Rare Variant Whole Genome Analysis and iPSC Validation of Putative Genetic Modifiers of Huntington Disease

亨廷顿病的假定遗传修饰物的罕见变异全基因组分析和 iPSC 验证

基本信息

批准号：
9925102
负责人：
STEVEN M FINKBEINER
金额：
$ 64.38万
依托单位：
J. DAVID GLADSTONE INSTITUTES
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-04-01 至 2021-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9925102
关键词：
Affect Age Age of Onset Alzheimer&apos s Disease Appearance Autophagocytosis Biological CRISPR/Cas technology Cells Cellular Stress Clinical Clinical Trials Corpus striatum structure Data Data Set Development Disease Disease Progression Enrollment Exhibits Family Family member Funding Gene Expression Gene Proteins Generations Genes Genetic Genetic Diseases Genetic Polymorphism Genome Goals Grant Heritability Human Huntington Disease Huntington gene Huntington protein Impairment Individual Institutes Knowledge Label Late-Onset Disorder Late-Onset Huntington Disease Length Link Measures Medical Records Metabolic Clearance Rate Methods Mind Modeling Molecular Target Monitor Nature Nerve Degeneration Neurites Neurodegenerative Disorders Neurons Onset of illness Optics Pathway interactions Patients Pattern Pharmaceutical Preparations Phenotype Physiologic pulse Population Process Property Proteins Recording of previous events Research Role Statistical Methods Survival Analysis Symptoms System Systems Biology Technology Testing Therapeutic Time Toxic effect Translating United States National Institutes of Health Validation Variant base cellular imaging clinical phenotype design disease phenotype gene product genetic linkage analysis genetic variant genome analysis genome sequencing genome wide association study improved induced pluripotent stem cell insight member multicatalytic endopeptidase complex mutant new technology new therapeutic target novel novel therapeutics overexpression programs rare variant robotic microscopy segregation selective expression stem cell model whole genome

项目摘要

PROJECT SUMMARY The goal of our studies is to identify and validate genes, proteins, and biological pathways that modulate neurodegeneration induced by mutant huntingtin (mHtt), the protein that causes Huntington's disease (HD). This knowledge will provide new insights into the underlying mechanisms of HD and may reveal novel therapeutic targets that are more druggable than mHtt. While mHtt is the major cause for HD, a number of studies have indicated that genetic modifiers interact with mHtt to affect progression of neurodegeneration in HD. In fact, a substantial genetic contribution to HD is not accounted for solely by the gene that encodes mHtt, or by the few modifiers that have been identified by other research groups. We hypothesize that rare genetic variants contribute to the disease onset and progression of HD that have been missed by genome-wide association studies (GWAS) or candidate-based approaches. With this in mind, we conducted whole-genome sequencing (WGS) on multiple HD families and identified candidates in novel genes not previously implicated in HD. They are involved in protein clearance and other cellular pathways that may contribute to neurodegeneration in HD. We provide direct evidence, for the first time, that a subset of these candidates modify neurodegeneration of human striatal-like HD iPSC-derived neurons (HD striatal i-neuron). In the proposed studies, we will further validate and investigate the mechanisms by which these potential genetic modifiers modulate neurodegeneration and expand our analysis to additional variants and their cellular pathways that contribute to neurodegeneration in HD. Human neuron models recapitulate several key features of HD, and a form of cellular imaging called robotic microscopy (RM) enables high-throughput (HT), high-content, longitudinal single-neuron analysis of these models. The data sets generated by RM reveal different aspects of neurodegeneration, including survival, analyzed by powerful statistical methods, or changes in neurite length, which is a predictor of cellular stress. Our toolbox uses other powerful approaches to assess a candidates' effects on neurodegeneration, such as an optical-pulse labeling (OPL) technology that can measure the rate of clearance of proteins by proteasome activity or autophagy within single cells. We have an NIH X01 grant that is sequencing 104 additional members of 19 new HD families for which we have extensive medical records and clinical history on. We will extend our WGS analysis to these families and combine the data to identify a more complete set of interacting gene partners and pathways and to help focus our list of current candidates that contribute to HD onset and trajectory. New putative variants will be tested in our human HD i-neuron model to validate them as potential genetic modifiers and to better define cellular pathways involved in modulating onset of HD. The discovery of novel genetic modifiers of HD will further elucidate the disease mechanisms in HD and identify new directions for developing disease-modifying therapeutics and for stratifying HD populations for more successful clinical trials.

项目摘要我们研究的目的是识别和验证调节基因，蛋白质和生物途径由突变体亨廷顿蛋白（MHTT）诱导的神经变性，这是导致亨廷顿氏病（HD）的蛋白质。这知识将为HD的潜在机制提供新的见解，并可能揭示新的治疗性比MHTT更毒的目标。虽然MHTT是HD的主要原因，但许多研究已表明遗传修饰符与MHTT相互作用，以影响HD中神经退行性的进展。实际上，编码MHTT的基因或少数几个基因不考虑对HD的实质性遗传贡献由其他研究小组确定的修饰符。我们假设稀有遗传变异有助于疾病的发作和HD的发展，而全基因组关联却遗漏了研究（GWAS）或基于候选的方法。考虑到这一点，我们进行了全基因组测序（WG）在多个HD家族上，并在以前与HD相关的新基因中鉴定了候选者。他们参与蛋白质清除率和其他可能导致HD神经退行性的细胞途径。我们首次提供了直接证据，表明这些候选者的子集修改了神经退行性的。人纹状体样HD IPSC衍生的神经元（HD纹状体I-Neuron）。在拟议的研究中，我们将进一步验证和研究这些潜力的机制遗传修饰符调节神经变性并将我们的分析扩展到其他变体及其细胞导致HD神经退行性的途径。人类神经元模型概括了几个关键特征高清和一种称为机器人显微镜（RM）的细胞成像形式可实现高通量（HT），高含量，这些模型的纵向单神经元分析。 RM生成的数据集揭示了神经变性，包括生存，通过强大的统计方法分析或神经突长度的变化，这是细胞应激的预测指标。我们的工具箱使用其他强大的方法来评估候选人的效果关于神经变性的，例如光脉冲标签（OPL）技术，可以测量通过蛋白酶体活性或单细胞内自噬清除蛋白质。我们有NIH X01赠款测序我们有大量医疗记录的19个新高清家庭中的另外104位成员临床病史。我们将将WGS分析扩展到这些家庭，并结合数据以识别更多完整的互动基因伙伴和途径集，并帮助我们集中我们当前的候选人名单有助于高清发作和轨迹。新的推定变体将在我们的人类高清I-Neuron模型中进行测试将它们验证为潜在的遗传修饰符，并更好地定义参与调节发作的细胞途径高清。新型HD遗传修饰剂的发现将进一步阐明HD和确定开发疾病改良治疗剂和分层高清种群的新方向成功的临床试验。