Blood-brain barrier gene delivery in knock-out mice.

描述（由申请人提供）：除了Lafora进行性肌阵挛性癫痫外，还有许多单基因缺陷疾病对成年携带者的子女具有破坏性影响（例如Rett综合征、脆性X综合征、Canavan病和Tay-Sachs病）。对于所有这些疾病，都有由患病儿童的父母和朋友组成的支持小组。在所有情况下，突变基因都是已知的并被克隆。但是，由于在整个大脑中表达外源基因的问题，这些潜在的救命基因的副本在研究实验室中处于休眠状态。由于难以穿过脑毛细血管递送大分子治疗剂，因此认为血脑屏障（BBB）可能是一个无法解决的问题。英雄的措施，如短暂中断血脑屏障与渗透压休克，尚未进行评估的基因治疗药物在临床试验中交付。替代方法也不是没有问题;在今年，NIH停止了所有使用病毒载体进行基因治疗的试验。这一行动表明，需要开发和测试非病毒替代品，以通过BBB传递（大分子）基因。 我们将测试的假设，最近开发的免疫脂质体血脑屏障传递系统可用于成功地促进基因治疗，通过血脑屏障敲除小鼠与Lafora的进行性肌阵挛癫痫（PME），静脉内给药后。我们建议，如果这种非病毒递送系统可以在动物模型中治疗这种疾病，则可以开发用于临床使用的基于免疫脂质体的治疗这种致命癫痫的方法。其目标是：（1）制备用于递送正常EPM 2a/拉福林基因的聚乙二醇化免疫脂质体（PIL）;（2）在拉福拉病的敲除小鼠模型中施用PIL;（3）确认静脉内注射后基因均匀递送至脑，并改善进行性疾病;和（4）开发一种最佳的治疗方案，其在基因敲除小鼠中阻止Lafora病的致命发作。

DESCRIPTION (provided by applicant): In addition to Lafora's Progressive Myoclonic Epilepsy, there are numerous single-gene defect diseases that have devastating effects on the children of adult carriers (e.g. Rett's syndrome, fragile x syndrome, Canavan's disease, and Tay-Sachs disease). For all of these diseases, there are support groups formed by parents and friends of afflicted children. In all cases, the mutated gene is known and cloned. But copies of the potentially life-saving genes sit dormant in research laboratories because of problems in expressing an exogenous gene throughout the brain. Due to the difficulties in delivering large molecule therapeutics across the brain capillaries, there is a perception that the blood-brain barrier (BBB) may be an insoluble problem. Heroic measures, such the transient disruption of the BBB with osmotic shock, have yet to be evaluated for delivery of gene therapeutics in clinical trials. Alternative methods are also not without problems; within the current year the NIH placed a halt on all trials where viral vectors were being used to deliver gene therapies. This action suggests a need to develop and test non-viral alternatives for the delivery of (large-molecule) genes across the BBB. We will test the hypothesis that recently developed immunoliposome BBB delivery systems can be used to successfully promote a gene therapeutic through the BBB of knock-out mice with Lafora's Progressive Myoclonic Epilepsy (PME), after intravenous administration. We propose that if this non-viral delivery system can treat the disease in animal models, an immunoliposome-based cure for this fatal epilepsy can be developed for clinical use. The aims are: (1) To prepare pegylated immunoliposomes (PIL) for delivery of the normal EPM2a/laforin gene; (2) to administer PILs in a knockout mouse model of Lafora's Disease; (3) to confirm uniform delivery of the gene to the brain after intravenous injection, with amelioration of the progressive disease; and (4) to develop an optimal therapeutic regimen which arrests the fatal onset of Lafora's disease in the knock-out mice.