Role of pathogenic Parkinsonian mutations in the seeding and propagation of alpha-synuclein in the CNS

摘要 帕金森氏病的病因是多元的，从已发现的基因突变到 严格的环境因果关系。到目前为止，已经确定了18个以上导致帕金森症的基因。 导致帕金森氏症的两个最常见的基因突变是：1)GBA基因突变， 编码葡萄糖脑苷酶蛋白，以及2)富含亮氨酸重复的LRRK2基因突变 蛋白激酶II。除了已知的“遗传即家族”与疾病病因的关系外，两者 GBA和LRRK2基因也被认为是帕金森病发生的危险因素，因为 每个携带这些突变的人都会患上帕金森氏症，可能只会在一秒钟后显现出来 “Hit”。无论帕金森病的起因是什么，几乎所有的帕金森氏病病例都有共同的方面 病理改变，包括：1)聚集的α-突触核蛋白的存在，2)SNPC DA神经元的丢失，3) 神经炎症的增加。此外，人们还可以看到认知和运动输出的变化。在这 应用程序，我们将检查已知的引发帕金森氏病的不同病理机制， 包括蛋白激酶激活、蛋白管理或炎症将改变/影响聚集 以及α-SYN在整个神经系统中的传播。具体地说，我们将检查对PD的影响 病理生理学包括SNPC DA神经元丢失，基底节儿茶酚胺丢失，诱导 神经炎症和错误折叠的α-突触核蛋白扩散。我们还将检查认知和运动能力是否 在PFF播种后，这3种情况下都会发生行为变化。这些帕金森氏症的病理将是 将预制的α-突触核蛋白微丝注射到三个不同区域后的检查 包括两个已知参与帕金森病(嗅球和纹状体)和一个没有参与的中枢神经系统 (内部控制，海马体)。在具体目标1中，我们将检查是否将PFF注入不同的区域 携带LRRK2基因G2019S突变的小鼠中枢神经系统改变α-SYN的播种和传播 以及改变如上所述的PD中的其他已知病理。在具体目标2中，我们将检查是否 α-突触核蛋白的预成纤维注射到携带AFP的小鼠中枢不同区域 L444P GBA突变改变α-SYN的播种和传播，以及改变其他已知的病理机制 如上所述的PD。在特定的目标3中，我们将测试假设先前的神经炎性侮辱 (感染H1N1流感病毒)到大脑会增加pff在小鼠体内的播种和传播 携带帕金森病易感基因，以及改变如上所述的帕金森病的其他已知病理。这些 三个目标将使我们能够确定这些病理机制中的任何一个或多个(激酶激活 (遗传)、蛋白质处理不当(基因x环境)或病毒感染(环境)直接影响 错误折叠的α-突触核蛋白和其他常见帕金森病的传播。

Abstract The etiology of Parkinson’s disease is multivariate, ranging from identified genetic mutations to strict environmental causation. So far, more than 18 genes have been identified that result in parkinsonism. The two most common genetic mutations that lead to parkinsonism are: 1) mutations in the GBA gene that encodes the glucocerebrosidase protein, and 2) mutations in the LRRK2 gene that Leucine Rich Repeat Kinase II protein. In addition to their known “genetic i.e familial” relationship to disease causation, both the GBA and LRRK2 genes are also considered to be “risk factors” for development of PD, in that not everyone with these mutations develops Parkinson’s disease and they may only manifest after a second “hit”. No matter the initiating cause of PD, almost all cases of Parkinson’s disease share common aspects of pathology, including: 1) the presence of aggregated alpha-synuclein, 2) loss of SNpc DA neurons and 3) an increase in neuroinflammation. Additionally, one also sees cognitive and motor output changes. In this application, the we will examine different pathological mechanisms known to initiate Parkinson’s disease, including protein kinase activation, protein management or inflammation will alter/affect the aggregation and spread of α-syn throughout the nervous system. Specifically, we will examine the effect on PD pathophysiology including SNpc DA neuron loss, loss of basal ganglia catecholamines, induction of neuroinflammation and spread of misfolded alpha-synuclein. We will also examine if cognitive and motor behavioral changes occur in these 3 conditions after PFF seeding. These parkinsonian pathologies will be examined following injection of preformed filaments of alpha-synuclein (PFFs) into three different regions of the CNS, including two known to be involved in PD (olfactory bulb and striatum) and one that is not (internal control, hippocampus). In Specific Aim 1, we will examine if PFFs injected into different regions of the CNS of mice carrying a G2019S mutation in the LRRK2 gene alter the seeding and spread of α-Syn as well as alter other known pathologies in PD as described above. In Specific Aim 2, we will examine if preformed fibrils of alpha-synuclein (PFFs) injected into different regions of the CNS of mice carrying a L444P GBA mutation alters the seeding and spread of α-Syn as well as alter other known pathologies in PD as described above. In Specific Aim 3 we will test the hypothesis that a prior neuroinflammatory insult (infection with the H1N1 influenza virus) to the brain will increase the seeding and spread of PFFs in mice carrying PD susceptibility genes as well as alter other known pathologies in PD as described above. These three aims will allow us to determine if any one or more of these pathological mechanisms (kinase activation (genetic), protein mishandling (gene x environment” or viral infection (environment) directly influence the spread of misfolded alpha-synuclein and other common parkinsonian pathologies.