Leber Hereditary Optic Neuropathy: Gene Therapy Trial

莱伯遗传性视神经病：基因治疗试验

基本信息

批准号：
8089419
负责人：
John Guy
金额：
$ 108.48万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-01 至 2013-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8089419
关键词：
ATP Synthesis Pathway Affect Amino Acids Animal Model Antibodies Apoptosis Arginine Axon Behavior Biodistribution Biological Biological Assay Blindness Cells Childhood Clinical Clinical Protocols Clinical Trials Clinical Trials Design Codon Nucleotides Collaborations Complex Consensus Cultured Cells DNA Data Development Disease Dose Effectiveness Feedback Funding Gene Delivery Genes Goals Grant Hearing Heart Histidine Human Individual Injection of therapeutic agent Investigational Drugs Investigational New Drug Application Knowledge Leber&apos s Hereditary Optic Neuropathy Mediating Mitochondria Mitochondrial DNA Mitochondrial Diseases Monoclonal Antibody R24 Mus Mutate Mutation Nature Neuro-Ocular System Nuclear Optic Disk Optic Nerve Outcome Outcome Measure Oxidative Phosphorylation Pathogenesis Patients Persons Phase Preclinical Testing Primates Principal Investigator Production Proteins Reactive Oxygen Species Reading Frames Recombinants Recovery Relative (related person)Research Research Personnel Respiratory physiology Retina Safety Schedule Series Simplexvirus Site-Directed Mutagenesis Swelling Techniques Technology Testing TimeLine Toxic effect Toxicology United States National Institutes of Health Validation Vision Visual system structure Work adeno-associated viral vector design emerging adult ganglion cell gene complementation gene therapy human disease in vivo man meetings mutant optic nerve disorder organizational structure preclinical study preclinical toxicity prevent programs research clinical testing symposium therapeutic evaluation vector

项目摘要

DESCRIPTION (provided by applicant): Our understanding of human diseases caused by mutated mitochondrial DNA has recently evolved dramatically. This is a group of untreatable disorders affecting the eye, nervous system and heart, some that have been clinically characterized over a century ago, but they are now known to be a spectrum of molecularly defined diseases. We have chosen to start with one of the most severe, the G11778A mutation in mitochondrial DNA responsible for Leber Hereditary Optic Neuropathy (LHON) a disease group renowned for causing blindness in later childhood and early adulthood. It shows little or no propensity for recovery. In previous work we have made major strides towards determining the pathogenesis and testing a treatment for LHON. First, we discovered that ND4 mutant cells have a severe reduction in ATP synthesis, even though mild reductions in complex I activity appear insufficient to induce disease. Since no technology existed to introduce DNA directly into mitochondria, we overcame this deficiency in oxidative phosphorylation by constructing a "nuclear version" of the mitochondrial gene then targeted the cytoplasmically synthesized protein to the mitochondria by using a targeting sequence appended to the reading frame (allotopic expression). When delivered to cultured cells containing a mutant ND4, respiratory function was restored. Next, we created a bona fide animal model for LHON. Using site directed mutagenesis of the nuclear version of ND4 we replaced the codon for arginine to that for histidine at amino acid 340. Injection of this construct into the mouse visual system disrupted mitochondrial cytoarchitecture, elevated reactive oxygen species, induced swelling of the optic nerve head and induced apoptosis, with a progressive demise of ganglion cells in the retina and their axons comprising the optic nerve. In contrast, ocular expression of the wild-type human ND4 subunit appeared safe, suggesting that it may be useful for treatment of patients with LHON. Here by using the scientific and clinical knowledge acquired to date we will begin the journey towards genetic therapy for human optic neuropathy and mitochondrial disease. Our goal in this application is to test the relevance of allotopic rescue of our LHON animal model; the safety of AAV mediated delivery of the ND4 gene and the effectiveness of this therapy in preventing and restoring visual loss in our patients with LHON.

描述（由申请人提供）：我们对突变的线粒体DNA引起的人类疾病的理解最近发生了巨大进化。这是一组影响眼睛，神经系统和心脏的不可治疗的疾病，有些在一个世纪前临床表征，但现在已知它们是一系列分子定义的疾病。我们选择从线粒体DNA中最严重，最严重的G11778A突变之一开始，负责Leber遗传性视神经病神经病（LHON）一个因在童年和成年初引起失明而闻名的疾病组。它几乎没有恢复的倾向。在先前的工作中，我们已经在确定发病机理和测试LHON治疗方面取得了长足的进步。首先，我们发现ND4突变细胞的ATP合成严重降低，即使复杂I活性的轻度降低似乎不足以诱导疾病。由于没有直接将DNA直接引入线粒体的技术，因此我们通过构建线粒体基因的“核版本”来克服这种缺陷，然后将细胞胞质合成的蛋白质靶向到线粒体上，通过使用靶向序列应用于阅读框架（同倍型表达）。当传递到含有突变体ND4的培养细胞时，恢复了呼吸功能。接下来，我们为Lhon创建了一个真正的动物模型。使用位点的ND4核版本的诱变，我们将精氨酸的密码子替换为氨基酸340的组氨酸的密码子。将这种构建体注射到小鼠视觉系统中破坏了线粒体细胞体系结构，反应性氧气升高，使反应性氧气升高，引起了诱导的降解，并诱发了驱动的细胞，并导致了驱逐出渐进的eNTIS notivion Demivion Demivion Demistir notivion DemiS intir intir intir intir intir intir intirip notive notive demistion demivion demist in gromistion demistion demistiv神经。相反，野生型人ND4亚基的眼部表达似乎是安全的，这表明它可能对治疗LHON患者有用。在这里，通过使用迄今为止获得的科学和临床知识，我们将开始寻求人类视神经病和线粒体疾病的基因治疗。我们在此应用中的目标是测试同种异体拯救我们LHON动物模型的相关性； AAV介导的ND4基因的递送的安全性以及该疗法在防止和恢复LHON患者视觉丧失方面的有效性。