Novel epigenetic targets for silencing the Huntingtons disease mutation

 描述(由申请人提供)：拟议的奖学金项目将专注于优化体外模型系统以测试新的亨廷顿病(HD)疗法，创建一个旨在沉默或纠正HD中特定突变基因的基因修改策略库，并展示通过基因修改强大而持久地击倒HD相关细胞缺陷。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，HD相关功能障碍可以以经济高效、健壮和可重复的方式进行测量。这将通过确认从HD患者分离的成纤维细胞培养中的突变-等位基因依赖缺陷来完成。第二个目标将通过首先确定突变等位基因中可以开发基因修改策略的特定靶点来实现。这将通过识别存在于超过40%的亨廷顿人群中的突变等位基因中的单核苷酸多态性，并创建一个转录激活器样效应器(TALL)库来完成，该库可以识别和沉默或纠正突变基因。通过针对突变等位基因中存在的单核苷酸多态，在未来的研究中，有可能从我们的文库中选择一个特定于患者的故事，它将独立地或与适合于该人基因组的故事的组合协同作用，特异性地抑制该个体中的突变等位基因。这项建议的结果将有助于进一步了解HD患者的个性化治疗，并将有助于在特定患者的人类细胞系中传播Huntingtin基因的功能。将创建一个工作的故事库，理论上涵盖 40%的HD人群和这项探索性研究的知识可以应用于为绝大多数HD患者创造个性化的治疗方法。这项奖学金符合 国家神经疾病和中风研究所的任务，因为它将进一步了解Huntingtin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的等位基因长度依赖的方式发生的程度以及这些缺陷通过基因修改得到纠正的能力。本次联谊会将进行 在两名资深首席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具有丰富的知识。这项工作将在赞助商和共同赞助商的直接监督下，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疾病的患者特定的基因修饰物，极大地增强个性化治疗的知识库。

 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.