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

• 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
• 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

 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，以测试新型亨廷顿病（HD）治疗方法，创建基因修饰策略库，旨在沉默或纠正HD中的特定突变基因，并证明使用基因修饰对HD相关细胞缺陷的稳健和持久敲除。这三个具体目标中的第一个将通过验证一种新的细胞模型来实现，在该模型中，可以以具有成本效益的、稳健的和可再现的方式测量HD相关功能障碍。这将通过确认从HD患者中分离的成纤维细胞培养物中的mu等位基因依赖性缺陷来完成。第二个目标将通过首先鉴定突变等位基因内的特异性靶位点来实现，针对所述特异性靶位点可以开发基因修饰策略。这将通过鉴定存在于超过40%的亨廷顿人群中的突变等位基因中的单核苷酸多态性并创建可以鉴定和沉默或校正突变基因的转录激活因子样效应物（TALE）文库来完成。通过靶向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选择患者特异性TALE，其将特异性抑制该个体中的突变等位基因，或者独立地或者与适合于该人基因组的TALE组合协同作用。该提案的结果将有助于进一步了解患有HD的患者的个性化治疗，并将有助于在患者特异性人类细胞系中传播亨廷顿基因的功能。将创建一个TALE工作库，理论上涵盖 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患者创建个性化治疗。该奖学金符合 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，以及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，以及这些缺陷通过基因修饰得到纠正的能力。该奖学金将在 在两名资深研究人员的专家培训下，他们拥有基因组靶向、基因治疗和临床试验治疗干预措施开发的广泛知识。在申办方和共同申办方的直接监督下，两个实验室将合作开展这项工作。该奖学金将通过仔细检查患有遗传疾病的个人的患者特异性基因修饰剂，大大增强个性化治疗的知识基础。