Impaired KCC2 function underlies pharmacoresistant seizures

描述（由申请人提供）：癫痫是一种可治愈的疾病，但近四分之一的患者会出现癫痫发作，并且抗癫痫药物具有耐药性。癫痫是一种神经系统疾病，其特征是脑电波异常和抑制能力下降。大脑中抑制的基本决定因素是蛋白质KCC2，它决定了关键的抗癫痫药物靶点如何起作用。如果没有KCC2，这些靶点就不能正常发挥作用，因此一线和二线抗癫痫药物治疗也不能正常发挥作用。直到最近才发现，KCC2在患有耐药性癫痫的人的大脑中严重减少。这一建议首次尝试明确地将这种蛋白的缺失确定为耐药癫痫发作的共同特征。鉴于这种蛋白质的基本作用，完成拟议的实验对患有这种尚未治愈的疾病的人来说是至关重要的。该项目将包括一系列分子、电生理和基因实验，以确认KCC2在耐药癫痫动物模型中的假设作用。第一个项目目标是利用一种新的策略在分子尺度上检查受损的KCC2功能。这将证明KCC2功能的缺陷会阻碍脑细胞之间的抑制性信号传导。第二个项目的目标是利用动物脑组织中的耐药癫痫模型来直接证明KCC2功能缺陷导致这种疾病。这将是这一过程首次在任何耐药性癫痫发作模型中得到证实。最终的项目目标将直接解决医学中一个长期未解决的问题。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，缓慢而有时迅速地从一种可以治疗的状态演变为一种无法治疗的状态。这些实验将首次在活体动物大脑中证明KCC2功能的逐渐丧失是这些耐药癫痫发作的基础。这一发现提示挽救KCC2功能可以恢复当前抗癫痫药物的治疗效果。首要的研究目标是

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.