Mito-Targeted AAV to treat Leber Hereditary Optic Neuropathy caused by ND4 mutations

Mito 靶向 AAV 治疗 ND4 突变引起的 Leber 遗传性视神经病

基本信息

批准号：
10554399
负责人：
Byron L Lam
金额：
--
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：
2019
资助国家：
美国
起止时间：
2019-03-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10554399
关键词：
ATP Synthesis Pathway Acute Adverse effects Affect Amino Acids Animal Model Arginine Atrophic Axon Bilateral Biological Birth Blindness Capsid Categories Cell model Cells Characteristics Chronic Clinical Clinical Trials Clinical Trials Design Codon Nucleotides Complex Cytochrome c Reductase DNA DNA Maintenance DNA biosynthesis DNA delivery Data Dependovirus Development Disease Effectiveness FDA approved Family Family member Funding Future Gene Delivery Gene Transfer Generations Genes Genetic Code Glaucoma Goals Histidine Histopathology Human Human Characteristics Induction of Apoptosis Infrastructure Injections Investigational Drugs Investigational New Drug Application Lead Leber&apos s Hereditary Optic Neuropathy Leber&apos s amaurosis Macaca mulatta Microinjections Mitochondria Mitochondrial DNA Mitochondrial Diseases Modeling Mus Mutate Mutation NADH dehydrogenase (ubiquinone)Natural History Neurodegenerative Disorders Nuclear Optic Disk Optic Nerve Organelles Oxidative Phosphorylation Pathogenesis Patients Phase I Clinical Trials Phase I/II Clinical Trial Phase I/II Trial Phenotype Preclinical Testing Primates R24 Rare Diseases Reactive Oxygen Species Regulatory Element Research Research Personnel Respiration Respiratory Chain Respiratory physiology Rest Retina Retinal Diseases Rodent Safety Series Site-Directed Mutagenesis Structure Swelling Technology Testing Time Tissues Toxicology Transgenic Mice Transgenic Organisms Translational Research Treatment Efficacy United States National Institutes of Health Viral Visual System Work adeno-associated viral vector antibody test blastocyst clinically relevant design effective therapy efficacy testing experience ganglion cell gene therapy in vivo intravitreal injection mouse model multidisciplinary mutant nonhuman primate optic nerve disorder pre-Investigational New Drug meeting preclinical safety prevent promoter prospective retinal ganglion cell degeneration safety study safety testing sight restoration translational study vector vector biodistribution visual optics

项目摘要

Research Summary Mutations in mitochondrial DNA (mtDNA) lead to a spectrum of neurodegenerative diseases for which no effective treatment exists. The most common of these is Leber hereditary optic neuropathy (LHON) caused by mutations in NADH dehydrogenase subunit genes (ND1, ND4 or ND6), which is complex I of the respiratory chain. Therapies for LHON in common with all disorders caused by mutated mtDNA are inadequate, in large part because of the barrier in delivering DNA into the organelle. We have successfully broken through this barrier by developing a pioneering adeno-associated virus (AAV) vector to which a mitochondrial targeting sequence (MTS) was appended to the viral capsid. The modified vector delivered the ND4 gene directly to the mitochondria for reversal of visual loss in mice with mutated G11778A ND4 responsible for more than half of all LHON cases, the rest caused by mutated ND1 and ND6. We will now design, modify and test the efficacy and safety of a clinically relevant vector for treatment of this mitochondrial disease by delivery of genes encoding the promoter and regulatory elements of a normal mitochondrially encoded human ND4 subunit to affected cells and tissues for rescue of cultured human LHON cells harboring the NADH dehydrogenase subunit 4 mutation causing LHON and then transgenic mice and primates with mutated ND4. The ND4 mouse we developed by injection of a MTS AAV containing mutated G11778A ND4 into the rodent blastocyst has visual loss progressing to blindness a year after birth, optic nerve head swelling followed by atrophy and degeneration of retinal ganglion cells, which are the characteristic hallmarks of LHON patients. Our goals are: (A) To facilitate translational studies for LHON by developing a clinical grade MTS AAV vector that accommodates the wild-type ND4 LHON gene and regulatory elements in a single AAV cassette. (B) To test expression to rescue respiration in cybrid cells and the visual system of mice and primates with mutated G11778A ND4. (C) To evaluate biological effects of intravitreal delivery of MTS AAV vectors in normal rodents and primates that result in mitochondrial gene transfer without adverse effects. (D) To develop an IND application for a single AAV containing normal ND4 for a future phase I/II clinical trial designed to restore the vision of patients with the commonest and most severe LHON mutation and prevent visual loss in their family members in a U10 application to follow successful completion of this R24.

研究摘要线粒体DNA（mtDNA）中的突变导致一系列神经退行性疾病的光谱存在有效的治疗。其中最常见的是由 NADH脱氢酶亚基基因（ND1，ND4或ND6）中的突变，这是呼吸的复杂I 链。与突变mtDNA引起的所有疾病共同的LHON疗法不充分部分是因为将DNA输送到细胞器中的障碍。我们已经成功解决了这个通过开发与线粒体靶向的开创性腺相关病毒（AAV）媒介的障碍将序列（MT）添加到病毒式衣壳上。修改的向量将ND4基因直接传递到突变的G11778A ND4的小鼠视觉丧失的线粒体逆转负责超过一半 LHON病例，其余由突变的ND1和ND6引起。现在，我们将设计，修改和测试功效和通过递送编码基因的临床相关载体治疗这种线粒体疾病的安全性正常线粒体编码的人ND4亚基的启动子和调节元素，用于受影响的细胞和用于营救含有NADH脱氢酶亚基4突变的培养的人类LHON细胞的组织引起LHON，然后引起转基因小鼠和突变的ND4灵长类动物。我们开发的ND4鼠标将含有突变的G11778A ND4的MTS AAV注入啮齿动物胚泡中出生一年后失明，视神经头肿胀，随后萎缩和视网膜变性神经节细胞，这是LHON患者的特征标志。我们的目标是：（a）促进 LHON的翻译研究通过开发适合野生型的临床级MTS AAV载体单个AAV盒中的ND4 LHON基因和调节元素。（b）测试表达以挽救呼吸在Cybrid细胞和小鼠的视觉系统中，具有突变的G11778A ND4的灵长类动物。（c）评估在正常啮齿动物和灵长类动物中玻璃体内递送MTS AAV载体的生物学作用，从而导致线粒体基因转移而没有不利影响。（d）为单个AAV开发IND应用程序在未来的I/II期临床试验中包含正常的ND4，旨在恢复患者的视力最常见，最严重的LHON突变，并防止其U10中的家人视觉丧失遵循此R24成功完成的申请。