Neural circuit mechanisms controlling seizures

颞叶癫痫（TLE）是成人中最常见的癫痫形式，与显著的认知能力下降相关。在超过40%的TLE病例中，目前的治疗方案无法控制癫痫发作，全身性抗癫痫药物给药可能会产生重大的负面副作用，因此需要更有效的治疗方法。然而，由于研究慢性自发性癫痫发作的固有挑战，TLE的细胞和电路机制尚未被理解，慢性自发性癫痫发作通常发生在相对较短的时间窗口内，通常在几秒到几分钟的时间尺度上。使用最近开发的分子工具，该工具整合了光和钙，以在短时间窗内标记活性细胞，沿着用于癫痫发作检测和光传递的闭环系统，初步结果确定了海马内的一个独特的细胞簇在癫痫发作期间显著活跃。额外的初步工作确定该区域也参与发作间期癫痫样事件，这表明它是癫痫发作发展的关键控制节点。该提案将采用两种不同的TLE模型，即最近开发的局灶性基因敲除模型和海马内钾盐模型，来剖析该系综的作用，并1）确定其参与慢性癫痫动物的发作间隙和发作活动;（二）确定癫痫发作期间这些细胞的光遗传学抑制是否可以使用转基因药物控制慢性自发性癫痫发作及其相关的认知共病。小鼠系，其提供接近该独特细胞群的途径;和3）确定在该区域中的早期干预是否可以预防癫痫及其相关认知共病的进展。候选人已经组建了一个由Gyorgy Buzsaki，Liqun Luo和Alice Ting组成的咨询委员会，以支持获得神经元振荡闭环控制，清除组织成像和基因表达分析以及分子工具开发方面的额外培训。此外，候选人还提出了一个个性化的职业发展计划，包括在赠款写作，教学和科学管理方面的额外经验，以促进独立研究人员的成功。候选人的长期目标是发展作为独立神经科学家的职业生涯，利用分子、细胞、电路和行为水平的多尺度研究来了解神经元功能及其在癫痫等神经系统疾病中的功能障碍的机制。完成拟议的研究将通过以下方式推进该领域：1）建立发作间期和发作活动之间先前未知的关系; 2）确定在海马体先前未探索的区域中干预的治疗潜力，以控制癫痫发作和相关的认知缺陷; 3）确定海马体特异性细胞系综以供进一步研究。K99/R 00奖学金提供的培训期将使候选人能够发展一套强大的技能和资源，用于她的独立职业生涯，斯坦福大学的跨学科性质为候选人成功开展研究和培训计划提供了理想的环境。

Temporal lobe epilepsy (TLE) is the most common form of epilepsy in adults and is associated with significant cognitive decline. In over 40% of TLE cases, seizures are not controlled with current treatment options and systemic anti-epileptic drug administration can have major negative side effects, prompting the need for more effective therapies. However, the cellular and circuit mechanisms underlying TLE are not yet understood due to the inherent challenges of studying chronic spontaneous seizures which typically occur within a relatively short temporal window, often on a timescale of seconds to minutes. Using a recently developed molecular tool, which integrates light and calcium to label active cells within a short temporal window, along with a closed-loop system for seizure detection and light delivery, preliminary results identified a distinct cluster of cells within the hippocampus prominently active during seizures. Additional preliminary work identifies this region as also involved during interictal epileptiform events, suggesting it is a critical control node in the development of seizures. This proposal will employ two different models of TLE, a recently developed focal genetic knockout model and the intrahippocampal kainite model, to dissect the role of this ensemble and 1) Determine its involvement in both interictal and ictal activity in chronically epileptic animals; 2) Determine whether optogenetic inhibition of these cells during seizures can control chronic spontaneous seizures and its associated cognitive comorbidities using a transgenic mouse line that provides access to this distinct population of cells; and 3) Determine whether early intervention in this region can prevent the progression of epilepsy and its associated cognitive comorbidities. The candidate has assembled an Advisory Committee comprised of Gyorgy Buzsaki, Liqun Luo, and Alice Ting to support the acquisition of additional training in closed-loop control of neuronal oscillations, cleared tissue imaging and gene expression analysis, and molecular tool development. In addition, the candidate proposes a personalized plan for career development comprised of additional experience in grant writing, teaching, and scientific management to facilitate success as an independent researcher. The candidate’s long-term goal is to develop a career as an independent neuroscientist utilizing multi-scale investigation at the level of molecules, cells, circuits, and behavior to understand mechanisms of neuronal function and their dysfunction in neurological disorders such as epilepsy. Completion of the proposed study will advance the field by 1) Establishing a previously unknown relationship between interictal and ictal activity; 2) Identifying the therapeutic potential of intervention in a previously unexplored area of the hippocampus to control seizures and associated cognitive deficits; and 3) Identify seizure-specific cellular ensembles for further study. The training period afforded by the K99/R00 award will allow the candidate to develop a powerful set of skills and resources to use in her independent career and the interdisciplinary nature at Stanford provides the ideal environment for the candidate to carry out the research and training plan successfully.