Gene Delivery to the CNS through the Deep Cerebellar Nucleus

描述(由申请人提供)：为了以安全和高效的方式改善对中枢神经系统所有区域的基因输送，本提案的目标是开发、优化和测试这种替代输送途径，以克服血脑屏障的限制，这是本申请申请的一个具体重点。我们的策略集中在大脑的一个特殊区域，称为小脑深核(DCN)。我们将使用腺相关病毒载体(AAV)，这种载体已经被证明是一种非致病性的载体，在许多人类临床试验中已经建立了安全性描述，以及在帕金森氏病等疾病的临床成功的初步迹象，以及一种我们在1，2方面有相当经验的病毒载体。我们最近在肌萎缩侧索硬化症(ALS)的小鼠模型中研究了这种新的全球基因传递方法，该方法似乎是稳健的，因此是可行的。一些报告表明，神经营养因子有可能延缓疾病的发生和发展4-6。然而，将这些方法有效地转化为临床仍然存在挑战。我们推测，这种一次性向DCN注射的策略将导致全球CNS转导，从而允许对脑干、脊髓和运动皮质的广泛营养支持。我们进一步假设，当这种交付方法在ALS转基因小鼠身上进行测试时，这种方法将极大地提高运动协调性，延长生存期，并挽救神经病理症状。这项提议的主要目标是利用AAV载体为全球基因传递定义一条更有效的治疗路线。这种方法可能适用于治疗多种神经疾病和状况，主要与脊髓相关，如肌萎缩侧索硬化症和脊髓损伤，或与大脑有关，如阿尔茨海默氏症和帕金森病。我们的具体目标如下所示。1)目的：确定一次向DCN注射AAV是否针对中枢神经系统和脊髓的所有区域。2.)目的：研究DCN基因转导AAV-IGF-1对ALS动物模型的疗效。目前的基因治疗仍然是一个挑战，因为有有效的方法来靶向中枢神经系统，包括难以进入脊髓。系统地将基因治疗载体输送到中枢神经系统一直是具有挑战性的，主要是因为它们无法跨越血脑屏障(BBB)。我们的策略集中于利用腺相关载体(AAV)来靶向大脑的一个特殊区域，称为小脑深部核(DCN)，以传递转基因。我们决定以DCN为目标，因为它与脊髓和脑干的所有区域都有广泛的联系，因此将成为理想的基因传递途径。

DESCRIPTION (provided by applicant): In pursuit of improving gene delivery to all areas of the CNS in a safe and efficient manner, the objective of this proposal is to develop, optimize, and test this alternate delivery route to overcome the limitations of the blood brain barrier, which is a specific focus of this Request for Applications. Our strategy is centered on a specialized region of the brain termed the deep cerebellar nuclei (DCN). We will employ Adeno- Associated Viral Vector (AAV), a vector that has shown to be non-pathogenic with safety profiles being established in numerous human clinical trials as well as initial signs of clinical success in diseases such as Parkinson's disease and a viral vector that we have considerable experience with 1, 2 . We have recently examined this novel global gene delivery approach in a mouse model for Amyotrophic Lateral Sclerosis (ALS) with the approach appearing to be robust and therefore feasible. Several reports have shown the potential actions of neurotrophic factors to delay disease onset and progression 4-6 . However, challenges still remain to effectively translate these approaches to the clinic. We hypothesize that this one-time single injection delivery strategy to the DCN will result in global CNS transduction, allowing for widespread trophic support to the brainstem, spinal cord and motor cortex. We further hypothesize that this approach will greatly increase motor coordination, extend survival, and rescue neuropathological symptoms when the delivery approach is tested in ALS transgenic mice. The main goal of this proposal is to define a more effective therapeutic route for global gene delivery utilizing an AAV vector. This approach may be amenable for treating multiple neurological diseases and conditions, primarily associated either with the spinal cord such as ALS and spinal cord injury, or with the brain such as Alzheimer's and Parkinson's Disease. Our specific aims are shown below. 1.) To determine whether a single injection of AAV to the DCN targets the CNS and all regions of the spinal cord. 2.) To determine the efficacy of DCN gene delivery of AAV-IGF-1 in an animal model of ALS. Current gene therapy remains to be a challenge due to effective methods to target the CNS including the difficulty to access the spinal cord. Systemic delivery of gene therapy vectors to the CNS has been challenging due largely by their inability to cross the blood-brain barrier (BBB). Our strategy is centered on targeting a specialized region of the brain termed the deep cerebellar nuclei (DCN) with adeno-associated vectors (AAV) to deliver transgenes. We have decided to target the DCN because of its extensive connections with all regions of the spinal cord and brainstem, and would thus serve as an ideal gene delivery route.