Role of Inflammation and Oxidative Stress in Parkinson's Disease

描述(由申请人提供)： 这项赠款提案的重点所在的总体假设是，慢性TH1/经典炎症对帕金森氏病(PD)、海湾战争病(GWI)和其他神经退行性疾病的神经退行性变的进展至关重要。有强有力的证据表明，PD患者、GWI患者和其他神经退行性疾病患者的大脑中都存在炎症和氧化损伤，但一个关键的问题是神经元、胶质细胞和T细胞之间的相互作用。此外，越来越多的证据表明，在衰老过程中，会发展出一种敌对的环境，使得对幼年动物有效的治疗方法在老年动物身上效果不佳。这项建议的目的是通过跟踪野生型(WT)α-突触核蛋白通过转导腺相关病毒9在黑质过度表达所造成的损害的进展来研究这一过程。我们将研究炎症在这一模型中的作用，并确定在幼年动物中有效的治疗方法是否在老年动物中同样有效。特定目的1：假设：CX3CL1(Fractalkine)是一种在神经元胶质细胞相互作用中起重要作用的趋化因子。我们认为，使用CX3CL1的基因治疗方法是可行的，并将对rAAV9-突触核蛋白帕金森病模型有效。我们将进一步研究趋化因子信号和T细胞通过IL12的相互作用。CX3CL1是一种在神经胶质细胞通讯中起重要作用的细胞因子，但关于哪种形式的CX3CL1具有神经保护作用还存在争议，因为有报道称CX3CL1具有神经退行性作用。我们将使用具有差异处理的3个CX3CL1的AAV9构造继续探索这个问题。此外，α-SYN模型中正在进行的神经退变涉及一种复杂的趋化因子信号相互作用，这些信号既调节先天免疫细胞又调节T细胞。我们有初步的数据表明，CX3CL1的作用之一是降低IL-12，这是TH1免疫的重要调节因子。具体目标2：假设：衰老改变先天免疫系统和获得性免疫系统，从而改变基于免疫的疗法的疗效。由于帕金森病大多数发生在老年受试者中，因此在治疗选择上考虑这一点是至关重要的。我们将检验AAV9-CX3CL1治疗对老年动物AAV9-α-SYN模型帕金森病是否有效。我们知道，老年啮齿动物的小胶质细胞主要表现为M1反应，并削弱了M2信号。这可能会改变神经元-神经胶质-T细胞趋化因子信号。我们将研究CX3CL1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。具体目的3：当T细胞存在时，α-突触核蛋白的神经毒性增加，并依赖于T细胞-小胶质细胞通过趋化因子的通讯。我们进一步预测，衰老大鼠的T细胞的迁移、归巢和存活模式将发生改变。越来越多的证据表明，T细胞与小胶质细胞和其他抗原提呈细胞在神经退行性变中发挥着重要作用。在涉及α-突触核蛋白的帕金森病模型中，这一点还没有得到广泛的研究，然而一项研究表明了这种相关性。我们将在裸鼠身上研究这一点，然后通过采用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.