Novel Gene Therapy Strategies for Canavan Disease

卡纳万病的新基因治疗策略

基本信息

批准号：
10338159
负责人：
Guangping Gao
金额：
$ 42.24万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10338159
关键词：
Acetyltransferase Adverse effects Age Animals Aspartic Acid Aspartoacylase Astrocytes Behavioral Bioinformatics Brain Brain region Canavan Disease Cell Line Cells Cessation of life Chickens Childhood Clinical DNA cassette Data Defect Disease Dose Elements Energy Metabolism Engineering Ensure Epigenetic Process Etiology Functional disorder Funding Genes Goals Head Homeostasis Human Impaired cognition Impairment In Vitro Investigation Knock-out Knockout Mice Knowledge Lead Life Long-Term Effects Macaca fascicularis Macrocephaly Mediating Metabolic Metabolism Microscopic Mitochondria Modeling Morphology Motor Mus Muscle hypotonia Mutation N-acetylaspartate Neonatal Neuraxis Neurodegenerative Disorders Neurologic Neurons Oligodendroglia Organ Patients Peripheral Phenotype Physiological Prevention Psychomotor Performance Recombinant adeno-associated virus (rAAV)Research Role Safety Seizures Shapes Swelling Testing Therapeutic Tissues Toxic effect Transgenes Translations Urine adverse outcome base beta Actin cell type clinical application clinically relevant design disease phenotype gene replacement therapy gene therapy improved in vivo intravenous injection leukodystrophy loss of function mutation metabolome mouse model neuroimaging neuropathology nonhuman primate novel overexpression pre-clinical preclinical study promoter side effect treatment strategy vector white matter

项目摘要

ABSTRACT Canavan disease (CD) is a rare childhood leukodystrophy caused by autosomal recessive mutations in the aspartoacylase (ASPA) gene. Deficiency of ASPA in Canavan patients leads to the accumulation of N- Acetyl-Aspartic Acid (NAA), resulting in swelling and spongy degeneration of white matter in the brain. The clinical manifestations of this fatal disease include psychomotor retardation, muscular hypotonia, macrocephaly, head lag, seizures, and early death. Synthesis of NAA is carried out in the mitochondria of neurons by N-acetyltransferase-8-like (NAT8L) and hydrolyzed in oligodendrocytes by ASPA. Gene replacement therapy for ASPA deficiency is currently the most promising strategy for treating CD. Notably, we have recently achieved full therapeutic correction of the Canavan phenotype in the Aspa knockout (CD-KO) mouse model. A single intravenous injection of recombinant adeno-associated virus packaged with the human ASPA transgene (rAAV-hASPA) at early ages completely resolves neuropathology, resulting in treated animals that outperform wild-types in motor function tests. However, based on strong preliminary evidence, we now hypothesize that the CD phenotype presents a secondary etiology related to metabolism dysfunction. In addition, we recently revealed that overexpression of ASPA in wild type cells in vitro resulted in abnormal mitochondrial shape and function. These findings necessitate further preclinical investigations that focus on: 1) characterizing the possible toxicity of ASPA overexpression in cell types of the CNS and peripheral organs, 2) developing ASPA regulatory cassette(s) that can mimic endogenous physiological levels of ASPA to circumvent adverse effects that may exist due to treatment, and 3) determining the physiological and behavioral effects of ASPA overexpression using a clinically relevant non-human primate model. Our new research strategy now builds on our current promising progress and advances our goals for a safe and effective gene therapy for CD.

抽象的卡纳万病（CD）是一种罕见的儿童脑白质营养不良，由常染色体隐性突变引起天冬氨酸酰化酶 (ASPA) 基因。 Canavan 患者缺乏 ASPA 会导致 N- 积累乙酰天冬氨酸（NAA），导致大脑白质肿胀和海绵状变性。这这种致命疾病的临床表现包括精神运动迟缓、肌肉张力减退、巨大头畸形、头部迟滞、癫痫发作和过早死亡。 NAA的合成是在线粒体中进行的 N-乙酰转移酶-8 样 (NAT8L) 作用于神经元，并通过 ASPA 在少突胶质细胞中水解。基因 ASPA 缺乏症的替代疗法是目前治疗 CD 最有前途的策略。值得注意的是，我们最近在 Aspa 基因敲除 (CD-KO) 中实现了 Canavan 表型的完全治疗纠正鼠标模型。与人包装的重组腺相关病毒单次静脉注射 ASPA 转基因 (rAAV-hASPA) 在幼年时完全解决了神经病理学问题，从而获得了经过治疗的动物在运动功能测试中优于野生型。然而，基于强有力的初步证据，我们现在假设 CD 表型是与代谢功能障碍相关的次要病因。在此外，我们最近发现在体外野生型细胞中过度表达ASPA会导致异常线粒体的形状和功能。这些发现需要进一步的临床前研究，重点关注：1) 描述中枢神经系统和外周器官细胞类型中 ASPA 过度表达的可能毒性，2) 开发可以模拟 ASPA 内源性生理水平的 ASPA 调节盒规避治疗可能存在的不良影响，以及 3) 确定生理和使用临床相关的非人类灵长类动物模型研究 ASPA 过度表达的行为影响。我们的新研究战略现在建立在我们当前有希望的进展的基础上，并推进我们的目标，以实现安全和 CD 的有效基因治疗。