Targeting mitochondrial gene expression in the retina

靶向视网膜中的线粒体基因表达

基本信息

批准号：
6985535
负责人：
Alfred S Lewin
金额：
$ 35.92万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-01 至 2010-08-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Maternally inherited diseases associated with mutations in mitochondrial DNA are generally multisystemic, typically affecting the central nervous system, heart, skeletal muscles and endocrine glands. It has been difficult to understand the pathogenic processes associated with such disorders, much less to develop treatments, because of the lack of animal models that recapitulate the characteristics of mitochondrial disease. In recent years, we have succeeded in using the tools of gene therapy, namely the viral-mediated transfer of ribozymes, to create animal models with a common feature of many mitochondrial diseases - optic nerve degeneration. Ribozymes specific for a nucleus-encoded subunit (NDUFA1) of Complex I of the mitochondrial respiratory chain lead to damage similar to that seen in Leber Hereditary Optic Neuropathy, the most common inherited mitochondrial disease. We have recently demonstrated similar pathology using a ribozyme targeted to another subunit (ND4) of the same enzyme that is produced within mitochondria. In this proposal, we plan to develop mouse models that capture another mitochondrial disorder called NARP and to take the first steps toward gene therapy of these devastating diseases. We propose: (1) To develop ribozymes that can be delivered to mitochondria to create mouse models with deficits in the ATP6 subunit of the mitochondrial ATP synthase (Complex V); and (2) To treat the animal models developed by ribozyme knockdown of ATP6 and ND4 by delivering nuclear genes that replace the deficient mitochondrial gene product, a procedure termed allotopic expression. Since it has already been established that allotropic expression can rescue mitochondrial defects in cultured cells, our second aim stands independently of the first: It is important to test allotopic expression in animal tissues that are typically affected in mitochondrial disease. While we are testing specific hypotheses concerning how mitochondrial damage causes retinopathy and optic neuropathy, our ultimate aim is developing treatments - either genetic or pharmacological - for mitochondrial disease.

描述（由申请人提供）：与线粒体 DNA 突变相关的母体遗传疾病通常是多系统的，通常影响中枢神经系统、心脏、骨骼肌和内分泌腺。由于缺乏概括线粒体疾病特征的动物模型，很难理解与此类疾病相关的致病过程，更不用说开发治疗方法了。近年来，我们成功地利用基因治疗工具，即病毒介导的核酶转移，建立了具有许多线粒体疾病共同特征——视神经变性的动物模型。线粒体呼吸链复合物 I 的核编码亚基 (NDUFA1) 特异的核酶会导致类似于莱伯遗传性视神经病（最常见的遗传性线粒体疾病）中所见的损害。我们最近使用一种核酶证明了类似的病理学，该核酶靶向线粒体内产生的同一酶的另一个亚基（ND4）。在这项提案中，我们计划开发捕获另一种称为 NARP 的线粒体疾病的小鼠模型，并为这些破坏性疾病的基因治疗迈出第一步。我们建议：（1）开发可递送至线粒体的核酶，以创建线粒体 ATP 合酶（复合体 V）的 ATP6 亚基缺陷的小鼠模型； (2) 通过传递替代缺陷线粒体基因产物的核基因来治疗通过核酶敲低 ATP6 和 ND4 开发的动物模型，这一过程称为同位素表达。由于已经确定同位素表达可以挽救培养细胞中的线粒体缺陷，因此我们的第二个目标独立于第一个目标：测试通常受线粒体疾病影响的动物组织中的同位素表达非常重要。虽然我们正在测试有关线粒体损伤如何导致视网膜病变和视神经病变的具体假设，但我们的最终目标是开发针对线粒体疾病的遗传或药物治疗方法。