Therapeutic strategies for microsatellite expansion diseases using RNA targeting

利用 RNA 靶向治疗微卫星扩增疾病的策略

基本信息

批准号：
10588064
负责人：
MAURICE SCOTT SWANSON
金额：
$ 65.74万
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：
2017
资助国家：
美国
起止时间：
2017-09-22 至 2027-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10588064
关键词：
3&apos Untranslated Regions Acceleration Address Adult Alleles Antisense Oligonucleotides Attenuated BCAR1 gene Binding Brain C9ORF72 CUG repeat Cell model Cells Data Dependovirus Development Disease Disease model Engineering Evaluation Event Exons Gene Expression Genetic Transcription Genome Health Human Human Genome Huntington Disease Immunity Individual Knock-in Knock-in Mouse Link Mediating Microsatellite Repeats Modality Modeling Molecular Mus Muscle Muscle Development Muscular Dystrophies Mutation Myoblasts Myocardium Myotonic dystrophy type 1 Neurocognitive Deficit Neurodegenerative Disorders Neuromuscular Diseases Organoids Pathology Poly A Polyadenylation Predisposition Proteins RNA RNA Processing RNA Splicing RNA delivery RNA-targeting therapy Recombinants Reporter Risk Safety Short Tandem Repeat Signal Transduction Skeletal Muscle Small Interfering RNA Small Nuclear RNA Specificity Spinocerebellar Ataxias Spliceosomes System Tandem Repeat Sequences Testing Therapeutic Therapeutic Agents Time Tissues Toxic effect Transcript U1 small nuclear RNA Work adaptive immune response adeno-associated viral vector brain abnormalities design disorder control effective therapy flexibility frontotemporal lobar dementia amyotrophic lateral sclerosis gain of function gene therapy human stem cells humanized mouse in utero in vivo innovation insight knock-down lead candidate mouse model mutant nervous system disorder novel novel therapeutic intervention safety testing small hairpin RNA societal costs therapeutic RNA therapeutic candidate therapy development

项目摘要

PROJECT SUMMARY Microsatellites, also known as simple sequence repeats or short tandem repeats (STRs), are 2-10 bp tandem sequence repeats that occur throughout the human genome. Germ-line expansions of these repeats beyond a critical threshold are associated with more than 50 neurological, neurodegenerative and neuromuscular disorders, including Huntington disease, C9orf72-linked amyotrophic lateral sclerosis/ frontotemporal dementia, several types of spinocerebellar ataxias, as well as myotonic dystrophy types 1 and 2 (DM1/2). Although the complete range of molecular features associated with these diseases varies among these conditions (and often are unknown), cellular and mouse models demonstrate that RNA-mediated toxicity is a major factor. Thus, suppression of mutant RNA levels is expected to address all associated pathologies. While numerous strategies exist to attenuate gene expression via targeted destruction of RNA, all have limitations relating to their mode of administration and tissue targeting (e.g. antisense oligonucleotides, siRNAs), or carry the risk of toxicity caused by adaptive immune responses or pre-existing immunity to the therapeutic agent (systems based delivery of RNA-targeting Cas proteins). Here we develop novel non-immunogenic RNA targeting system compatible with delivery by recombinant adeno-associated viral vectors (rAAVs) that support safe and long-lasting expression. Our platform is based on human spliceosomal RNAs, which in preliminary data we show can be engineered to target and degrade STR-containing transcripts. As a proof-of principle for the therapeutic potential of this system, we focus on DM1, the most common form of adult-onset most common adult-onset muscular dystrophy. We have developed a novel human stem cell based organoid model that, for the first time, provides insight into the molecular basis of the severe neurocognitive deficits associated with DM1. We will use this model to test the safety and efficacy of our RNA-targeting systems in conjunction with a novel mouse model that we generate and that we hope will recapitulate the multi-tissue pathology of DM1. If successful, our work will provide a flexible therapeutic RNA-targeting based platform for treatment of STR-associated diseases.

项目概要微卫星，也称为简单序列重复或短串联重复 (STR)，是 2-10 bp 串联的整个人类基因组中出现的序列重复。这些重复的种系扩展超出了临界阈值与 50 多种神经、神经退行性和神经肌肉相关疾病，包括亨廷顿病、C9orf72 相关的肌萎缩侧索硬化症/额颞叶痴呆、多种类型的脊髓小脑共济失调，以及 1 型和 2 型强直性肌营养不良 (DM1/2)。虽然与这些疾病相关的完整分子特征在这些条件之间有所不同（并且通常未知），细胞和小鼠模型表明 RNA 介导的毒性是一个主要因素。因此，抑制突变 RNA 水平有望解决所有相关的病理问题。虽然策略众多存在的目的是通过靶向破坏 RNA 来减弱基因表达，但它们都存在与其作用模式相关的局限性。给药和组织靶向（例如反义寡核苷酸、siRNA），或存在引起毒性的风险通过适应性免疫反应或对治疗剂的预先存在的免疫（基于系统的递送） RNA 靶向 Cas 蛋白）。在这里，我们开发了新型非免疫原性 RNA 靶向系统，兼容通过支持安全和持久表达的重组腺相关病毒载体（rAAV）进行递送。我们的平台基于人类剪接体 RNA，我们在初步数据中显示，它可以被设计为靶向并降解含有 STR 的转录本。作为该系统治疗潜力的原理证明，我们重点关注 DM1，这是成人发病最常见的成人发病性肌营养不良症的最常见形式。我们开发了一种新型的基于人类干细胞的类器官模型，该模型首次提供了对与 DM1 相关的严重神经认知缺陷的分子基础。我们将使用这个模型来测试我们的 RNA 靶向系统与我们生成的新型小鼠模型相结合的安全性和有效性我们希望能够重现 DM1 的多组织病理学。如果成功的话，我们的工作将提供灵活的用于治疗 STR 相关疾病的基于 RNA 靶向的治疗平台。