Exploring Disease-Toxicant Interactions in a Human Huntington's Disease Model

探索人类亨廷顿病模型中的疾病与毒性相互作用

• 批准号: 8670787
• 负责人: Andrew M Tidball
• 金额: $ 0.72万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8670787
• 关键词: Affect; Age; Analysis of Covariance; Basic Science; Biological Assay; Biological Models; Brain; Brain region; CAG repeat; Calcium; Cell Culture Techniques; Cell Death; Cell Line; Cell Survival; Cells; Cellular Stress; Cellular Stress Response; Codon Nucleotides; Communities; Complex; Corpus striatum structure; Data; Development; Disease; Disease Progression; Disease model; Environment; Environmental Health; Environmental Medicine; Environmental Risk Factor; Etiology; Exhibits; Exposure to; Freedom; Functional disorder; Funding; Generations; Genes; Genetic; Genetic Risk; Genotype; Goals; Heat shock proteins; Human; Huntington Disease; In Vitro; Individual; Inheritance Patterns; Juvenile-Onset Huntington Disease; Knowledge; Laboratories; Left; Length; Measures; Membrane Potentials; Messenger RNA; Metabolic; Metals; Mitochondria; Modeling; Modification; Multivariate Analysis; Mutate; Nerve Degeneration; Neurodegenerative Disorders; Neurologic; Neuronal Dysfunction; Neurons; Neurotoxins; Other Genetics; Ownership; Oxidative Stress; Pathway interactions; Patients; Phenotype; Population; Predisposition; Principal Investigator; Process; Production; Promoter Regions; Proteins; Protocols documentation; Reactive Oxygen Species; Regression Analysis; Research; Research Personnel; Research Project Grants; Risk; Staging; Statistical Models; Stress; Study models; Techniques; Testing; Toxic Environmental Substances; Toxic effect; Toxicant exposure; Training; Variant; base; cell type; design; disorder control; environmental agent; environmental stressor; excitotoxicity; genetic risk factor; human Huntingtin protein; human subject; induced pluripotent stem cell; inhibitor/antagonist; mitochondrial dysfunction; mitochondrial membrane; mutant; nerve stem cell; neuropathology; regional difference; research study; response; stress protein; stressor; therapy development; toxicant; toxicant interaction

DESCRIPTION (provided by applicant): Environmental agents are thought to interact with an individual's genetic makeup to influence or cause neurodegenerative disease. Huntington's disease (HD) is a debilitating neurological condition with a dominant monogenic inheritance pattern. Still, environmental factors are thought to strongly affect disease age at onset and progression. We postulate that the environment causes cellular stress that neurons in the HD brain are susceptible to such as oxidative stress, mitochondrial dysfunction, and calcium dysregulation. We hypothesize that mutant Huntingtin impinges upon the cellular stress response to produce this disease-specific vulnerability. We will test this hypothesis by exposing cells to toxicants known to elicit implicated cellular stress pathways and measure toxicological phenotypes. Here we propose using induced pluripotent stem cells differentiated into striatal neural progenitors from patients with Huntington's disease and controls as our model system. These cell lines contain the mutated Huntingtin gene with the complete promoter region as well as patient- specific genetic background in a neural cell type. In Specific Aim 1, we will optimize the technique for generating DLX2-expressing neural progenitors. We will then expose DLX2-expressing cultures from hiPSCs from two juvenile HD subjects and two control subjects to cell stress model toxicants and evaluate for differential viability. In Specific Aim 2, these same cells will be used to assay the effects toxicant exposure on reactive oxygen species production, mitochondrial membrane potential, ATP content, and cytosolic calcium concentration. The data generated in Aim 1 and Aim 2 will be analyzed by multivariate ANCOVA statistical modeling for disease-toxicant and/or patient-toxicant interactions. For toxicants eliciting a statistically significant disease-toxicant interaction, we will then perform protein and mRNA arrays for important cellular response factors. In Aim 3, any disease-toxicant interactions identified in Aim 1 and Aim 2 will be further tested in hiPSC-derived cells from HD patients with a range of different pathological repeat lengths. Results will also be analyzed by multivariate ANCOVA to determine if any of the phenotypes are repeat length dependent. This proposal will identify types of cellular stress and potential toxicants that could alter the progression of Huntington's disease and investigate the manner by which mutant Huntingtin causes this susceptibility. The proposed research will seek to identify patient-specific risk for these toxicants potentially providing individualized environmental health advice for patients (i.e. personalized environmental medicine). This funding will also contribute to my individual development as an independent researcher by allowing me to pursue these research aims and present the resulting data to the broader scientific community. As the principal investigator, I will have increased ownership of this research project and the freedom to take full advantage of my graduate training by reducing potential financial constraints.

描述（由申请人提供）：环境因子被认为与个体的遗传组成相互作用，以影响或引起神经退行性疾病。亨廷顿氏病（HD）是一种使人衰弱的神经系统疾病，具有显性单基因遗传模式。尽管如此，环境因素被认为强烈影响发病年龄和进展。我们假设环境导致细胞应激，HD脑中的神经元易受氧化应激、线粒体功能障碍和钙调节异常的影响。我们假设突变亨廷顿蛋白影响细胞应激反应，产生这种疾病特异性的脆弱性。我们将通过将细胞暴露于已知会引起相关细胞应激途径的毒物并测量毒理学表型来检验这一假设。在这里，我们建议使用诱导多能干细胞分化成纹状体神经祖细胞从患者与亨廷顿的疾病和控制作为我们的模型系统。这些细胞系含有具有完整启动子区的突变亨廷顿基因以及神经细胞类型中的患者特异性遗传背景。在具体目标1中，我们将优化产生DLX 2表达神经祖细胞的技术。然后，我们将来自两名青少年HD受试者和两名对照受试者的hiPSC的表达DLX 2的培养物暴露于细胞应激模型毒物，并评价差异活力。在《特定目标2》中，这些细胞 将用于测定毒物暴露对活性氧产生、线粒体膜电位、ATP含量和细胞溶质钙浓度的影响。将通过疾病-毒物和/或患者-毒物相互作用的多变量ANCOVA统计建模分析目标1和目标2中生成的数据。对于毒物引起的统计学显着的疾病-毒物相互作用，我们将进行蛋白质和mRNA阵列的重要细胞反应因子。在目标3中，将在来自HD患者的具有一系列不同病理重复长度的hiPSC衍生细胞中进一步测试目标1和目标2中确定的任何疾病-毒物相互作用。还将通过多变量ANCOVA分析结果，以确定任何表型是否具有重复长度依赖性。这项提案将确定细胞应激的类型和可能改变亨廷顿病进展的潜在毒物 并研究突变亨廷顿蛋白引起这种易感性的方式。拟议的研究将寻求确定这些毒物的患者特定风险，可能为患者提供个性化的环境健康建议（即个性化环境医学）。这笔资金也将有助于我作为一名独立研究人员的个人发展，使我能够追求这些研究目标，并向更广泛的科学界展示结果数据。作为首席研究员，我将增加对这个研究项目的所有权，并通过减少潜在的财务限制，充分利用我的研究生培训的自由。