Role of Inflammation and Oxidative Stress in Parkinson's Disease

描述（由申请人提供）： 这项拨款提案的重点的总体假设是，慢性TH 1/经典炎症对帕金森病（PD）和海湾战争疾病（GWI）和其他神经退行性疾病的神经退行性疾病的进展至关重要。文献中有强有力的证据表明，在PD患者、GWI患者和患有其他神经退行性疾病的患者的大脑中观察到炎症和氧化损伤，然而关键问题是神经元、神经胶质和T细胞的相互作用。此外，有越来越多的证据表明，在衰老期间，存在不利环境的发展，这使得可能对年轻动物有效的治疗剂在老年动物中不太有效。本提案的目的是通过跟踪黑质中过表达的野生型（WT）α-突触核蛋白（通过腺相关病毒（AAV）9转导）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，并确定在年轻动物中有效的治疗方法是否在老年动物中同样有效。 具体目标1：假设：CX 3CL 1（fractalkine）是一种在神经胶质细胞相互作用中重要的趋化因子。我们提出给予CX 3CL 1的基因治疗方法是可行的，并且将有效对抗PD的rAAV 9-突触核蛋白模型。我们将进一步研究趋化因子信号转导和T细胞通过IL 12的相互作用。CX 3CL 1是神经胶质细胞通讯中重要的细胞因子，然而关于哪种形式的CX 3CL 1具有神经保护作用存在争议，因为有神经退行性作用的报道。我们将使用具有差异加工的CX 3CL 1的3种AAV 9构建体继续探索这个问题。此外，α-SYN模型中正在进行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。我们有初步的数据表明，CX 3CL 1的一个作用是减少IL 12，这是TH 1免疫的重要调节因子。 具体目标2：假设：衰老会改变先天性和适应性免疫系统，因此会改变基于免疫的治疗的疗效。由于大多数PD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AAV 9-CX 3CL 1疗法在老年动物的AAV 9-α-SYN模型PD中是否有效。我们知道，老年啮齿动物的小胶质细胞主要表现为M1反应，并减弱了M2信号。这可以改变神经元-胶质细胞-T细胞趋化因子信号传导。我们将研究CX 3CL 1治疗改变小胶质细胞反应的效果，并与T细胞群的调节相互作用。 特定目的3：当T细胞存在时，α-突触核蛋白神经毒性增加，并且依赖于T细胞-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细胞将改变迁移、归巢和存活的模式。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重要作用。这尚未在涉及α-突触核蛋白的PD模型中进行广泛研究，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.