养和获得性癫痫的器官型海马细胞培养模型。这个职业 发展计划包括一段时间的指导研究，旨在发展申请人的知识， 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，并推动她作为一个独立的研究人员的发展。 培训期间学到的概念将广泛适用于多种疾病过程， 允许在临床相关研究领域进行重要的临床翻译。 研究计划：PCDH 19聚集性癫痫（PCE）是最常见的单基因发育性癫痫之一， 和癫痫性脑病（DEE），以认知障碍和顽固性癫痫簇为特征 从婴儿期开始PCDH 19是编码跨膜细胞粘附分子的X连锁基因， 在大脑发育过程中细胞相互作用的关键。PCE影响女性和罕见的马赛克男性，而男性 仅表达突变型PCDH 19的小鼠不发生癫痫。解释这一现象的一个主要假设是 它的发生是由于与随机X失活（或嵌合突变）相关的细胞干扰， 表达野生型的细胞和表达突变型PCDH 19的细胞在脑发育过程中不能正确地相互作用， 这一点得到了初步数据的支持，初步数据显示Pcdh 19+的独特细胞分离模式。 和PCE小鼠模型的皮质和海马中的Pcdh 19-神经元。我们的核心假设是， 嵌合体Pcdh 19表达改变海马CA 1区中间神经元的发育， 网络形成、过度兴奋和癫痫发作易感性增加。该提案将体现 PCE的组织学（目的1）、生理学（目的2）和发育学（目的3）机制。本研究将提供 对PCE的机制有重要的了解，并指导潜在的治疗策略， 适用于多种遗传性癫痫

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.