Gene Delivery to the CNS through the Deep Cerebellar Nucleus

描述（由申请人提供）：为了以安全有效的方式改善基因递送到中枢神经系统的所有区域，本提案的目标是开发、优化和测试这种替代递送途径，以克服血脑屏障的限制，这是本申请的一个特定重点。我们的策略集中在大脑的一个特殊区域，称为小脑深部核（DCN）。我们将使用腺相关病毒载体（AAV），这是一种在许多人体临床试验中证明具有非致病性和安全性的载体，并且在帕金森病等疾病中取得了初步临床成功的迹象，也是一种我们有丰富经验的病毒载体1,2。我们最近在肌萎缩性侧索硬化症（ALS）小鼠模型中研究了这种新的全局基因传递方法，该方法似乎是稳健的，因此是可行的。一些报告显示神经营养因子的潜在作用可以延缓疾病的发生和进展4-6。然而，将这些方法有效地转化为临床仍然存在挑战。我们假设，这种一次性的单次注射给药策略将导致中枢神经系统的全局性转导，从而允许对脑干、脊髓和运动皮层的广泛营养支持。我们进一步假设，当在ALS转基因小鼠中测试这种递送方法时，这种方法将大大提高运动协调能力，延长生存期，并挽救神经病理症状。本提案的主要目标是利用AAV载体定义一种更有效的全球基因传递治疗途径。这种方法可能适用于治疗多种神经系统疾病和病症，主要与脊髓有关，如ALS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。1.） 确定单次向DCN注射AAV是否靶向中枢神经系统和脊髓的所有区域。2)。目的：探讨AAV-IGF-1在ALS动物模型中的DCN基因转染效果。目前的基因治疗仍然是一个挑战，因为没有有效的方法来靶向中枢神经系统，包括难以进入脊髓。由于基因治疗载体无法通过血脑屏障（BBB），将其全身递送到中枢神经系统一直具有挑战性。我们的策略集中在用腺相关载体（AAV）靶向被称为小脑深部核（DCN）的大脑特殊区域来传递转基因。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，因此可以作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.