Silencing C9or72 with rAAV Mediated RNAi

用 rAAV 介导的 RNAi 沉默 C9or72

• 批准号: 9042441
• 负责人: Robert H. Brown
• 金额: $ 36.05万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9042441
• 关键词: Amyotrophic Lateral Sclerosis; Antibodies; Brain; C9ORF72; Cell Line; Cells; Cerebellum; Cerebrum; Clinical Trials; Development; Disease; Dose; Effectiveness; Exons; Exposure to; Fibroblasts; Frontotemporal Dementia; Future; Gene Transduction Agent; Genes; Genetic; Genetic Transcription; Goals; Health; Human; In Vitro; Intervention; Intravenous; Introns; Investigation; Lead; Mediating; Messenger RNA; Methods; MicroRNAs; Modeling; Motor Neurons; Mus; Mutation; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurons; Patients; Pattern; Peripheral; Phenotype; Primates; Proteins; RNA; RNA Interference; Recombinant adeno-associated virus (rAAV); Route; Safety; Sequence Homology; Series; Small RNA; Spinal Cord; Testing; Therapeutic; Toxic effect; Transcript; Transgenes; Transgenic Mice; Translating; Translations; Untranslated RNA; Viral Vector; Virus; Wild Type Mouse; autosomal dominant trait; clinical application; cytotoxic; effective therapy; gain of function; in vivo; knock-down; mRNA Precursor; member; mouse model; mutant; nervous system disorder; neuron loss; nonhuman primate; novel; research study; statistics; transcription factor

DESCRIPTION (provided by applicant): Amyotrophic lateral sclerosis (ALS) is a progressive, untreatable, uniformly fatal motor neuron disorder that sometimes develops concurrently with frontotemporal dementia (FTD). ALS is encountered in both sporadic (SALS) and familial (FALS) forms; about 10% of cases are transmitted as autosomal dominant traits. The cause of sporadic ALS is not known. Recently it was discovered that about 30-50% of FALS cases are caused by expansions of a non-coding hexanucleotide G4C2 expansion in the gene C9ORF72. These expansions are also detected in 10-20% of familial FTD, 10% of sporadic FTD and in ~5% of SALS. These statistics define the C9ORF72 G4C2 expansion as the most common cause of ALS. In the present study, we propose to use rAAV type Rh10 to introduce a microRNA to silence expression of the transcripts of C9ORF72 that harbor the offending G4C2 expansion. In Aim 1, we will screen, identify and optimize potential miRNAs in vitro. In Aim 2, we will further characterize the phenotype of our novel C9ORF72mutant transgenic mouse and investigate use of rAAVRh10-C9miRs to silencing the mutant C9 transgene and thereby mitigate the phenotype in this mouse. In Aim 3 we will investigate delivery and efficacy of the rAAVRh10-C9miRs in a non- human primate model as first step to translating this therapy to clinical application. Relevance: We believe that these studies will be highly relevant for several reasons. (1) There is a compelling unmet need for effective ALS treatments; this project focuses on the most common form of FALS, with applicability as well to some cases of SALS and FTD. (2) This investigation will develop the use of intrathecal rAAVRh10 as a gene therapy vector for the CNS; the intrathecal route is advantageous, permitting widespread delivery within the CNS with doses that are an order-of-magnitude lower than are required via intravenous delivery; and minimizing peripheral exposure to virus. (3) rAAV has not been used in human neurodegenerative disorders. The platform we propose to develop here should have broad applicability across a breadth of neurological disorders.

描述（由申请人提供）：肌萎缩性侧面硬化症（ALS）是一种进行性，不可治疗，均匀致命的运动神经元疾病，有时与额颞痴呆（FTD）同时发生。 ALS均以零星（萨尔）和家族性（fals）形式遇到；约有10％的病例以常染色体显性特征传播。零星ALS的原因尚不清楚。最近，发现大约30％至50％的fals病例是由基因C9ORF72中非编码六核苷酸G4C2扩展的扩展引起的。这些扩张在10-20％的家族性FTD，零星FTD的10％和约5％的sals中也被检测到。这些统计数据将C9ORF72 G4C2扩展定义为ALS的最常见原因。 在本研究中，我们建议使用RAAV型RH10引入microRNA，以使C9orf72的转录本具有沉默，该转录物具有有害的G4C2膨胀。在AIM 1中，我们将在体外筛选，识别和优化潜在的miRNA。在AIM 2中，我们将进一步表征我们的新型C9orf72Mutant转基因小鼠的表型，并研究使用Raavrh10-C9Mirs来沉默突变体C9转基因，从而减轻该小鼠中的表型。在AIM 3中，我们将研究Raavrh10-C9MIRS在非人类灵长类动物模型中的传递和功效，作为将这种疗法转化为临床应用的第一步。相关性：我们认为，由于多种原因，这些研究将高度相关。 （1）对有效的ALS治疗有令人信服的未满足的需求；该项目着重于最常见的伪造形式，以及某些萨尔和FTD的情况。 （2）这项研究将开发使用鞘内RAAVRH10作为中枢神经系统的基因治疗载体的使用；鞘内途径是有利的，允许在中枢神经系统内进行广泛的递送，其剂量比通过静脉输送低的量级降低了。并最大程度地减少外周暴露于病毒。 （3）RAAV尚未用于人类神经退行性疾病。我们建议在这里开发的平台应该在神经系统疾病的广度上具有广泛的适用性。