Using gene targeting to address the role of LRRK2 in Parkinson's disease

利用基因靶向解决 LRRK2 在帕金森病中的作用

• 批准号: 7126106
• 负责人: BRUCE S HOSTAGER
• 金额: $ 16.59万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-16 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7126106
• 关键词: Parkinson' s disease; cell line; dopamine; gene expression; gene targeting; genes; genetics; proteins; role

DESCRIPTION (provided by applicant): Each year in the United States alone there are approximately 50,000 new.cases of Parkinson's disease. Parkinson's disease (PD) results from the malfunction or death of brain cells responsible for the production of the neurotransmitter dopamine. Dopamine deficiency results in tremor and degraded motor control. The etiology of Parkinson's is complex, and in most cases probably involves a combination of environmental and genetic factors. The advent of powerful genetic techniques has allowed the rapid identification of genes potentially associated with the development of PD. However, the subsequent biochemical characterization of the proteins encoded by these genes has been a bottleneck in understanding their roles in disease. The major goal of this proposal is to develop a new approach that will facilitate the characterization of PD associated proteins. The disruption of gene expression is an extremely powerful approach for defining the physiological roles of gene products. Recently, a technique known as RNA interference has allowed suppression of gene expression in a variety of experimental systems. However, it is often not possible to entirely eliminate the expression of a protein using this approach. Alternatively, gene sequences can be disrupted directly using a process known as homologous recombination. This approach offers complete and permanent disruption of targeted genes but has worked well only in a limited number of cell lines. We have modified the approach to potentially allow its use in many additional cell lines, including cell lines useful in PD research. As proof of principle, we will disrupt the gene encoding dardarin in a neuronal cell line. Although the function of dardarin is currently unknown, mutations in the gene (LRRK2) encoding the protein are associated with approximately 5% of all familial cases of Parkinson's disease. Using the dardarin deficient cell line, we will begin to examine the role of this novel protein in the physiology of neuronal cells. The development of a homologous recombination approach amenable to neuronal cell lines will facilitate the characterization of other PD-associated proteins. Characterization of the physiological roles of known PD associated proteins will lead to the identification of the biochemical pathways susceptible to damage in dopamine-producing neurons. This knowledge will aid in the identification of the environmental triggers and additional genetic factors that participate in the development of Parkinson's disease.

描述（由申请人提供）：每年仅在美国就有大约50，000例新的帕金森病病例。帕金森病（PD）是由负责产生神经递质多巴胺的脑细胞的功能障碍或死亡引起的。多巴胺缺乏会导致震颤和运动控制能力下降。帕金森氏症的病因很复杂，在大多数情况下可能涉及环境和遗传因素的结合。强大的遗传学技术的出现使得能够快速鉴定可能与PD发展相关的基因。然而，这些基因编码的蛋白质的后续生物化学表征一直是理解其在疾病中的作用的瓶颈。该提案的主要目标是开发一种新的方法，将有助于PD相关蛋白的表征。基因表达的破坏是一个非常强大的方法来定义基因产物的生理作用。最近，一种被称为RNA干扰的技术已经允许在各种实验系统中抑制基因表达。然而，使用这种方法通常不可能完全消除蛋白质的表达。或者，可以使用称为同源重组的方法直接破坏基因序列。这种方法提供了对靶基因的完全和永久破坏，但仅在有限数量的细胞系中效果良好。我们已经修改了该方法，以潜在地允许其在许多其他细胞系中使用，包括在PD研究中有用的细胞系。作为原理的证明，我们将破坏神经元细胞系中编码达达林的基因。虽然dardarin的功能目前尚不清楚，但编码该蛋白的基因（LRRK2）突变与所有帕金森病家族病例的约5%相关。使用dardarin缺陷细胞系，我们将开始检查这种新蛋白在神经元细胞生理学中的作用。开发一种适用于神经元细胞系的同源重组方法将有助于表征其他PD相关蛋白。已知PD相关蛋白的生理作用的表征将导致识别对多巴胺产生神经元中的损伤敏感的生化途径。这些知识将有助于识别参与帕金森病发展的环境触发因素和其他遗传因素。