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

基本信息

批准号：
8501026
负责人：
Andrew M Tidball
金额：
$ 2.69万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-06-01 至 2014-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8501026
关键词：
Affect Age 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

项目摘要

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）：环境因素被认为与个体的基因构成相互作用，从而影响或引起神经退行性疾病。亨廷顿病 (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，环境因素被认为会强烈影响疾病的发病年龄和进展。我们假设环境会导致 HD 大脑中的神经元容易受到的细胞应激，例如氧化应激、线粒体功能障碍和钙失调。我们假设突变的亨廷顿蛋白会影响细胞应激反应，从而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发相关细胞应激途径的毒物并测量毒理学表型来测试这一假设。在这里，我们建议使用来自亨廷顿病患者和对照的分化为纹状体神经祖细胞的诱导多能干细胞作为我们的模型系统。这些细胞系含有突变的亨廷顿基因，具有完整的启动子区域以及神经细胞类型中患者特异性的遗传背景。在具体目标 1 中，我们将优化生成表达 DLX2 的神经祖细胞的技术。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 DLX2 的培养物暴露于细胞应激模型毒物中，并评估差异活力。在具体目标 2 中，这些相同的单元格将用于测定有毒物质暴露对活性氧产生、线粒体膜电位、ATP 含量和胞质钙浓度的影响。目标 1 和目标 2 中生成的数据将通过疾病-毒物和/或患者-毒物相互作用的多变量 ANCOVA 统计模型进行分析。对于引起统计学上显着的疾病-毒物相互作用的毒物，我们将对重要的细胞反应因子进行蛋白质和 mRNA 阵列。在目标 3 中，目标 1 和目标 2 中确定的任何疾病毒物相互作用都将在来自 HD 患者的具有一系列不同病理重复长度的 hiPSC 衍生细胞中进行进一步测试。还将通过多变量 ANCOVA 分析结果，以确定是否有任何表型依赖于重复长度。该提案将确定可能改变亨廷顿病进展的细胞应激类型和潜在毒物并研究突变亨廷顿蛋白导致这种易感性的方式。拟议的研究将寻求确定这些有毒物质的患者特定风险，从而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）。这笔资金还将有助于我作为独立研究人员的个人发展，让我能够追求这些研究目标并将结果数据呈现给更广泛的科学界。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来充分利用我的研究生培训的自由。