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发作疾病临床管理的主要障碍，影响三分之一的癫痫患者。此外，遗传性全身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身性癫痫抗癫痫药物反应的细胞和网络基础。候选人在癫痫方面有很强的临床背景，并开展了重要的初步工作，为拟议的研究奠定了基础。该建议的中心假设是，发作间期相对伽马功率（30-100 Hz）可以预测失神癫痫和婴儿期严重肌阵挛癫痫（Dravet综合征）的抗癫痫药物反应，因为这些药物对快速尖峰中间神经元的影响。癫痫的stargazer和摇摇欲坠的小鼠模型，以及Dravet综合征的Scn 1a杂合敲入模型，是研究这一假设的理想模型，因为它们具有与快速尖峰中间神经元缺陷相关的突变，并且这些中间神经元对于新皮质γ节律的产生至关重要。此外，已知这些模型与某些抗癫痫药物具有矛盾的癫痫发作加重。在该项目中，使用体内双光子显微镜和同步EEG、事后免疫组织化学和2 kHz的体内视频EEG监测采样，该项目的具体目标是：（1）在3种遗传性全身性癫痫模型中测定体内抗癫痫药物对新皮层细胞特异性和局部网络的反应，（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2-300 Hz脑电功率的影响。该提案合并了以前在Sydney Cash，MD，PhD的指导下获得的EEG计算分析技术，以及在Jeffrey Noebels，MD，PhD（主要导师）和Stelios Smirnakis，MD，PhD（共同导师）的持续指导下获得的最新技术，前者提供神经发生方面的专业知识，后者提供双光子成像方面的专业知识。在短期内，候选人已经为严格的培训和课程制定了一个基于差距的计划，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转化研究方法的专业知识，同时还学习和治疗癫痫患者的临床背景。从长远来看，凭借德克萨斯州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为独立的临床医生-科学家，其独特的重点是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身性癫痫的机制提供新的线索，并可以直接改善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.