Neural circuit mechanisms controlling seizures

颞叶癫痫(TLE)是成人最常见的癫痫类型，与认知功能显著下降有关。在超过40%的TLE病例中，目前的治疗方案无法控制癫痫发作，全身抗癫痫药物的使用可能会产生重大的负面副作用，促使人们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内在挑战，TLE的细胞和电路机制尚不清楚。慢性自发性癫痫发作通常发生在相对较短的时间窗口内，通常是在几秒到几分钟的时间尺度上。利用最近开发的分子工具，将光和钙整合在一起，在短时间窗口内标记活动细胞，以及用于癫痫检测和光传递的闭环系统，初步结果发现，在癫痫发作期间，海马体内有一群明显活跃的细胞。更多的初步工作发现，这个区域也参与了发作间期的癫痫样事件，这表明它是癫痫发作发展的关键控制节点。这项建议将使用两种不同的TLE模型，即最近开发的局部基因敲除模型和海马内Kainite模型，以剖析这一整体的作用，并1)确定它是否参与慢性癫痫动物的发作间期和发作活动；2)确定在癫痫发作期间对这些细胞的光遗传抑制是否可以控制慢性自发性癫痫发作及其相关的认知并发症；以及3)确定该区域的早期干预是否可以阻止癫痫的进展及其相关的认知共生问题。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，以支持获得有关神经元振荡的闭环控制、透明组织成像和基因表达分析以及分子工具开发的额外培训。此外，候选人还提出了一项个性化的职业发展计划，其中包括额外的拨款撰写、教学和科学管理方面的经验，以促进作为独立研究人员的成功。候选人的长期目标是发展成为一名独立的神经科学家，利用分子、细胞、电路和行为水平的多尺度研究，了解癫痫等神经疾病中神经功能及其功能障碍的机制。这项拟议研究的完成将推动这一领域的发展，1)建立发作间期和发作活动之间以前未知的关系；2)确定在以前未探索的海马区进行干预以控制癫痫发作和相关认知缺陷的治疗潜力；以及3)确定癫痫发作特有的细胞群，以供进一步研究。K99/R00奖项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实施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.