Developing a drug-inducible gene therapy for temporal lobe epilepsy

开发药物诱导的颞叶癫痫基因疗法

基本信息

批准号：
10800000
负责人：
EDWARD PEREZ-REYES
金额：
$ 53.15万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-07-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10800000
关键词：
Address Affect Agonist American Animal Model Anticonvulsants Basic Science Brain region Cells Characteristics Clinical Collaborations Convulsants Development Disease Dose Doxycycline Drug Screening Electric Stimulation Elements Enhancers Environment Epilepsy Equilibrium FDA approved Frequencies Gene Expression Goals Grant Hippocampus Hybrids Immune Kainic Acid Learning Maps Medical Medicine Memory Modeling Motor Seizures Mouse Strains Mus Neuromodulator Neurons Partial Epilepsies Patients Persons Pharmaceutical Preparations Pharmacotherapy Potassium Channel Randomized Recurrence Reproducibility Research Research Design Rodent Model Safety Seizures Side Site Sleep Structure System Technology Temporal Lobe Epilepsy Testing Universities Viral Vector antagonist clinical translation design drug development efficacy study excitatory neuron forest gamma-Aminobutyric Acid gene product gene therapy immunocytochemistry inhibitory neuron insight miniaturize neural circuit neuronal circuitry next generation novel overexpression preclinical study promoter side effect success targeted treatment

项目摘要

There are 3 million Americans with epilepsy. This research aims to develop a first-in-class drug- inducible gene therapy for the most common form of focal epilepsy, temporal lobe epilepsy (TLE). TLE affects half a million Americans, yet despite the size of this problem, medical treatment of TLE fails in a third of these patients. Clearly there is a large unmet clinical need for a treatment without side effects. The guiding hypothesis of these studies is that TLE is a circuit-based disease, requiring the development of next-generation gene therapies that target these circuits. Key technological advances made to address this hypothesis include: 1) development of novel animal models of TLE with spontaneous seizure characteristics that are amenable to drug screening; 2) mapping epileptic circuits using activity-dependent promoters; and 3) development of drug-inducible gene therapies that target specific circuits. It was discovered that electrical stimulation of a specific strain of mouse (VGAT-Cre) was sufficient to trigger limbic epilepsy. To extend the model to other strains of mice, electrical stimulation was combined with a chemoconvulsant, kainic acid. This hybrid approach effectively triggered spontaneous seizures in commonly used mouse strains, such as C57Bl/6. Importantly, the hybrid approach opened the door for studies using the TRAP2 strain, which has proven useful for circuit mapping studies. The drug-inducible gene therapy uses the well-established doxycycline (Dox) regulated system. The technological challenges were to miniaturize this system so it fits in viral vectors, reduce gene expression in the absence of the inducer (leak), and develop a toolkit of promoters to target neuronal subtypes. The goal is to develop a Dox-On system where Dox administration reduces seizures and that possible side effects can be reduced by lowering the Dox dose. Reducing leak to background levels is an important safety feature that has yet to be incorporated in any FDA-approved gene therapy. The mechanisms of action of many current antiseizure drugs are to either reduce the activity of excitatory neurons or enhance the activity of inhibitory neurons. By developing gene therapies that selectively target these types of neurons, the proposed studies will compare these two approaches. Key metrics for these therapies are to show they reduce spontaneous seizure frequency by 50% and target specific nodes in the epileptic circuit. Efficacy studies will use rigorous preclinical study designs with blinding, reproducibility, and target engagement. Contributing to the success of this study is a rich environment for clinical translation of basic research in animal models of epilepsy.

有300万美国人患有癫痫病。这项研究旨在开发一流的药物 - 最常见形式的局灶性癫痫，颞叶癫痫（TLE）的诱导基因治疗。 TLE 影响了50万美国人，但是尽管有这个问题的大小，但TLE的医疗治疗失败了这些患者的三分之一。显然，对没有副作用的治疗有很大的未满足临床需求。这些研究的指导假设是TLE是一种基于电路的疾病，需要开发针对这些电路的下一代基因疗法。关键技术进步解决此假设的解决包括：1）与自发癫痫发作特征，可容纳药物筛查； 2）映射癫痫电路使用活动依赖性启动子； 3）开发靶向靶向的药物诱导基因疗法特定电路。发现特定小鼠菌株的电刺激（VGAT-CRE）足以触发边缘癫痫。为了将模型扩展到其他小鼠菌株，电气将刺激与化学弹药抗体合并。这种混合方法有效在常用的小鼠菌株中触发自发癫痫发作，例如C57BL/6。重要的是，混合方法为使用TRAP2菌株打开了研究的大门，该菌株已被证明对电路有用映射研究。药物诱导的基因疗法使用良好的强力霉素（DOX）受管制系统。技术挑战是使该系统微型化，因此它适合病毒向量，在没有诱导剂（泄漏）的情况下降低基因表达，并将启动子的工具包开发到靶神经元亚型。目的是开发一个DOX-ON系统，其中DOX管理会减少癫痫发作和可能的副作用可以通过降低DOX剂量来减少。减少泄漏背景级别是一项重要的安全功能，尚未纳入任何FDA批准基因疗法。许多当前毒药的作用机制是减少兴奋性神经元的活性或增强抑制性神经元的活性。通过开发基因选择性靶向这些类型神经元的疗法，拟议的研究将比较这两种方法。这些疗法的关键指标是表明它们降低了自发癫痫发作频率在癫痫电路中增加50％和靶向特定节点。功效研究将使用严格的临床前通过盲目，可重复性和目标参与研究设计。为此取得成功做出贡献研究是癫痫动物模型基础研究的临床翻译的丰富环境。