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（由申请人提供）：拟议的奖学金项目将侧重于优化体外模型系统来测试新的亨廷顿病（HD）疗法，创建一个基因修饰策略库，旨在特异性沉默或纠正亨廷顿病中的突变基因，以及通过验证一种新的细胞模型，可以通过经济有效、稳健且可靠的方式测量 HD 相关功能障碍，从而实现这三个具体目标中的第一个目标。这将通过确认从HD患者分离的成纤维细胞培养物中的突变等位基因依赖性缺陷来实现，这将通过首先确定可以开发基因修饰策略的突变等位基因内的特定靶位点来实现。可以通过识别超过 40% 的亨廷顿舞蹈症群体中存在的突变等位基因中的单核苷酸多态性，并创建一个转录激活因子样效应子 (TALE) 文库来完成，该文库可以识别和沉默或通过针对突变等位基因中存在的单核苷酸多态性，在未来的研究中将有可能从我们的文库中专门选择患者特异性的TALE，该TALE将独立或协同地抑制该个体中的突变等位基因。适合该人基因组的 TALE 组合将有助于进一步了解 HD 患者的个性化治疗，并将有助于传播亨廷顿基因在患者特异性人类细胞系中的功能。 TALE 的创建理论上涵盖了 40% 的 HD 人群和这项探索性研究的知识可用于为绝大多数 HD 患者制定个性化治疗方案。国家神经疾病和中风研究所的使命，因为它将进一步了解亨廷顿基因的功能、蛋白质聚集和错误折叠、线粒体功能障碍以突变等位基因长度依赖性方式发生的程度以及这些缺陷的能力将通过基因改造进行纠正。在两名高级主要研究人员的专家培训下，他们在基因组靶向、基因治疗和临床试验治疗干预措施的开发方面拥有丰富的知识。这项工作将在申办者和共同申办者的直接监督下，在两个实验室之间合作进行。该奖学金将通过仔细检查患有遗传性疾病的个体的患者特异性基因修饰物，极大地增强个性化治疗的知识基础。