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：
9268811
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 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9268811
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 Gray unit of radiation dose Hippocampus (Brain)Hour Injectable Intractable Epilepsy Intravenous Ion Channel Kainic Acid Libraries Mechanics 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 public health relevance 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）：癫痫病在美国约300万人，尽管当前的抗癫痫药有效地控制着大约70％的人群的癫痫发作，但药物不能充分控制其余30％的癫痫发作（Kwan and Brodie，2000年）。由于不到10％的药物难治性癫痫患者被认为是手术切除，因此大量癫痫患者基本没有有效的治疗选择（Engel等人； 1992； Siegel，2004）。即使引入了许多新药，耐药性癫痫的数量尚未减少，这促使Loscher和Schmidt（2011）指出：“可用证据表明癫痫药物治疗的疗效和耐受性并未显着改善”。 Gray等人（2010年）的最新研究为该问题提供了潜在的解决方案。使用急性边缘癫痫发作模型Gray等。（2010年）表明，衣壳DNA改组和定向进化可以鉴定出一种新型的，嵌合腺体相关的病毒（AAV）克隆（＃83），该病毒（＃83）在静脉内施用后选择性地越过了急性癫痫发作，急性癫痫发作损害了CNS中的急性癫痫发作，并损害了CNS中的细胞。为了实现这种方法的全部潜力，我们假设慢性边缘癫痫发作模型中的其他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 CAPSID库，一个基于Clone 83的误差PCR，另一个是由独特的多个克隆组成的，这些克隆越过癫痫发作损害血脑屏障。文库将静脉注射到具有记录的慢性自发癫痫发作活动的大鼠中。随后，神经元将与海马和梨状皮层分离，并将拯救突变克隆。在体内验证转导功效和周围生物分布之后，将生产最有效的克隆，将最有效的克隆包装，并将产生经过验证的治疗性神经肽盒和重组病毒。这些新颖的向量将静脉内给予具有记录的自发癫痫发作活性的大鼠，以评估衰减自发的边缘癫痫发作活性的能力，以及急性癫痫发作后测试抗癫痫发作的能力。如果成功，发现将极大地改变当前的癫痫治疗范例，并显着影响治疗难治性癫痫群。