Development of allele-specific protein-based therapeutic targeting the pathogenic RNA associated with Spinocerebellar ataxia type 3

开发基于等位基因特异性蛋白的疗法，靶向与 3 型脊髓小脑共济失调相关的致病性 RNA

• 批准号: 10552829
• 负责人: JOSEPH C. RUIZ
• 金额: $ 25.91万
• 依托单位: ENZERNA BIOSCIENCES, LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-19 至 2024-08-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10552829
• 关键词: Affect; Alleles; Amino Acids; Animal Model; Antisense Oligonucleotides; Ataxia; Behavioral; Brain Stem; CAG repeat; Cell Line; Cells; Cerebellum; Clinical Trials; Code; Development; Disease; Dose; Endoribonucleases; Engineering; Fibroblasts; Gene Delivery; Gene Transfer; Glutamine; Human; Huntington Disease; In Vitro; Inherited; Link; Machado-Joseph Disease; Mediating; Messenger RNA; Modality; Molecular; Motor Activity; Neurodegenerative Disorders; Neuroglia; Neurons; Nucleotides; Pathogenicity; Patients; Phase; Phenotype; Preventive therapy; Protein Engineering; Proteins; Protocols documentation; RNA; RNA Binding; RNA Recognition Motif; RNA Sequences; Ribonucleotides; Series; Site; Spinal Cord; Stretching; Structure; Substantia nigra structure; Technology; Testing; Thalamic structure; Therapeutic; Transcript; Transgenic Animals; Transgenic Mice; Triplet Multiple Birth; Variant; base; cell type; curative treatments; delivery vehicle; design; efficacy study; genetic variant; human disease; human embryonic stem cell line; in vivo; induced pluripotent stem cell; knock-down; mouse model; nerve stem cell; novel strategies; novel therapeutics; palliative; polyglutamine; precision medicine; stem cells; symptom management; therapeutic gene; therapeutic target; transcriptome sequencing

ABSTRACT Polyglutamine (polyQ) diseases represent one of the more common classes of inherited neurodegenerative diseases. They are caused by expanded CAG repeats that encode abnormally long glutamine stretches in the disease proteins. Of the nine known polyQ disorders, Spinocerebellar Ataxia type 3 (SCA3), also known as Machado-Joseph disease (MJD), is the second most common in the US and the most common in the world. SCA3 is also the most common dominantly inherited ataxia with degeneration primarily affecting the cerebellum, brainstem, substantia nigra, thalamus and spinal cord. In SCA3, a CAG repeat within the ATXN3 coding sequences which normally harbors 12 to 44 CAGs is expanded to 60 to 87 triplets. Currently, only palliative therapeutics to manage symptoms are available. Therapeutic strategies directly targeting expanded SCA3 mRNAs, such as antisense oligonucleotides (ASO), have produced promising results. The recent halt of two ASO-based therapeutics in clinical trials for the polyQ disorder, Huntington’s Disease demonstrates, the need for the development and assessment of a diverse set of treatment modalities. In this proposal, we propose to use our Artificial SiteSpecific RNA Endonucleases (ASREs) technology to design CAG repeat specific RNA endonuclease to destroy expanded pathogenic SCA3 RNAs. ASREs contain RNA binding domains isolated from PUF proteins, which consist of a series of ~36 amino acid modules that recognize one specific ribonucleotide. In proof-of-concept studies, we have designed ASREs hat appear to preferentially target the expanded ATXN3 RNA. In this FastTrack proposal, we will seek to further increase therapeutic options for SCA3. In Phase I, we seek to assess the feasibility of engineering ASREs that can target two SNPs, located within the coding sequences, that frequently co-segregate the expanded allele to take advantage of the observations that ASO based therapeutics to these SNPs show promising efficacy results. These studies will enable Enzerna to build a portfolio of gene therapeutics that provide the possibility of precision medicine approaches appropriate for the specific SCA3 disease allele carried by the patient. In Phase, II, in vitro and in vivo studies will be conducted to assess rescue of SCA3-associated phenotypic anomalies after AAV-mediated delivery of the candidate ASRE therapeutics. In the long term, combined with gene delivery vectors, ASREs provide a new strategy for selective degradation of pathogenic ATXN3 transcripts. By targeting the underlying basis of SCA3 (MJD), ASREs provide the possibility of a preventative and/or curative therapy for this incurable class of human diseases. .

摘要 多聚谷氨酰胺（polyQ）疾病是遗传性神经退行性疾病中最常见的一类。 疾病它们是由扩增的CAG重复序列引起的，CAG重复序列编码异常长的谷氨酰胺片段， 疾病蛋白质在九种已知的polyQ疾病中，脊髓小脑共济失调3型（SCA 3），也称为 Machado-Joseph病（MJD）是美国第二常见的疾病，也是世界上最常见的疾病。 SCA 3也是最常见的显性遗传性共济失调，其变性主要影响小脑， 脑干、黑质、丘脑和脊髓。在SCA 3中，ATXN 3编码区内的CAG重复序列 通常含有12至44个CAG的序列被扩增至1060至87个三联体。目前，只有姑息 控制症状的治疗方法是可用的。直接靶向扩展SCA 3的治疗策略 mRNA，如反义寡核苷酸（阿索），已经产生了有希望的结果。最近，两个 基于ASO的治疗polyQ紊乱（亨廷顿病）的临床试验表明， 开发和评估一套多样化的治疗模式。 在这项提案中，我们建议使用我们的人工位点特异性RNA内切酶（ASRE）技术， 设计CAG重复序列特异性RNA内切酶以破坏扩增的致病性SCA 3 RNA。ASRE包含 从PUF蛋白分离的RNA结合结构域，其由一系列约36个氨基酸模块组成， 识别一种特定的核苷酸。在概念验证研究中，我们设计的ASRE似乎 优选靶向扩增的ATXN 3 RNA。在这个快速通道提案中，我们将寻求进一步增加 SCA 3的治疗选择。在第一阶段，我们寻求评估工程ASRE的可行性， 位于编码序列内的两个SNP，其经常共分离扩增的等位基因， 这是观察结果的优点，即针对这些SNP的基于阿索的治疗剂显示出有希望的功效结果。 这些研究将使Enzerna能够建立一个基因治疗组合，提供精确的可能性， 适合于患者携带的特定SCA 3疾病等位基因的药物方法。在第二阶段， 将进行体外和体内研究以评估SCA 3相关表型异常的挽救， 候选ASRE治疗剂的AAV介导的递送。 从长远来看，ASRE与基因递送载体相结合，为选择性表达提供了一种新的策略。 致病性ATXN 3转录物的降解。通过针对SCA 3（MJD）的底层基础，ASRE提供 对这类人类不治之症的预防性和/或治愈性治疗的可能性。 .