Ataxin-7 oligonucleotide knock-down to treat SCA7 retinal and cerebellar disease

Ataxin-7 寡核苷酸敲低治疗 SCA7 视网膜和小脑疾病

基本信息

批准号：
8774128
负责人：
ALBERT R LA SPADA
金额：
$ 49.51万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8774128
关键词：
Alleles Antisense Oligonucleotides Brain Stem CAG repeat Cataloging Catalogs Categories Cell Line Cerebellar Ataxia Cerebellar Diseases Cerebellar degeneration Cerebellum Clinical Trials Collaborations Contralateral Degenerative Disorder Disease Disease Progression Evaluation Eye Fibroblasts Functional disorder Funding Gene Expression Gene Expression Alteration Gene Expression Profile Gene Proteins Gene Silencing Genes Genetic Transcription Goals Histology Human In Vitro Inherited Injection of therapeutic agent Intervention Trial Knock-in Mouse Lead Methods Molecular Mus Mutation Nerve Degeneration Oligonucleotides Pathogenesis Pathology Patients Pharmacologic Substance Photoreceptors Pilot Projects Positioning Attribute Production Proteins RNA Reading Reagent Request for Proposals Retina Retinal Retinal Degeneration Retinal Diseases Retinal Photoreceptors Retinitis Pigmentosa Rhodopsin Role SCA7 protein Series Small Interfering RNA Techniques Testing Therapeutic Therapeutic Agents Toxic effect Translational Research Type 7 Spinocerebellar Ataxia Validation Vision Visual Wild Type Mouse Work base comparative efficacy in vivo knock-down member mouse model mutant polyglutamine preclinical study prevent programs protein aggregate protein aggregation protein expression protein misfolding public health relevance research study retinal neuron stem therapy development

项目摘要

DESCRIPTION (provided by applicant): Spinocerebellar ataxia type 7 (SCA7) is an autosomal dominant disorder that results in a cone-rod dystrophy form of retinal degeneration. The mutation inherited by SCA7 patients is a CAG/polyglutamine (polyQ) repeat expansion in the ataxin-7 gene. The SCA7 mutation results in production of a toxic polyQ-expanded ataxin-7 protein. Since the toxic gene product drives all subsequent disease pathology in SCA7, the most attractive therapeutic paradigm is to terminate expression of the mutant gene product. Here we propose continuation of a translational research program intended to yield therapeutic agents for SCA7 retinal disease, and potentially for related SCAs caused by CAG repeat expansion mutations. We are pursuing ataxin-7 "gene silencing" using a strategy that has already been successfully applied to reduce the expression of a toxic protein: oligonucleotide-based knock-down. To achieve the goals of this translational research program, we have created an academic-industrial partnership. In close collaboration with ISIS Pharmaceuticals, a company that specializes in anti-sense oligonucleotide (ASO) production and single-stranded siRNA (ss-siRNA) creation, we identified leads that markedly knock-down ataxin-7 expression. Based upon these in vitro results, we selected leads for in vivo validation, and performed pilot intra-vitreal injections in wild-type mice. These pilot intra-vitreal injections yielded robust (> 0%) knock-down of ataxin-7 expression for eyes injected with anti-ataxin-7 ASO, in comparison to eyes injected with diluent. Further pilot studies in SCA7 knock-in mice resulted in significant reductions in ataxin-7 protein aggregates in retinal photoreceptor cells in treated eyes. We also identified CAG-repeat targeting oligonucleotides for allele-selective targeting of polyQ-expanded ataxin-7 in fibroblast cell lines from patients. We therefore propose two aims: 1) Based upon efficacy of knock-down and ocular toxicity parameters, we will select one ataxin-7 ASO and perform intra-vitreal injections of ataxin-7 ASO in the right eye and control ASO/diluent in the let eye of SCA7 knock-in mice to determine if ASO delivery can prevent or ameliorate SCA7 retinal degeneration. Read-outs will include ataxin-7 aggregation, retinal histology, and visual function. Once we identify an efficacious ataxin-7 ASO, we will repeat the trial to confirm efficacy, compare gene expression alterations in ASO-treated and control-treated eyes, determine if ASO delivery in symptomatic SCA7 mice can stop or reverse disease progression, and evaluate the utility of the ataxin-7 ASO as a therapy for SCA7 cerebellar degeneration. 2) After we select a CAG-repeat targeting oligonucleotide, we will test if the oligonucleotide can prevent or ameliorate retinal degeneration in SCA7 knock-in mice, and then advance the oligonucleotide for confirmatory studies, transcriptome expression analysis, symptomatic intervention trials, and evaluation as a therapy for SCA7 neurodegeneration, applying similar experimental strategies as with the ataxin-7 ASO. At the conclusion of this project, we will be well positioned to proceed with IND-enabling studies with our industrial collaborator, as a prelude to a clinical trial in humn SCA7 patients.

描述（由申请人提供）：脊髓脑性共济失调7型（SCA7）是一种常染色体显性疾病，导致视网膜变性的锥杆性营养不良形式。 SCA7患者遗传的突变是CAG/聚谷氨酰胺（PolyQ）重复膨胀在ataxin-7基因中。 SCA7突变导致产生有毒PolyQ膨胀的ataxin-7蛋白。由于毒性基因产物在SCA7中驱动所有随后的疾病病理，因此最有吸引力的治疗范式是终止突变基因产物的表达。在这里，我们提出了一项转化研究计划的延续，该计划旨在产生SCA7视网膜疾病的治疗剂，并可能针对CAG重复扩张突变引起的相关SCA。我们正在使用已经成功应用以减少有毒蛋白的表达的策略来追求ataxin-7“基因沉默”：基于寡核苷酸的敲低。为了实现这项翻译研究计划的目标，我们建立了学术工业合作伙伴关系。在与ISIS Pharmaceuticals密切合作的情况下，该公司专门从事反义寡核苷酸（ASO）生产和单链siRNA（SS-SIRNA）的创作，我们确定了明显敲低ataxin-7表达的铅。基于这些体外结果，我们选择了用于体内验证的铅，并在野生型小鼠中进行了螺旋内注射。与注入稀释剂的眼睛相比，这些试点内注射剂对注射抗ATAXIN-7 ASO的眼睛的ataxin-7表达产生了稳健的（> 0％）的敲击。在SCA7敲门小鼠中进行进一步的初步研究，导致视网膜感光细胞中的Ataxin-7蛋白聚集体显着降低了处理的眼睛。我们还确定了靶向寡核苷酸的CAG重复靶向，用于在患者的成纤维细胞细胞系中PolyQ扩展的ataxin-7的等位基因选择性靶向。因此，我们提出了两个目的：1）基于敲低和眼毒性参数的功效，我们将选择一个ataxin-7 ASO，并在右眼和控制ASO/稀释剂中对SCA7敲击小鼠的右眼和控制ASO/稀释剂进行actaxin-7 ASO的注射，以防止Aso Aso Expraiment saveral sca Aso sca sca7 sca7 sca7 sca7 sca7 sca7 sca7 sca7 sca7。读出将包括Ataxin-7聚集，视网膜组织学和视觉功能。一旦我们确定了有效的ataxin-7 ASO，我们将重复该试验以确认功效，比较ASO治疗和对照治疗的眼睛中的基因表达改变，确定在有症状的SCA7小鼠中的ASO递送是否可以停止或反向疾病进展，并评估Ataxin-7 ASO ASO ASO ASE治疗的效用。 2）在选择靶向寡核苷酸的CAG重复点之后，我们将测试寡核苷酸是否可以预防或改善SCA7敲入小鼠的视网膜退化，然后在验证性研究中提高寡核苷酸，以进行验证性研究，转录组表达分析，症状分析，症状介入试验以及对SCA7 NEURODEN的策略以及sca7 neurody-a sca7-Neurody-a sca7 neurodenies-a sca7-a sca7-a sca7-a sca 7 Aso。在该项目结束时，我们将有能力与我们的工业合作者进行辅助研究，作为Humn SCA7患者临床试验的前奏。