Innovative systemic gene therapy for treating Parkinson's disease

项目摘要 帕金森病（PD）是一种日益普遍的神经系统疾病，目前约有1 美国有1000万人，全世界有1000万人。尽管最近的创新，大多数先进的 药理学和手术治疗方案最多保持适度的姑息和对症治疗。 基因治疗已经成为一种替代的有希望的手段，以阻止疾病的进展或潜在的 治愈疾病然而，到目前为止，PD基因治疗的临床试验未能建立一个有效的治疗方法。 由于不能实现广泛和有效的基因转移到 大脑中的疾病区域。正在进行的人体试验突出了这个问题的重要性， 其中提高转染的穿透性和效率是主要目标。此外，缺乏可靠的 方法部署基因治疗从血液到脑组织，所有的临床研究，到目前为止， 采用高度侵入性的给药方式，包括将治疗直接注射到脑中。这 事实上，临床试验中排除了早期PD患者，这些患者更有可能对 治疗因此，克服长期存在的系统性基因递送障碍的新方法在整个基因组中是可行的。 PD相关的脑区，包括紧密封闭的血脑屏障（BBB）和密集的神经网络。 脑细胞外基质（ECM）是非常需要的。为此，我们提出了创新的交付方法， 利用：（i）临床上可操作的MR图像引导聚焦超声（FUS）瞬时打开BBB， 基因治疗以靶向方式渗透到脑组织和细胞中，（ii）DNA负载的 纳米颗粒具有独特的能力，可以有效地通过大脑ECM扩散， - 脑内疾病区域中的细胞（即DNA-负载的脑穿透纳米颗粒或DNA-BPN），和 (iii)FUS介导的预处理通过以下方式进一步增强DNA-BPN在脑内的分散： 暂时降低ECM电阻。我们最近在我们的初步研究中表明，FUS介导的靶向BBB 我们的第一代DNA-BPN的渗透和随后的广泛基因转移导致了 常规的基于神经毒素的PD临床前模型的治疗相关基因疗法。作为下一个 迈向临床转化，我们在此建议进一步完善和评估我们的联合交付策略 在家族性和散发性PD的高度复杂和临床相关的临床前模型中， 在人类PD中观察到的病理生理学特征和疾病表型。如果成功，建议 这种方法可以迅速转化为临床使用的基因编码的神经营养因子（即目前 正在进行临床研究，并将在此进行研究），同时可以进行其他临床前研究， 新型遗传靶点在这些高级PD模型中的有效性。此外，该方法还可以 可应用于以脑内高度扩散的疾病区域为特征的其他神经障碍。

PROJECT SUMMARY Parkinson's disease (PD) is an increasingly prevalent neurological disorder that currently affects about one million people in the United States and 10 million worldwide. Despite recent innovations, most advanced pharmacological and surgical therapeutic regimens remain moderately palliative and symptomatic at best. Gene therapy has emerged as an alternative promising means to halt the disease progression or potentially cure the disease. However, clinical trials of PD gene therapy up to this moment have failed to establish a meaningful therapeutic benefit due to an inability to achieve widespread and efficient gene transfer to the disease areas within the brain. The significance of this problem is highlighted by an ongoing human trial, wherein improving the penetration and efficiency of transfection is a primary goal. Further, lacking a reliable method to deploy gene therapy from the bloodstream to the brain tissue, all clinical studies to date have employed highly invasive administration modalities involving direct injection of the therapy into the brain. This reality has precluded the inclusion in clinical trials of early stage PD patients who are more likely to respond to the therapy. Thus, new methods to overcome long-standing barriers to systemic gene delivery throughout the PD-associated brain regions, including the tightly sealed blood-brain barrier (BBB) and the dense network of brain extracellular matrix (ECM), are sorely needed. To this end, we propose innovative delivery approaches exploiting: (i) clinically operable MR image-guided focused ultrasound (FUS) to transiently open the BBB for the penetration of gene therapy into the brain tissues and cells in a targeted manner, (ii) DNA-loaded nanoparticles possessing a unique capability to efficiently spread through the brain ECM to reach and transfect cells in the disease areas within the brain (i.e. DNA-loaded brain-penetrating nanoparticle or DNA-BPN), and (iii) FUS-mediated pre-conditioning that further enhances the dispersion of DNA-BPN within the brain by temporarily reducing ECM resistance. We recently showed in our pilot study that FUS-mediated, targeted BBB penetration of, and subsequent widespread gene transfer by, our first-generation DNA-BPN resulted in therapeutically relevant gene therapy of a conventional neurotoxin-based preclinical model of PD. As a next step towards clinical translation, we here propose to further refine and evaluate our combined delivery strategy in highly sophisticated and clinically-relevant preclinical models of familial and sporadic PD that closely mimic pathophysiological features and disease phenotypes observed in human PD. If successful, the proposed approach could be rapidly translated to the clinic using a gene-encoding a neurotrophic factor (that is currently under clinical investigation and will be studied here) while additional preclinical studies could be followed to test the effectiveness of novel genetic targets in these advanced PD models. In addition, the approach could also be applied to other neurological disorders characterized by highly disseminated disease areas within the brain.