Investigating the role of MARKs in Parkinson's disease pathogenesis

神经退行性疾病是老年人口衰弱的主要原因，但没有改变疾病的方法。 有治疗方法可以延缓疾病的发展。α-突触核蛋白在病理性聚集体中积聚 在帕金森氏症、路易体痴呆和约50%的阿尔茨海默病患者的大脑中。 α-突触核蛋白在这种疾病的家族性形式中也会发生突变。然而，人们对其致病途径知之甚少。 潜在的联核病，导致治疗靶点的匮乏。更好的理解提供了 通过对这一途径的分子组成的系统分析，将提供可能被利用的靶标 使用假设驱动的临床干预。这里提出的实验将利用一种 已建立的细胞模型，除了新技术外，还概括了联核病的主要特征 解剖最近发现的微管亲和力的分子途径的分子成分 调节蛋白激酶(MARKs)是α-突触核蛋白发病机制中的重要调节因子。MARKs在激活残基处被磷酸化，并在突触核病的细胞和动物模型以及人脑的路易体中聚集。抑制MARs可加重α-突触核蛋白的病理改变，提示MARS可能对α-突触核蛋白诱导的发病有保护作用。这项提议的长期目标是探索这样一种假设，即标记及其效应物和底物是α-突触核蛋白致病途径的组成部分，可以通过对相关蛋白质的治疗靶向来开发。这一假设将通过两个具体目标进行检验。在目标1中，将利用CRISPRi/a平台选择性地增加或减少标记效应物和底物的表达，以确定这些蛋白如何影响α-突触核蛋白的病理。结合对MARK活性的操作，该检测将用于确定α-突触核蛋白致病级联反应的蛋白质组分。在目标2中，我们将结合免疫细胞化学和活体成像研究MARK活性与错误折叠的α-突触核蛋白的自噬靶向之间的联系，以确定MARK对α-突触核蛋白的轴突运输和毒性的影响。 病理学。这项拟议的工作将为α-突触核蛋白病理如何形成和识别提供有价值的见解 控制这一过程的蛋白质，目的是开发这些蛋白质用于治疗帕金森病和 相关的联体核病。

Neurodegenerative diseases are a primary cause of debilitation in the aging population, yet no disease-modifying treatments are available to slow the progression of disease. α-Synuclein accumulates in pathological aggregates in the brains of patients with Parkinson’s disease, dementia with Lewy bodies and ~50% of Alzheimer’s disease. α-Synuclein is also mutated in familial forms of the disease. Yet, little is known about the pathogenesis pathway underlying synucleinopathies, leading to a paucity of therapeutic targets. The improved understanding provided by systematic analysis of the molecular components of this pathway will provide targets that may be exploited using hypothesis-driven clinical interventions. The experiments proposed here will take advantage of an established cell model that recapitulates major features of synucleinopathies, in addition to novel technologies to dissect the molecular components of a recently identified molecular pathway in which microtubule-affinity regulating kinases (MARKs) are important modulators of α-synuclein pathogenesis. MARKs are phosphorylated at an activating residue and accreted in Lewy bodies of cell and animal models of synucleinopathy as well as human brains. Inhibition of MARKs leads to an exacerbation of α-synuclein pathology, suggesting that MARKs may protect against α-synuclein-induced pathogenesis. The long-term objective of this proposal is to explore the hypothesis that MARKs, their effectors and substrates are components of an α-synuclein pathogenesis pathway that can be exploited through therapeutic targeting of the proteins involved. The hypothesis will be tested through two specific aims. In Aim 1, a CRISPRi/a platform will be utilized to selectively increase or decrease expression of MARK effectors and substrates to determine how these proteins affect α-synuclein pathology. Together with manipulation of MARK kinase activity this assay will be used to define protein components of the α-synuclein pathogenesis cascade. In Aim 2, the link between MARK activity and the targeting of misfolded α-synuclein for autophagy will be explored using a live imaging paradigm in parallel with immunocytochemistry to determine the effect MARKs have on the axonal transport and toxicity of α-synuclein pathology. The proposed work will offer valuable insight into how α-synuclein pathology is formed and identify proteins that control this process with the goal of developing those proteins for therapeutic treatment of PD and related synucleinopathies.