Development of Intravenous AAV Vectors for Intractable Epilepsy

治疗难治性癫痫的静脉 AAV 载体的开发

基本信息

批准号：
9057144
负责人：
Thomas J. McCown
金额：
$ 33.25万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-15 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9057144
关键词：
Acute Address Adverse effects American Animal Model Antiepileptic Agents Antiepileptogenic Area Attenuated Biodistribution Blood - brain barrier anatomy Brain Capsid Caring Cellular Tropism Chronic DNA DNA Shuffling Dependovirus Development Diagnosis Directed Molecular Evolution Dorsal Dose-Limiting Drug resistance Electroencephalography Epilepsy Excision Exhibits Galanin Genes Health Hippocampus (Brain)Hour Intractable Epilepsy Intravenous Ion Channel Kainic Acid Libraries Modeling Mutation Neurons Neuropeptides Operative Surgical Procedures Organ Outcome Outcome Study Patients Peripheral Pharmaceutical Preparations Pharmacotherapy Population Publications Rattus Refractory Research Seizures Signal Pathway Testing Therapeutic Therapeutic Effect Time Translating Treatment Efficacy Validation adeno-associated viral vector base cellular transduction experience gene therapy improved in vivo innovation intravenous administration mutant neutralizing antibody novel novel therapeutic intervention novel therapeutics piriform cortex receptor recombinant virus small molecule vector viral DNA

项目摘要

DESCRIPTION (provided by applicant): Epilepsy afflicts approximately 3 million people in the U.S., and although current, anti-epileptic medication effectively controls the seizures in approximately 70% of this population, medications do not adequately control seizures for the remaining 30% (Kwan and Brodie, 2000). Because fewer than 10% of patients with drug refractory epilepsy are considered for surgical resection, a substantial number of epilepsy patients essentially have no effective therapeutic options (Engel et al.; 1992, Siegel, 2004). Even with the introduction of many new drugs, the number of drug resistant epilepsies has not decreased, prompting Loscher and Schmidt (2011) to state that "the available evidence indicates that the efficacy and tolerability of drug treatment of epilepsy has not substantially improved". Recent studies by Gray et al (2010) have provided a potential solution to this problem. Using an acute limbic seizure model Gray et al. (2010) showed that capsid DNA shuffling and directed evolution could identify a novel, chimeric adeno-associated virus (AAV) clone (#83) which upon intravenous administration selectively crossed the acute seizure compromised blood-brain barrier and transduced cells in the CNS. In order to realize the full potential of this approach we hypothesize that additional DNA shuffling and directed evolution in a chronic limbic seizure model will produce safe, therapeutically effective, chimeric vectors. Two new brain specific AAV capsid libraries will be constructed, one based upon error prone PCR of clone 83 and the other composed of unique, multiple clones that cross the seizure compromised blood-brain barrier. The libraries will be injected intravenously into rats with documented chronic, spontaneous seizure activity. Subsequently, neurons will be dissociated from the hippocampus and the piriform cortex, and the mutant clones will be rescued. After in vivo validation of both transduction efficacy and peripheral biodistribution, the most effective clones will be packaged with proven therapeutic neuropeptide cassettes and recombinant virus will be produced. These novel vectors will be administered intravenously to rats with documented spontaneous seizure activity in order to assess the ability to attenuate spontaneous limbic seizure activity, as well as after acute seizures to test for anti-epileptogenic actions. If successful the findings would dramatically shift current epilepsy treatment paradigms and significantly impact the treatment refractory epileptic population.

描述（由申请人提供）：在美国，癫痫症困扰着大约 300 万人，尽管目前的抗癫痫药物有效控制了该人群中约 70% 的癫痫发作，但药物并不能充分控制其余 30% 的癫痫发作（Kwan和布罗迪，2000）。由于只有不到 10% 的药物难治性癫痫患者被考虑进行手术切除，因此大量癫痫患者基本上没有有效的治疗选择（Engel 等人；1992 年；Siegel，2004 年）。即使推出了许多新药，耐药性癫痫的数量并没有减少，促使Loscher和Schmidt（2011）指出“现有证据表明癫痫药物治疗的疗效和耐受性并没有实质性改善”。 Gray等人（2010）最近的研究为这个问题提供了一个潜在的解决方案。使用急性边缘癫痫模型 Gray 等人。 (2010) 表明衣壳 DNA 改组和定向进化可以识别一种新型嵌合腺相关病毒 (AAV) 克隆 (#83)，该克隆在静脉注射后选择性地穿过急性癫痫发作受损的血脑屏障并转导中枢神经系统细胞。为了充分发挥这种方法的潜力，我们假设慢性边缘癫痫模型中额外的 DNA 改组和定向进化将产生安全、治疗有效的嵌合载体。将构建两个新的大脑特异性 AAV 衣壳文库，一个基于克隆 83 的容易出错的 PCR，另一个由独特的多个克隆组成，这些克隆可以穿越癫痫受损的血脑屏障。这些文库将被静脉注射到有记录的慢性自发性癫痫发作活动的大鼠体内。随后，神经元将与海马体和梨状皮层分离，突变克隆将被拯救。在体内验证转导功效和外周生物分布后，最有效的克隆将与经过验证的治疗性神经肽盒一起包装，并生产重组病毒。这些新型载体将通过静脉注射给予记录有自发性癫痫发作活动的大鼠，以评估减弱自发性边缘系统癫痫发作活动的能力，以及在急性癫痫发作后测试其抗癫痫作用。如果成功，这些发现将极大地改变当前的癫痫治疗模式，并对难治性癫痫人群产生重大影响。