Characterization of the role of cyclin G-associated kinase in Parkinson disease

细胞周期蛋白 G 相关激酶在帕金森病中作用的表征

• 批准号: 8462710
• 负责人: RICHARD H MYERS
• 金额: $ 44.2万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8462710
• 关键词: Affect; Age; Age of Onset; Alcohol consumption; Alleles; Alternative Splicing; Alzheimer' s Disease; Bar Codes; Brain; Caffeine; Cathepsins; Cell model; Characteristics; Clinical Trials; Code; DNA Resequencing; DNA Sequence; Data; Dementia; Development; Diagnosis; Diagnostic; Dideoxy Chain Termination DNA Sequencing; Disease; Disease susceptibility; Endocytosis; Enzymes; Evaluation; Exhibits; Exons; Family history of; Family member; Frequencies; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Polymorphism; Genetic Risk; Genetic Transcription; Genomics; Genotype; Hand; Health; Hereditary Disease; Individual; Length; Mental Depression; Methods; Microarray Analysis; Molecular Profiling; Mutation; Neurodegenerative Disorders; Neurologic; Neurons; Parkinson Disease; Pathogenesis; Pathway interactions; Pattern; Phase; Phosphotransferases; Play; Population; Publishing; RNA; RNA Sequences; RNA Splicing; Reading; Recording of previous events; Relative (related person); Research; Research Personnel; Resolution; Risk; Risk Factors; Role; SNP genotyping; Sampling; Series; Signal Transduction; Smoking; Spliced Genes; Targeted Resequencing; Technology; Testing; Therapeutic; Therapeutic Human Experimentation; Therapeutic Intervention; Tissues; Toxic effect; Transcript; Tremor; Variant; alpha synuclein; base; case control; clinical risk; cohort; cyclin G; disorder control; disorder risk; exome sequencing; experience; frontal lobe; gene environment interaction; genome wide association study; genome-wide; insight; interest; mind control; next generation; novel; pesticide exposure; response; segregation; synuclein; transcriptome sequencing

DESCRIPTION (provided by applicant): Our genome wide association study (GWAS) for familial Parkinson's disease (PD) was the first to identify the cyclin G-associated kinase (GAK) gene as associated with PD risk. Meta-PD GWAS analyses have now demonstrated an unequivocal genome-wide effect (at least P = 4.8 x 10-15) for the rs1564282 SNP in the GAK gene on increased PD risk. Notably, the effect appears to be particularly strong in familial PD. Recently, we published that GAK is associated with 1-synuclein toxicity. Microarray expression analysis of post-mortem frontal cortex from PD and control brains found a significant association of rs1564282 with increased 1- synuclein expression, which has implications for disease pathogenesis. Further, knockdown of GAK significantly increases 1-synuclein toxicity in neuronal cell models of PD. GAK plays a critical role in endocytosis through its interaction with cathepsin-D (CTSD), which is the main lysosomal enzyme involved in 1-synuclein degradation. Taken together, these studies implicate a novel role for GAK in PD pathogenesis. Since kinases, such as GAK, are attractive targets for therapeutic intervention, resolving the responsible functional variants in the GAK gene region and their effects on GAK's expression and function promise important advancements for PD research and therapeutics. This application proposes to identify the responsible mutations, and to further evaluate their role in the implicated endocytic pathway and in the pathogenesis of PD. We propose a definitive series of DNA sequencing, RNA sequencing, alternative splicing, gene expression, and clinical risk profiling studies which have important translational implications. Specifically, Aim 1 will identify functional sequence variants in the GAK region by resequencing 225 Kb in 480 familial PD cases and 96 controls from our GenePD familial PD cohort, in which the gene was identified, thus greatly enhancing the likelihood for successful discovery of functional variants. Aim 2 proposes to perform RNA-sequencing in a large well characterized series of 34 PD and 29 control brains, for which extensive SNP genotyping and microarray data are already available. The RNA-Seq data will be used to study coding sequence and expression of GAK and related genes in the endocytic pathway. RNA sequencing will also allow us to examine all the GWAS implicated PD genes (e.g. SNCA, MAPT, BST1 etc.) and to perform transcriptome-wide comparison of cases and controls. Finally, Aim 3 explores the characteristics of PD cases carrying the variants and mutations found in Aims 1 and 2 and utilizes the genotyping of identified sequence variants in gene-gene and gene- environment interaction and genetic risk profiling studies. Genetic risk profiling studies have important translational implications in risk prediction and diagnostics for PD as well as for interpretation of therapeutic response in clinical trials for PD. These PD investigators are uniquely experienced with the GAK gene and region and have already in hand all the samples needed to carry out these definitive studies.

描述（由申请人提供）：我们的家族性帕金森病（PD）基因组全关联研究（GWAS）首次确定了细胞周期蛋白g相关激酶（GAK）基因与PD风险相关。Meta-PD GWAS分析现已证明GAK基因rs1564282 SNP对PD风险增加具有明确的全基因组效应（至少P = 4.8 x 10-15）。值得注意的是，这种影响在家族性帕金森病中表现得尤为明显。最近，我们发表了GAK与1-突触核蛋白毒性有关。对PD和对照脑的死后额叶皮层的微阵列表达分析发现，rs1564282与1-突触核蛋白表达增加有显著关联，这可能与疾病发病机制有关。此外，GAK的敲低显著增加PD神经元细胞模型中1-突触核蛋白的毒性。GAK通过与组织蛋白酶- d （CTSD）的相互作用在内吞作用中起关键作用，组织蛋白酶- d是参与1-突触核蛋白降解的主要溶酶体酶。综上所述，这些研究暗示了GAK在PD发病机制中的新作用。由于激酶，如GAK，是治疗干预的有吸引力的靶点，解决GAK基因区域的相关功能变异及其对GAK表达和功能的影响，为PD研究和治疗带来了重要的进展。本申请旨在鉴定相关突变，并进一步评估其在相关内吞途径和PD发病机制中的作用。我们提出了一系列明确的DNA测序、RNA测序、选择性剪接、基因表达和临床风险分析研究，这些研究具有重要的翻译意义。具体来说，Aim 1将通过重测序480例家族性PD病例和96例来自GenePD家族性PD队列的225 Kb基因来鉴定GAK区域的功能序列变异，从而大大提高成功发现功能变异的可能性。Aim 2提出在34个PD和29个对照脑中进行rna测序，其中广泛的SNP基因分型和微阵列数据已经可用。RNA-Seq数据将用于研究内吞途径中GAK及相关基因的编码序列和表达。RNA测序还将使我们能够检查所有与GWAS相关的PD基因（例如SNCA， MAPT， BST1等），并对病例和对照进行转录组范围的比较。最后，Aim 3探讨了携带Aims 1和2中发现的变异和突变的PD病例的特征，并在基因-基因和基因-环境相互作用以及遗传风险分析研究中利用鉴定的序列变异的基因分型。遗传风险分析研究对帕金森病的风险预测和诊断以及临床试验中治疗反应的解释具有重要的转化意义。这些PD研究人员对GAK基因和区域具有独特的经验，并且已经掌握了进行这些明确研究所需的所有样本。