An Interneuron-based Cell Therapy for Epilepsy

描述（由申请人提供）：神经祖细胞的移植为神经系统疾病的治疗带来了很大的期望。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），证明了来自胚胎内侧神经节隆起（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。尽管我们最近证明这些细胞在突变型癫痫小鼠模型中有效抑制自发性癫痫发作（Baraban et al. 2009），但这种方法的治疗潜力在很大程度上尚未探索。为了响应最近的NINDS癫痫研究“基准”，我们在这里提出了实验，以开发一个成人移植策略，使用MGE祖细胞收获小鼠胚胎或来自小鼠胚胎干细胞。将这些细胞移植到常见的啮齿类动物颞叶癫痫模型中，出现自发性癫痫发作后。技术将涉及使用急性脑切片保持在体外，并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。还将应用视频脑电图监测和免疫荧光技术。提出了三个具体的目标：（i）开发，优化和验证MGE祖细胞在成年小鼠海马中移植的方法（ii）检查成年MGE祖细胞移植物在颞叶癫痫小鼠模型中的治疗潜力，以及（iii）从小鼠胚胎干细胞开发MGE祖细胞系。我们的研究结果有望提供有关移植的MGE祖细胞在成人宿主脑中的功能的新信息，并可能直接证明祖细胞治疗癫痫的潜力。 公共卫生相关性：癫痫是一种常见的神经系统疾病，困扰着近300万美国人。鉴于海马中抑制性突触传递的丧失或减少是导致癫痫出现的一种潜在机制，产生新的海马中间神经元的方法可能具有直接的治疗效果。利用小鼠胚胎和干细胞衍生的神经前体细胞和常见的啮齿动物颞叶癫痫模型，我们提出开发一种新的基于中间神经元的细胞治疗。

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.