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. .

抽象的 多聚谷氨酰胺 (polyQ) 疾病是最常见的遗传性神经退行性疾病之一 它们是由编码异常长的谷氨酰胺序列的扩展的 CAG 重复引起的。 在九种已知的 PolyQ 疾病中，脊髓小脑性共济失调 3 型 (SCA3)，也称为 马查多-约瑟夫病 (MJD) 是美国第二常见的疾病，也是世界上最常见的疾病。 SCA3 也是最常见的显性遗传性共济失调，其退化主要影响小脑， 脑干、黑质、丘脑和脊髓 在 SCA3 中，ATXN3 编码内的 CAG 重复。 通常包含 12 至 44 个 CAG 的序列已扩展到 60 至 87 个三联体，目前仅起到姑息作用。 可以使用直接针对扩展的 SCA3 的治疗策略。 mRNA，例如反义寡核苷酸（ASO），已经产生了有希望的结果，最近有两种药物被叫停。 基于 ASO 的治疗方法在针对多 Q 障碍（亨廷顿病）的临床试验中表明，需要 开发和评估多种治疗方式。 在此提案中，我们建议使用人工位点特异性 RNA 核酸内切酶 (ASRE) 技术 设计 CAG 重复特异性 RNA 核酸内切酶来破坏含有的扩展致病性 SCA3 RNA。 从 PUF 蛋白中分离出的 RNA 结合域，由一系列约 36 个氨基酸模块组成 在概念验证研究中，我们设计了 ASRE 帽子，可以识别一种特定的核糖核苷酸。 优先针对扩展的 ATXN3 RNA 在此 FastTrack 提案中，我们将寻求进一步增加。 在第一阶段，我们寻求评估针对 SCA3 的工程 ASRE 的可行性。 位于编码序列内的两个 SNP，经常将扩展的等位基因共分离以获取 基于 ASO 的治疗方法对这些 SNP 的观察显示出有希望的疗效结果。 这些研究将使 Enzerna 能够建立一系列基因疗法，从而提供精确的可能性 适合患者携带的特定 SCA3 疾病等位基因的药物方法。 将进行体外和体内研究，以评估 SCA3 相关表型异常的拯救 AAV 介导的候选 ASRE 疗法的递送。 从长远来看，与基因传递载体相结合，ASRE 提供了一种新的选择性策略 通过靶向 SCA3 (MJD) 的潜在基础，ASRE 可以降解致病性 ATXN3 转录本。 对这类无法治愈的人类疾病进行预防和/或治疗的可能性。 。