EnCRISPrx - A Permanent CRISPR-based Approach To Rescue Dravet Syndrome By Enhancing Scn1a Promoter Activity

Dravet综合征是一种影响幼儿的严重灾难性神经系统疾病。全世界每天都有3到4名儿童被诊断出患有Dravet综合征，症状包括癫痫、自闭症、运动障碍和睡眠障碍。迄今为止，大多数治疗无效或耐受性差。钠离子通道药物对多种形式的癫痫有效，但也可能使Dravet综合征恶化。在大约80%的病例中，Dravet综合征是由单个基因SCN 1A中的数百个杂合突变之一引起的，这些突变导致功能丧失。这些是自发的或遗传的，并导致蛋白质表达不足。我们的目标是通过增加蛋白质表达的单一治疗来挽救所有这些突变。为此，我们建议修改SCN 1A基因，使其更具活性。我们将增加基因的两个拷贝的活性，但只有非突变的一个会产生新的蛋白质。我们的目标是一种能够恢复大脑正常功能的疗法，这与传统药物不同，传统药物的目标是大脑中沟通的基础蛋白质，并且可以抑制整体活动。我们将在Dravet综合征的动物模型中测试几种可能的方法，使用神经元兴奋性，癫痫和行为变化作为读数。这一建议不仅有助于了解Dravet综合征的一般治疗是否可行，而且还将深入了解这些技术治疗由不同基因中类似突变引起的其他神经系统疾病的潜力。我们强调，这里提出的方法不是基因编辑本身，它一次只能拯救一个突变。相反，我们的目标是纠正导致共同病理的所有突变的影响，因此该策略可以推广，并对具有共同遗传机制的广泛疾病产生重大影响。

Dravet Syndrome is a severe catastrophic neurological disorder affecting young children. Every day 3 or 4 children are diagnosed with Dravet syndrome worldwide, with symptoms including epilepsy, autism, movement disorders and sleep disturbances. To date, the majority of therapies are ineffective or poorly tolerated. Sodium channel drugs which are otherwise effective in many forms of epilepsy may even worsen Dravet syndrome.In approximately 80% of cases, Dravet syndrome is caused by one of several hundred heterozygous mutations in a single gene, SCN1A, which result in loss of function. These are either spontaneous or inherited, and lead to insufficient expression of the protein. Our aim is to rescue all of these mutations with a single treatment that increases the expression of the protein. To do so, we propose to modify SCN1A gene to make it more active. We will increase the activity of both copies of the gene, but only the non-mutant one will make new proteins. We are aiming for a therapy that will restore the normal functioning of the brain, which is unlike conventional drugs that target proteins that underlie communication in the brain and can dampen activity overall. We will test several possible approaches in an animal model of Dravet Syndrome, using neuronal excitability, epilepsy and behavioural changes as readouts. This proposal will be useful not only for understand if a general treatment for Dravet syndrome is achievable, but also will give insight into the potential of these techniques to cure other neurological diseases caused by similar mutations in different genes. We stress that the approach proposed here is not gene editing as such, which can rescue only a single mutation at a time. Instead, we aim to correct the effects of all the mutations leading to a common pathology, and so the strategy can be generalized and have a substantial impact for a wide range of diseases that share a common genetic mechanism.