Impaired KCC2 function underlies pharmacoresistant seizures

描述(申请人提供)：癫痫是一种可治愈的疾病，但近四分之一的患者出现对抗癫痫药物产生抗药性的癫痫发作。癫痫是以脑电波异常和抑制减弱为特征的神经系统疾病。大脑抑制的基本决定因素是KCC2蛋白，它决定了关键的抗癫痫药物靶点如何工作。如果没有KCC2，这些靶点将不再正常发挥作用，因此，一线和二线抗癫痫药物疗法也不会。最近才发现KCC2在耐药癫痫患者的大脑中严重减少。这项提议是第一次明确地将这种蛋白的丢失确定为耐药癫痫的共同特征。鉴于这种蛋白质的基本作用，建议的实验必须完成，以造福于患有这种尚未治愈的疾病的人。该项目将包括一系列分子、电生理和遗传学实验，以确认KCC2在耐药癫痫动物模型中的假设作用。第一个项目的目的是利用一种新的策略在分子水平上检测受损的KCC2功能。这将证明KCC2功能的缺陷阻碍了脑细胞之间的抑制性信号传递。第二个项目的目的是利用动物脑组织中耐药癫痫的模型来直接证明KCC2功能缺陷导致了这种疾病。这将是在任何耐药癫痫模型中这一过程的第一次演示。最终的项目目标将直接解决医学中一个长期悬而未决的问题。多年来，医生们已经知道，耐药癫痫会随着时间的推移而发展，从可以治疗的状态缓慢地、有时甚至是快速地演变为无法治疗的状态。这些实验将是在活的动物大脑中首次证明KCC2功能的进行性丧失是这些耐药癫痫发生的基础。这一发现表明，挽救KCC2功能可以恢复现有抗癫痫药物的疗效。主要的研究目标是 为立即测试一种新的治疗策略奠定基础，该策略针对KCC2功能，以改善耐药癫痫患者的生活质量。

DESCRIPTION (provided by applicant): Epilepsy is a curable disease, but nearly one out of four patients develops seizures that are resistant to anti- epileptic medications. Seizures are neurological conditions characterized by abnormal brain waves and decreased inhibition. The fundamental determinant of inhibition in the brain is the protein KCC2, which dictates how key anti-epileptic drug targets work. Without KCC2 these targets would no longer function properly, and consequently, neither would the first- and second-line anti-epileptic drug therapies. KCC2 was only recently found to be severely diminished in the brains of people suffering from drug-resistant epilepsy. This proposal represents the first attempt to unequivocally identify the loss o this protein as the common feature of drug- resistant seizures. Given the fundamental role of this protein, it is vital that the proposed experiments are completed for the benefit of people suffering from this as yet incurable disorder. The project will consist of an array of molecular, electrophysiological, and genetic experiments to confirm the hypothesized role of KCC2 in animal models of drug-resistant seizures. The first project aim utilizes a new strategy to examine impaired KCC2 function on the molecular scale. This will demonstrate that a deficit in KCC2 function impedes inhibitory signaling between brain cells. The second project aim utilizes a model of drug-resistant seizures in animal brain tissue to directly demonstrate that the deficit in KCC2 function causes this disorder. This will be the first demonstration of this process in any model of drug-resistant seizures. The final project aim will directly address a longstanding unanswered question in medicine. Physicians have known for years that drug-resistant seizures develop with time, slowly and sometimes rapidly evolving from a state that can be treated into one that cannot. These experiments will be the first demonstration in a living animal brain that the progressive loss of KCC2 function underlies the development of these drug-resistant seizures. Such a finding would suggest that rescuing KCC2 function could restore the therapeutic effectiveness of current anti-epileptic drugs. The overarching research objective is to lay the foundation for immediate testing of a novel therapeutic strategy that targets KCC2 function to improve the quality of life of those suffering from drug-resistant seizures.