An Interneuron-based Cell Therapy for Epilepsy

描述（由申请人提供）：神经祖细胞移植对神经系统疾病的治疗提出了很大的期望。最近的一些研究，包括我们实验室的研究（Wichertle et al. 1999; Alvarez-Dolado et al. 2006）表明，来自胚胎内侧神经节嵴（MGE）的移植祖细胞具有独特的分散、迁移和分化为gaba能中间神经元的能力。虽然我们最近证明这些细胞在癫痫突变小鼠模型中有效抑制自发发作（Baraban et al. 2009），但这种方法的治疗潜力在很大程度上仍未被探索。针对最近NINDS癫痫研究的“基准”，我们在此提出了利用小鼠胚胎或小鼠胚胎干细胞衍生的MGE祖细胞开发成人移植策略的实验。这些细胞将在自发发作后移植到颞叶癫痫的普通啮齿动物模型中。技术将包括使用体外保存的急性脑切片，以及应用可视化膜片钳方法来研究mge衍生的中间神经元的功能整合。视频脑电图监测和免疫荧光技术也将应用。提出了三个具体目标：(1)开发、优化和验证成年小鼠海马移植MGE祖细胞的方法；(2)研究成年MGE祖细胞移植在小鼠颞叶癫痫模型中的治疗潜力；(3)从小鼠胚胎干细胞中开发MGE祖细胞系。我们的结果有望为移植的MGE祖细胞在成人宿主脑中的功能提供新的信息，并可能直接证明祖细胞治疗癫痫的潜力。

DESCRIPTION (provided by applicant): Transplantation of neural progenitor cells raises great expectations for the treatment of neurological disorders. A number of recent studies, including those from our laboratory (Wichertle et al. 1999; Alvarez-Dolado et al. 2006), demonstrate that transplanted progenitor cells derived from the embryonic medial ganglionic eminence (MGE) possess a unique ability to disperse, migrate, and differentiate into GABAergic interneurons. Although we recently demonstrated that these cells are effective in suppressing spontaneous seizures in a mutant mouse model of epilepsy (Baraban et al. 2009), the therapeutic potential of this approach remains largely unexplored. In response to recent NINDS epilepsy research "benchmarks", we here propose experiments to develop an adult transplantation strategy using MGE progenitors harvested from mouse embryos or derived from mouse embryonic stem cells. These cells will be transplanted in a common rodent model of temporal lobe epilepsy after the emergence of spontaneous seizures. Techniques will involve use of acute brain slices maintained in vitro, and application of visualized patch clamp methods to study the functional integration of MGE-derived interneurons. Video-EEG monitoring and immunofluorescence techniques will also be applied. Three specific aims are proposed: (i) to develop, optimize, and validate a method for transplantation of MGE progenitor cells in the adult mouse hippocampus (ii) to examine the therapeutic potential of adult MGE progenitor cell grafts in a mouse model of temporal lobe epilepsy, and (iii) to develop an MGE progenitor cell line from mouse embryonic stem cells. Our results promise to provide new information about the function of transplanted MGE progenitors in the adult host brain and may provide a direct demonstration of the potential for progenitor cells to treat epilepsy. PUBLIC HEALTH RELEVANCE: Epilepsy is a common neurological disorder afflicting nearly 3 million Americans. Given that loss or reduction of inhibitory synaptic transmission in hippocampus is one potential mechanism resulting in the emergence of epilepsy, a method to generate new hippocampal interneurons could have direct therapeutic consequences. Using mouse embryonic and stem cell derived neural progenitor cells and a common rodent model of temporal lobe epilepsy we propose to develop a novel interneuron-based cell therapy.