Investigating Neuroprotective Targets for Parkinson Disease Associated with Autop

研究与 Autop 相关的帕金森病的神经保护目标

• 批准号: 8180154
• 负责人: Guy A Caldwell
• 金额: $ 41.15万
• 依托单位: UNIVERSITY OF ALABAMA IN TUSCALOOSA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8180154
• 关键词: Acute; Address; Age; Aging; American; Animal Model; Apoptosis; Attenuated; Autophagocytosis; Axon; Biological Assay; Biological Models; Caenorhabditis elegans; Candidate Disease Gene; Clinical; Data; Deletion Mutation; Dependence; Dependency; Development; Diagnosis; Disease; Dopamine; Dose; Environment; Evaluation; Gene Proteins; Gene Targeting; Genes; Genetic; Genetic Polymorphism; Genomics; Goals; Human; Human Genome; Incidence; Individual; Insulin; Insulin Signaling Pathway; Link; Longevity; Mediating; Medical; Modeling; Molecular; Movement Disorders; Mutation; Nematoda; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Onset of illness; Orthologous Gene; Parkinson Disease; Pathway interactions; Patients; Phosphotransferases; Play; Protein-Serine-Threonine Kinases; Proteins; RNA Interference; Reporting; Research; Risk Factors; Role; Science; Scientist; Screening procedure; Series; Signal Pathway; Single Nucleotide Polymorphism; Structure; Students; System; Therapeutic; Training; Transgenic Organisms; age related; alpha synuclein; combat; design; dopaminergic neuron; functional genomics; genome wide association study; graduate student; in vivo; inhibition of autophagy; inhibitor/antagonist; insulin signaling; knock-down; mutant; neuron loss; neuronal survival; neuroprotection; new therapeutic target; programs; protein misfolding; research study; synuclein; therapeutic development

DESCRIPTION (provided by applicant): Therapeutic advances to treat Parkinson disease (PD) are incumbent upon the identification and mechanistic understanding of cellular factors that influence neuronal survival. We have exploited attributes of the nematode model system, Caenorhabditis elegans, to develop assays that facilitate the screening and isolation of conserved genetic factors that mediate clinical hallmarks of PD, including intracellular aggregation of a-synuclein (a-syn) and dopamine (DA) neuron loss. This R15 application extends prior studies where we have identified neuroprotective targets for translational development and established genetic platforms to evaluate functional modifiers of age-dependent neurodegeneration. Increasing evidence is mounting for a significant role for lysosomal function as a mechanism influencing PD. In this context, a focal point of this application is the ULK2 protein - a human ortholog of the worm unc-51 gene product, a kinase implicated in autophagy, as well as axon elongation and guidance. We previously uncovered a neuroprotective activity for ULK2 in enhancing DA neuron survival in transgenic nematodes. Our data took on greater significance following the report of a human genome-wide association study that also identified a polymorphism in ULK2 as being associated with PD patients. We propose to advance our understanding of ULK2-mediated neuroprotection through a series of structure-function analyses, as well as evaluation of modifiers of ULK2, to mechanistically define the role of ULK2 kinase activity in attenuating neurodegeneration. C. elegans is also among the best-understood animal models in terms of aging mechanisms, with extensive sets of factors implicated in lifespan identified. Thus, as aging represents an unequivocal and defined risk factor for PD, we will use functional genomic screening via RNA interference (RNAi) in C. elegans to knockdown hundreds of genes previously linked to aging-associated pathways to evaluate their distinct contribution to a-syn misfolding and clearance. Preliminary studies in our lab have shown that specific mutations in key components of the daf-2/insulin-like signaling pathway of C. elegans result in substantial effects on DA neuron survival. We have generated a series of transgenic nematode strains to facilitate functional analysis of a-syn modifiers in the context of their potential dependence on this pathway, as well as in autophagy. Transgenic and mutant worms will be generated to examine gene targets from RNAi screening for their impact on DA neuroprotection. This systematic approach provides an unprecedented opportunity to discern age-associated regulators of neurodegeneration that may represent genetic susceptibly markers for PD onset or progression. Collectively, these studies represent an integrated research plan designed to rapidly define the significance of previously uncharacterized factors influencing neurodegeneration. Moreover, our experimental strategy coincides with the specific criteria of the R15 AREA program, as the broader impacts of this application involve extensive undergraduate and graduate student training opportunities in an environment ideally suited to student-centered research. PUBLIC HEALTH RELEVANCE: Over 1 million Americans have been diagnosed with Parkinson's Disease (PD) - the most common movement disorder for which a cure has eluded medical science for decades. This application addresses an unmet challenge of discerning genetic factors that may contribute to this neurodegenerative disease, while accelerating our understanding of underlying cellular mechanisms of PD. In this research, we use a simple animal model system to explore the well-established but poorly understood relationship of aging to PD, in addition to genetic factors that mediate the survival of dopamine-producing neurons. The experimental strategy outlined integrates student-centered research in the context of a systematic approach involving genetic and genomic analysis to uncover mechanisms that facilitate identification of new therapeutic targets with the potential to combat PD.

描述（由申请人提供）：帕金森病（PD）的治疗进展是对影响神经元存活的细胞因子的识别和机制理解的责任。我们利用线虫模型系统秀丽隐杆线虫（Caenorhabditis elegans）的属性，开发了有助于筛选和分离PD临床特征的保守遗传因子的检测方法，包括细胞内a-突触核蛋白（a-syn）聚集和多巴胺（DA）神经元丢失。这项R15应用扩展了之前的研究，我们已经确定了翻译发展的神经保护靶点，并建立了遗传平台来评估年龄依赖性神经变性的功能调节剂。越来越多的证据表明，溶酶体功能在影响PD的机制中起着重要作用。在这种情况下，该应用的焦点是ULK2蛋白-蠕虫unc-51基因产物的人类同源物，一种涉及自噬以及轴突延伸和引导的激酶。我们之前发现了ULK2在提高转基因线虫DA神经元存活方面的神经保护活性。在一项人类全基因组关联研究报告之后，我们的数据具有更大的意义，该研究也确定了ULK2多态性与PD患者相关。我们建议通过一系列的结构-功能分析，以及对ULK2调节剂的评估，来推进我们对ULK2介导的神经保护的理解，从而从机制上确定ULK2激酶活性在减轻神经退行性变中的作用。就衰老机制而言，秀丽隐杆线虫也是最容易理解的动物模型之一，已经确定了与寿命有关的广泛因素。因此，由于衰老是帕金森病的一个明确的危险因素，我们将通过RNA干扰（RNAi）在秀丽隐杆线虫中使用功能基因组筛选来敲除数百个先前与衰老相关途径相关的基因，以评估它们对a-syn错误折叠和清除的独特贡献。我们实验室的初步研究表明，秀丽隐杆线虫daf-2/胰岛素样信号通路关键组分的特异性突变对DA神经元的存活有实质性影响。我们已经产生了一系列的转基因线虫菌株，以促进a-syn修饰剂在其潜在依赖于该途径以及自噬的背景下的功能分析。将产生转基因和突变的蠕虫，以检测RNAi筛选的基因靶点对DA神经保护的影响。这种系统的方法提供了一个前所未有的机会来辨别神经退行性变的年龄相关调节因子，这些调节因子可能代表PD发病或进展的遗传易感标记。总的来说，这些研究代表了一个综合的研究计划，旨在快速定义以前未表征的影响神经变性的因素的重要性。此外，我们的实验策略与R15区域计划的具体标准相吻合，因为该应用程序的更广泛影响涉及广泛的本科生和研究生培训机会，非常适合以学生为中心的研究环境。

项目成果

NCEH-1 modulates cholesterol metabolism and protects against α-synuclein toxicity in a C. elegans model of Parkinson's disease.

NCEH-1 在帕金森病的秀丽隐杆线虫模型中调节胆固醇代谢并防止 α-突触核蛋白毒性。

• DOI: 10.1093/hmg/ddx269
• 发表时间: 2017
• 期刊: Human molecular genetics
• 影响因子: 3.5
• 作者: Zhang,Siyuan;Glukhova,SamanthaA;Caldwell,KimA;Caldwell,GuyA
• 通讯作者: Caldwell,GuyA

Valproic acid ameliorates C. elegans dopaminergic neurodegeneration with implications for ERK-MAPK signaling.

• DOI: 10.1016/j.neulet.2013.02.026
• 发表时间: 2013-04-29
• 期刊: Neuroscience letters
• 影响因子: 2.5
• 作者: Kautu BB;Carrasquilla A;Hicks ML;Caldwell KA;Caldwell GA
• 通讯作者: Caldwell GA

The glycolytic enzyme, GPI, is a functionally conserved modifier of dopaminergic neurodegeneration in Parkinson's models.

• DOI: 10.1016/j.cmet.2014.04.017
• 发表时间: 2014-07-01
• 期刊: Cell metabolism
• 影响因子: 29
• 作者: Knight AL;Yan X;Hamamichi S;Ajjuri RR;Mazzulli JR;Zhang MW;Daigle JG;Zhang S;Borom AR;Roberts LR;Lee SK;DeLeon SM;Viollet-Djelassi C;Krainc D;O'Donnell JM;Caldwell KA;Caldwell GA
• 通讯作者: Caldwell GA