Role of Inflammation and Oxidative Stress in Parkinson's Disease

描述（由申请人提供）： 本拨款提案重点的总体假设是，慢性 TH1/经典炎症对于帕金森病 (PD) 和海湾战争疾病 (GWI) 以及其他神经退行性疾病的神经退行性疾病进展至关重要。文献中有强有力的证据表明，在 PD 患者、GWI 患者和其他神经退行性疾病患者的大脑中观察到炎症和氧化损伤，但一个关键问题是神经元、胶质细胞和 T 细胞的相互作用。此外，越来越多的证据表明，在衰老过程中，会形成一种不利的环境，使得对年轻动物可能有效的治疗方法对老年动物效果较差。该提案的目的旨在通过腺相关病毒 (AAV)9 转导跟踪黑质中过度表达的野生型 (WT) α-突触核蛋白的损伤进展来检查这一过程。我们将研究炎症在该模型中的作用，并确定对幼年动物有效的治疗方法是否对老年动物同样有效。 具体目标 1：假设：CX3CL1（fractalkine）是一种在神经元胶质相互作用中重要的趋化因子。我们提出，施用 CX3CL1 的基因治疗方法是可行的，并且对 PD 的 rAAV9-突触核蛋白模型有效。我们将进一步研究趋化因子信号传导与 T 细胞通过 IL12 的相互作用。 CX3CL1 是神经胶质细胞通讯中重要的细胞因子，然而，由于有神经退行性作用的报道，关于哪种形式的 CX3CL1 具有神经保护作用存在争议。我们将使用具有差异处理的 CX3CL1 的 3 个 AAV9 构建体继续探索这个问题。此外，α-SYN 模型中持续的神经变性涉及调节先天免疫细胞和 T 细胞的趋化因子信号的复杂相互作用。我们有初步数据表明CX3CL1的作用之一是减少IL12，IL12是TH1免疫的重要调节剂。 具体目标 2：假设：衰老会改变先天性和适应性免疫系统，从而改变免疫疗法的功效。由于帕金森病大多数发生在老年受试者中，因此在治疗选择方面考虑这一点至关重要。我们将检查 AAV9-CX3CL1 疗法在老年动物的 AAV9-α-SYN 模型 PD 中是否有效。我们知道，老年啮齿类动物的小胶质细胞主要表现出 M1 反应，并减弱了 M2 信号传导。这可能会改变神经元 - 胶质细胞 - T 细胞趋化因子信号传导。我们将检查 CX3CL1 治疗对改变小胶质细胞反应以及与 T 细胞群调节相互作用的影响。 具体目标 3：当 T 细胞存在时，α-突触核蛋白的神经毒性会增加，并且依赖于 T 细胞与小胶质细胞通过趋化因子的通讯。我们进一步预测老年大鼠的 T 细胞的迁移、归巢和生存模式将发生改变。越来越多的证据表明，T 细胞与小胶质细胞和其他抗原呈递细胞一起在神经变性中发挥着重要作用。这在涉及 α-突触核蛋白的 PD 模型中尚未得到广泛研究，但一项研究表明存在相关性。我们将在无胸腺裸鼠中对此进行研究，然后通过过继转移方法单独替换 T 细胞亚群来确定涉及哪种类型的 T 细胞。

DESCRIPTION (provided by applicant): The overall hypothesis that underlies the focus of this grant proposal is that chronic TH1/classical inflammation is critical for the progression of neurodegeneration in Parkinson's disease (PD) and Gulf War Illnesses (GWI) and other neurodegenerative diseases. There is strong evidence from the literature that inflammation and oxidative damage are observed in the brains of PD patients, GWI patients and patients with other neurodegenerative diseases, however a critical question is the interaction of neurons, glia and T cells. Furthermore, there is mounting evidence that an during aging, there is development of a hostile environment that makes therapeutics that may be effective in the young animal, less effective in the aged animal. The aims of this proposal are designed to examine this process by following the progression of damage from wild-type (WT) α-synuclein overexpressed in the substantia nigra via transduction with adeno-associated virus (AAV)9. We will study the role of inflammation in this model and determine if therapeutics effective in young animals are equally effective in the aged. Specific Aim 1: Hypothesis: CX3CL1 (fractalkine) is a chemokine important in neuronal glial interactions. We propose that a gene therapy approach to administering CX3CL1 is viable and will be effective against rAAV9-synuclein models of PD. We will further examine interactions of chemokine signaling and T cells via IL12. CX3CL1 is a cytokine important in neuronal glial communication, however there is debate concerning which form of CX3CL1 is neuroprotective, as there are reports of neurodegenerative actions. We will continue exploring this question using 3 AAV9 constructs of CX3CL1 that have differential processing. In addition, ongoing neurodegeneration in α-SYN models involves a complex interaction of chemokine signals that regulate both innate immune cells and T cells. We have preliminary data that one action of CX3CL1 is to reduce IL12, and important regulator of TH1 immunity. Specific Aim 2: Hypothesis: Aging alters both the innate and adaptive immune system, thus will alter the efficacy of immune based therapies. As PD the majority of PD occurs in elderly subjects, it is critical to consider this with respect to therapeutic choices. We will examine if AAV9-CX3CL1 therapy is effective in the AAV9-α-SYN model PD in aged animals. WE know that microglia in aged rodents show predominantly M1 responses and have blunted M2 signaling. This may alter neuronal - glial-Tcell chemokine signaling. We will examine the effect of CX3CL1 treatment to alter microglial responses and interact with regulation of T cell populations. Specific Aim 3: α-Synuclein neurotoxicity is increased when T cells are present and depends on T cell- microglia communication via chemokines. We further predict that T cells in aged rats will have altered patterns of migration, homing and survival. Evidence is mounting that T cells play an important role in neurodegeneration in concert with microglia and other antigen presenting cells. This has not been studied extensively in models of PD involving α-synuclein, however one study suggests a correlation. We will investigate this in the athymic nude rat, and then determine which type of T cell is involved by replacing T cell subsets individually by adoptive transfer methods.