Cellular and Network Basis of Anti-Epileptic Drug Response

 描述(由申请人提供)：抗癫痫药物耐药性是癫痫障碍临床治疗的主要障碍，影响三分之一的癫痫患者。此外，遗传性全面性癫痫患者没有癫痫手术的选择。本项目的重点是探索遗传性全面性癫痫抗癫痫药物反应的细胞和网络基础。该候选人在癫痫方面有很强的临床背景，并开展了重要的前期工作，为拟议的研究奠定了基础。这一建议的中心假设是发作间期相对伽马功率(30-100赫兹)可以预测失神癫痫和婴儿期严重肌阵挛癫痫(德拉韦综合征)的抗癫痫药物反应，这是由于这些药物对快速放电的中间神经元的影响。癫痫的观星者和摇摇欲坠的小鼠模型，以及Dravet综合征的Scn1a杂合子敲入模型，都是研究这一假说的理想模型，因为它们的突变与快速放电的中间神经元缺陷有关，而这些中间神经元对新皮质伽马节律的产生至关重要。此外，已知这些模型使用某些抗癫痫药物会出现自相矛盾的癫痫发作加剧。在本项目中，利用体内双光子显微镜和同步脑电、组织后免疫组织化学和2 kHz的在体视频脑电监测采样，本项目的具体目的是：(1)在3种遗传性全身性癫痫模型中，确定体内新皮质细胞特异性和局部网络对抗癫痫药物的反应；(2)在相同的3种遗传性全身性癫痫模型中，评价抗癫痫药物对发作间期脑电功率的影响。这项建议融合了以前在悉尼·卡什医学博士指导下获得的脑电计算分析技术，以及在提供神经发生专业知识的杰弗里·诺贝尔斯医学博士(主要导师)和提供双光子成像专业知识的斯特利奥斯·斯米尔纳基斯博士(共同导师)持续指导下获得的最新技术。在短期内，候选人已经制定了一个基于GAP的计划，用于严格的培训和课程工作，重点是发展体内双光子成像和膜片钳技术、统计建模和分析以及翻译研究方法方面的专业知识，同时还在临床背景下了解和治疗癫痫患者。从长远来看，凭借得克萨斯医学中心强大的机构承诺和丰富的资源，这项培训将有助于候选人发展成为一名独立的临床医生和科学家，专注于减少癫痫患者的耐药性。最终，这项研究的完成将为遗传性全面性癫痫的机制提供新的线索，并可以直接导致改进药物开发和患者护理。

 DESCRIPTION (provided by applicant): Anti-epileptic drug resistance is a major obstacle to the clinical management of seizure disorders and affects one third of patients with epilepsy. In addition, patients with genetic generalized epilepsy do not have the option of epilepsy surgery. The focus of this project is to explore the cellular and network basis of anti-epileptic drug response in genetic generalized epilepsy. The candidate has a strong clinical background in epilepsy and has developed significant preliminary work which forms the basis for the proposed research. The central hypothesis of this proposal is that interictal relative gamma power (30-100 Hz) may predict anti-epileptic drug response in absence epilepsy and severe myoclonic epilepsy of infancy (Dravet Syndrome) due to the effect of these drugs on fast-spiking interneurons. The stargazer and tottering mouse models of epilepsy, as well as the Scn1a heterozygous knock-in model of Dravet Syndrome, are ideal for studying this hypothesis since they have mutations which have been associated with fast-spiking interneuron deficits, and these interneurons are critical for the generation of neocortical gamma rhythms. In addition, these models are known to have paradoxical seizure exacerbation with certain anti-epileptic drugs. In this project, using in vivo 2-photon microscopy and simultaneous EEG, post-hoc immunohistochemistry, and in vivo video-EEG monitoring sampling at 2 kHz, the specific aims of this project are to: (1) Determine the neocortical cell-specific and local network responses to anti-epileptic drugs in vivo in 3 models of genetic generalized epilepsy, and (2) Evaluate the effect of anti-epileptic drugs on interictal EEG power between 2-300 Hz in vivo in the same 3 models of genetic generalized epilepsy. This proposal merges the techniques of computational analysis of EEG previously acquired under the mentorship of Sydney Cash, MD, PhD and more recent techniques acquired under the ongoing mentorship of Jeffrey Noebels, MD, PhD (primary mentor), who provides expertise in neurogenesis, and Stelios Smirnakis, MD, PhD (co-mentor), who provides expertise in 2-photon imaging. In the short term, the candidate has assembled a gap-based plan for rigorous training and coursework focusing on developing expertise in 2-photon imaging and patch-clamp techniques in vivo, statistical modelling and analysis, and translational research methodology, while also learning about and treating patients with epilepsy in a clinical context. In the long term, with a strong institutional commitment and abundant resources available in the Texas Medical Center, this training will aid in the candidate's development as an independent clinician- scientist with a unique focus on diminishing pharmacoresistance in patients with epilepsy. Ultimately, the completion of this research will shed new light on the mechanisms of genetic generalized epilepsy and can directly lead to improved drug development and patient care.