Novel epigenetic targets for silencing the Huntingtons disease mutation

沉默亨廷顿病突变的新表观遗传靶点

基本信息

批准号：
8960294
负责人：
Kyle Fink
金额：
$ 5.24万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-15 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8960294
关键词：
Alleles Basic Science Biological Assay Biological Models Boxing Brain California Cell Line Cell model Cells Chromosomal Stability Clinical Clinical Trials Collaborations DNA Restriction Enzymes DNA Sequence Discrimination Energy Transfer Environment Epigenetic Process Fellowship Fibroblasts Fluorescence Functional disorder Funding Future Gene Expression Gene Frequency Gene Mutation Gene Proteins Gene Targeting Gene-Modified Genes Genome Genotype Goals Hereditary Disease Human Human Cell Line Huntington Disease In Vitro Individual Knowledge Laboratories Length Libraries Life Location Measures Medical Mentors Methodology Minor Mission Mutation National Institute of Neurological Disorders and Stroke Neurodegenerative Disorders Nonhomologous DNA End Joining Pathway interactions Patients Persons Population Promoter Regions Proteins Reactive Oxygen Species Research Research Personnel Research Training Silene Single Nucleotide Polymorphism Site Supervision System Technology Testing Therapeutic Therapeutic Intervention Time Training Transcription Coactivator Transcription Repressor/Corepressor Translations Universities Work career development cost effective design gene therapy human Huntingtin protein in vitro Model knowledge base mitochondrial dysfunction multidisciplinary mutant nervous system disorder novel nuclease personalized medicine personalized therapeutic protein aggregation protein misfolding public health relevance repository therapy development vector

项目摘要

DESCRIPTION (provided by applicant): The proposed fellowship project will focus on optimizing an in vitro model system to test novel Huntington's disease (HD) therapeutics, creating a library of gene modification strategies aimed at silencing or correcting specifically th mutant gene in HD, and demonstrating robust and durable knockdown of HD-related cellular deficits using gene modification. The first of these three specific aims will be accomplished by validating a novel cellular model in which HD related dysfunction can be measured in a cost-effective, robust, and reproducible manner. This will be done by confirming mutant-allele-dependent deficits in fibroblast cultures isolated from HD patients. The second goal will be accomplished by first identifying specific target sites within the mutant allele for which gene modifying strategies can be developed. This will be done by identifying single nucleotide polymorphisms in the mutant allele that exist in greater than 40% of the Huntington's population and creating a library of transcription activator-like effectors (TALE) that can identify and silene or correct the mutant gene. By targeting single nucleotide polymorphisms that exist in the mutant allele, it will be possible in future studies to choose a patient-specific TALE from our library that will specifically suppress the mutant allele in that individual either independently o acting synergistically with a combination of TALEs suited for that persons genome. The results from this proposal will help further the knowledge of personalized treatment for patients suffering from HD and will help disseminate the function of the huntingtin gene in patient-specific human cell lines. A working library of TALE will be created that theoretically covers over 40% of the HD population and knowledge from this exploratory study can be applied to create personalized therapeutics for the vast majority of HD patients. This fellowship is in line with the mission of the National Institute of Neurological Disorders and Stroke as it will further the knowledge of the function of huntingtin gene, protein aggregation and misfolding, and the extent to which mitochondrial dysfunction occurs in a mutant allele- length dependent manner and the ability of these deficits to be corrected with gene modification. This fellowship will be conducted under the expert training of two senior principle investigators with extensive knowledge of genome targeting, gene therapy, and the development of therapeutic interventions for clinical trials. The work will be performed in collaboration between the two labs with direct supervision from the sponsor and co-sponsor. The fellowship will greatly enhance the knowledge base for personalized therapies, through careful examination of patient- specific gene modifiers for individuals suffering from genetic disorders.

描述（由适用提供）：拟议的奖学金项目将着重于优化一种测试新型亨廷顿疾病（HD）疗法的体外模型系统，创建了旨在在HD中专门纠正突变基因的基因修饰策略库，并使用基因定义的HD细胞定义来证明可靠且耐用的敲低敲低敲低的基因。这三个特定目的中的第一个将通过验证一个新型的细胞模型来实现，在该模型中可以以成本效益，健壮和可再现的方式测量HD相关的功能障碍。这将通过确认依赖突变体 - 依赖性的依赖性的成纤维细胞培养物中的成纤维细胞培养物来完成。第二个目标将通过首先确定可以开发基因修改策略的突变等位基因中的特定目标位点来实现。这将通过鉴定突变等位基因中的单核苷酸多态性来完成，该多态性大于亨廷顿人口的40％以上，并创建一个可以识别和硅的转录激活效应（Tale）库（Tale）库。通过靶向突变等位基因中存在的单个核丁物多态性，在以后的研究中，可以从我们的图书馆中选择一个特定于患者的故事，该故事将专门抑制突变等位基因在该个体中独立地与适合该人的故事组合共同行动的人。该提案的结果将有助于进一步了解患有HD患者的个性化治疗，并有助于传播亨廷顿基因在患者特异性人细胞系中的功能。将创建一个工作的故事库，理论上涵盖这项探索性研究的高清人群中有40％可以用于为绝大多数HD患者创建个性化疗法。该奖学金与国家神经系统疾病和中风研究所的使命将进一步了解亨廷汀基因的功能，蛋白质聚集和错误折叠，以及线粒体功能障碍的程度，以突变等位基因等位基因的依赖性方式以及这些定义与基因修饰校正的能力发生。该奖学金将进行在两名高级原则研究者的专家培训下，对基因组靶向，基因治疗以及临床试验治疗干预理论的发展有了广泛的了解。这项工作将在两个实验室之间的合作下，并在赞助商和共同赞助商的直接监督下进行。通过仔细检查患有遗传疾病的个体的患者特定基因修饰剂，该研究金将大大增强个性化疗法的知识库。