Na Channel beta4 as a Gene Therapy Target for Epilepsy

描述（由申请人提供）：局灶性癫痫发作，如颞叶癫痫（TLE），是一组影响全球数百万人的衰弱性疾病。尽管这个问题很大，但在大多数情况下，对TLE的医疗治疗都失败了。40多年来，这些病人唯一的选择就是手术切除颞叶。因此，开发能够预防局灶性癫痫发作的新疗法将对许多癫痫患者的生活质量产生重大影响。指导R21拨款的假设是，沉默内嗅皮层（EC）中过度活跃的神经元将预防癫痫发作。动物模型和人类患者的大量证据表明，EC神经元在TLE发作中的作用。初步数据显示，在鼠类TLE模型中，EC神经元的放电增加是由于去极化后电流增加和Na通道亚基表达增加。假设是使用短发夹RNA （shRNA）抑制Na通道β -4亚基的表达将减少癫痫发作。重组腺相关病毒（rAAV）已成为人类基因治疗的首选。一个重要的安全特性是调控基因治疗的能力。这将使用新开发的四环素调节盒来完成，该盒控制shRNA的表达并需要药物激活（多西环素- on）。总体目标是开发一种可用于临床的shRNA递送系统，并在TLE动物模型中测试其功效。

DESCRIPTION (provided by applicant): Seizures with a focal onset, such as temporal lobe epilepsy (TLE), are a set of debilitating disorders that affect millions of people worldwide. Despite the size of this problem, medical treatment of TLE fails in most cases. For over 40 years, the only option left for these patients has been surgery to remove the temporal lobe. Therefore, the development of novel treatments that can prevent focal seizures will have a significant impact on the quality of life of many epilepsy patients. The hypothesis guiding this R21 grant is that silencing hyperactive neurons in the Entorhinal Cortex (EC) will prevent seizures. Substantial evidence in both animal models and human patients incriminate the role of EC neurons in TLE seizures. Preliminary data show that in rodent models of TLE, EC neurons have increased firing due to increased after-depolarizing currents and increased expression of Na channel subunits. The hypothesis is that knocking down the expression of the Na channel beta-4 subunit using short-hairpinned RNA (shRNA) will reduce seizures. Recombinant adeno-associated virus (rAAV) has emerged as the top choice for human gene therapy. An important safety feature is the ability to regulate gene therapy. This will be accomplished using a newly developed tetracycline regulator cassette that controls expression of the shRNA and requires drug for activation (doxycycline-ON). The overall goal is to develop a delivery system for the shRNA that can be used in the clinic and to test its efficacy in animal models of TLE.