Modification of AdenoAssociated Virus to Deliver DNA Directly to Mitochondria

修饰腺相关病毒以将 DNA 直接递送至线粒体

基本信息

批准号：
9767197
负责人：
Hong Yu
金额：
$ 36.81万
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：
2007
资助国家：
美国
起止时间：
2007-09-01 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9767197
关键词：
ATP Synthesis Pathway Adverse effects Affect Alleles American Animals Atrophic Autopsy Biological Birth Blindness Capsid Proteins Categories Cell Line Cell physiology Cells Characteristics Childhood Clinical Clinical Trials Complex Cytochrome c Reductase DNA DNA delivery Defect Dependovirus Development Disease Electroretinography Epitopes Eye Gene Delivery Gene Expression Gene Transfer Genes Genetic Code Goals Grant Heart Homologous Gene Human Injections Knowledge Laser Scanning Confocal Microscopy Lead Leber&apos s Hereditary Optic Neuropathy Length Link Mitochondria Mitochondrial DNA Mitochondrial Diseases Modification Molecular Mus Mutate Mutation NADH dehydrogenase (ubiquinone)Nerve Degeneration Neuro-Ocular System Neurodegenerative Disorders Noise Nucleotides Optic Atrophy Optic Disk Optic Nerve Organelles Oxidative Phosphorylation Pathogenesis Patients Pattern Phase Preclinical Testing Prevention Proteins Rare Diseases Recovery Respiration Respiratory Chain Retina Rodent Safety Swelling Technology Testing Tissues Toxic effect Transgenic Mice Transgenic Organisms Treatment Efficacy Vertebral column Viral Virion Virus Vision Visual Work adeno-associated viral vector blastocyst clinically relevant design effective therapy efficacy testing emerging adult experimental study ganglion cell gene therapy human disease mutant new technology non-invasive imaging optic nerve disorder prevent promoter success transgene expression translational study vector visual optics

项目摘要

Mutations in mitochondrial DNA lead to a spectrum of neurodegenerative diseases for which no effective treatment exists. The most common of these is Leber's hereditary optic neuropathy (LHON) caused by mutations in subunit genes (ND1, ND4 or ND6) of NADH dehydrogenase, which is complex I of the respiratory chain. This application builds on the successes of the current grant that pioneered an adeno-associated virus (AAV) vector to which a mitochondrial targeting sequence (MTS) was appended to the viral envelope. The modified vector delivered the NADH dehydrogenase subunit 4 (ND4) gene directly to the mitochondria for prevention of visual loss in mice also injected with a mutated G11778A ND4 responsible for half of all LHON cases. 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 normal human ND1, ND4 and ND6 subunits to affected cells and tissues; for rescue of cultured human LHON cells harboring each of the three mutations causing LHON and also a transgenic mouse we developed by injection of the MTS AAV containing mutant human ND4 into the blastocyst. This mouse has visual loss progressing to blindness a year after birth, optic nerve head swelling followed by atrophy and degeneration of ganglion cells, which are the characteristic hallmarks of LHON patients. Our Aims are: (1) To facilitate translational studies for LHON by developing MTS AAV vectors to deliver each of the ND1, ND4 and ND6 subunit genes directly to the mitochondria and test expression to rescue respiration in cybrid cells with100% mutated ND1, ND4 or ND6. We also develop a single AAV cassette that accommodates all three genes that would require a single IND for FDA approval. (2) To evaluate biological effects of intravitreal delivery of MTS AAV vectors in normal mice that result in mitochondrial gene transfer without adverse effects.(3) To rescue visual loss and optic nerve degeneration in transgenic LHON mice, we carry out treatment at stages that closely parallel the human disease before and after RGC loss and optic atrophy, using viral titers in LHON transgenic mice that in our current experiments resulted in rescue without adverse effects. We hope to identify the conditions for long-term rescue of optic neuropathy in mice, so that this approach can be tested in a phase I/II clinical designed to restore the vision of our patients with all three of the common LHON mutations.

线粒体 DNA 突变会导致一系列神经退行性疾病，而这些疾病目前尚无定论。存在有效的治疗方法。其中最常见的是莱伯遗传性视神经病 (LHON) 由 NADH 脱氢酶亚基基因（ND1、ND4 或 ND6）突变引起，这是呼吸链的复合体 I。该应用程序建立在目前的资助开创了腺相关病毒（AAV）载体，其中线粒体靶向序列（MTS）被附加到病毒包膜上。修改后的载体交付将 NADH 脱氢酶亚基 4 (ND4) 基因直接连接至线粒体以预防注射了突变 G11778A ND4 的小鼠视力丧失占所有 LHON 的一半案例。我们现在将设计、修改和测试临床相关载体的功效和安全性通过传递编码正常人类线粒体的基因来治疗这种线粒体疾病受影响细胞和组织的 ND1、ND4 和 ND6 亚基；拯救培养人类LHON 携带导致 LHON 的三种突变的细胞以及我们的转基因小鼠通过将含有突变型人 ND4 的 MTS AAV 注射到囊胚中而开发。这小鼠出生一年后视力丧失并逐渐失明，视神经乳头肿胀其次是神经节细胞的萎缩和变性，这是神经节细胞的特征性标志 LHON 患者。我们的目标是： (1) 通过开发促进 LHON 的转化研究 MTS AAV 载体将 ND1、ND4 和 ND6 亚基基因直接递送至线粒体并测试表达以挽救具有 100% 突变 ND1 的 cybrid 细胞的呼吸， ND4 或 ND6。我们还开发了一个 AAV 盒，可容纳所有三个基因需要单一 IND 才能获得 FDA 批准。 (2) 评价玻璃体内注射的生物学效应将 MTS AAV 载体递送至正常小鼠体内，无需进行线粒体基因转移 (3) 挽救转基因LHON的视力丧失和视神经变性小鼠，我们在与人类疾病前后非常相似的阶段进行治疗 RGC 损失和视神经萎缩，使用 LHON 转基因小鼠中的病毒滴度，在我们目前的研究中实验结果是救援没有产生不良影响。我们希望确定条件对小鼠视神经病变的长期挽救，以便可以在 I/II 期测试该方法临床旨在恢复患有所有三种常见 LHON 的患者的视力突变。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Comparison of different gene-therapy methods to treat Leber hereditary optic neuropathy in a mouse model.

比较不同基因治疗方法在小鼠模型中治疗莱伯遗传性视神经病变的方法。

DOI：
发表时间：
2023
期刊：
影响因子：
0
作者：
Velmurugan, Sindhu;Chou, Tsung;Eastwood, Jeremy D;Porciatti, Vittorio;Liu, Yuan;Hauswirth, William W;Guy, John;Yu, Hong
通讯作者：
Yu, Hong