An Interneuron-based Cell Therapy for Epilepsy

项目摘要/摘要 细胞移植，使用整合在宿主大脑回路中的GABA能中间神经元，其功能类似于 内源性抑制神经元可能为难治性癫痫的治疗提供新的选择。我们的工作， 在过去的二十年中，证明GABA能祖细胞起源于小鼠胚胎内侧 神经节隆起(MGE)整合到宿主回路中(在新生或成年小鼠移植后)， 在那里它们制造功能性抑制性突触，有效地抑制癫痫发作的活动，并纠正 与癫痫相关的行为合并症。建立MGE祖细胞的替代来源 这将有助于将这种治疗疾病的移植策略转化为更大的物种，我们最近 建立了使用猪胚胎MGE祖细胞的方案。在这些发现的基础上，有两个具体的 提出的目标是：(1)优化猪胚胎MGE前体细胞的海马区内异种移植 评估猪胚胎MGE祖细胞可能产生的治疗效果的范围。 技术将涉及尖端电生理学、光遗传和钙成像方法(体外和 体内)研究MGE来源的中间神经元的功能整合。视频脑电监测，啮齿动物癫痫 模型、复杂的行为分析和共聚焦显微镜技术也将被应用。我们的结果 承诺提供有关猪MGE祖细胞的新信息并直接演示其潜力(S) 用于异种移植治疗顽固性癫痫。

Project Summary/Abstract Cell transplantation, using GABAergic interneurons that integrate in host brain circuits, and function similar to endogenous inhibitory neurons could offer a novel therapeutic alternative for intractable epilepsies. Our work, over the past two decades, demonstrated that GABAergic progenitor cells derived from murine embryonic medial ganglionic eminence (MGE) integrate into host circuits (following transplantation in neonatal or adult mice), where they make functional inhibitory synapses, efficiently suppress epileptic seizure activity, and correct behavioral comorbidities associated with epilepsy. To establish an alternative source of MGE progenitor cells that would facilitate translation of this disease-modifying transplantation strategy to larger species, we recently established protocols using porcine embryonic MGE progenitor cells. Building on these findings, two specific aims are proposed: (i) to optimize intra-hippocampal xenotransplantation of porcine embryonic MGE progenitor cells and (ii) to evaluate the range of therapeutic benefits possible with porcine embryonic MGE progenitor cells. Techniques will involve cutting-edge electrophysiology, optogenetic and calcium imaging methods (in vitro and in vivo) to study functional integration of MGE-derived interneurons. Video-EEG monitoring, rodent epilepsy models, sophisticated behavioral assays, and confocal microscopy techniques will also be applied. Our results promise to provide new information about porcine MGE progenitors and direct demonstration(s) of the potential for xenotransplantation as a treatment for intractable epilepsies.