Development of a SYF2 antisense oligonucleotide treatment for ALS and FTD

开发治疗 ALS 和 FTD 的 SYF2 反义寡核苷酸

• 批准号: 10547625
• 负责人: Samuel V Alworth
• 金额: $ 149.25万
• 依托单位: ACURASTEM, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10547625
• 关键词: ALS patients; Address; Adult; Affect; Alleles; Amyotrophic Lateral Sclerosis; Antisense Oligonucleotides; Artificial Human Chromosomes; Automobile Driving; Autopsy; Bacterial Artificial Chromosomes; Bioinformatics; Biological Markers; Brain; C9ORF72; Cell Nucleus; Central Nervous System; Chemicals; Childhood; Complex; Cytoplasm; DNA Sequence Alteration; Data Set; Development; Disease; Dose; Evaluation; Exons; Frontotemporal Dementia; Generations; Genes; Genetic; Genetic Predisposition to Disease; Government; Human; Inclusion Bodies; Individual; Investigational Drugs; Investments; Maximum Tolerated Dose; Mus; Nerve Degeneration; Neuromuscular Junction; Neurons; Outcome; Pathologic; Pathology; Patients; Peripheral; Persons; Pharmaceutical Preparations; Phase; Phenotype; Population; Pre-Clinical Model; Probability; RNA; RNA Splicing; Rodent; Software Design; Spinal Cord; Spliceosomes; TDP-43 aggregation; Therapeutic; Therapeutic Index; Tissues; Toxic effect; Transgenic Mice; Work; amyotrophic lateral sclerosis therapy; causal variant; cohort; commercialization; design; frontotemporal lobar dementia amyotrophic lateral sclerosis; genetic approach; glial activation; immunogenicity; improved; in silico; lead candidate; loss of function; manufacturability; motor disorder; mouse model; new therapeutic target; novel therapeutic intervention; protein TDP-43; screening; sporadic amyotrophic lateral sclerosis; stathmin; success

Development of a SYF2 antisense oligonucleotide treatment for ALS and FTD Project Summary Amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) are complex diseases that result from many diverse genetic etiologies. Although therapeutic strategies that target specific causal mutations (e.g. C9ORF72 antisense oligonucleotides (ASOs)) may prove effective against individual forms of ALS or FTD, these approaches cannot address the vast majority of cases that have unknown genetic etiology. Moreover, given the large number of different genes that likely contribute to ALS and FTD and the fact that each genetic form is relatively rare, this strategy may be difficult to implement for all cases. Thus, there is a pressing need for new therapeutic strategies that rescue multiple forms of ALS and FTD, particularly those with unknown genetic etiologies. A hallmark pathological feature of ALS and FTD is the depletion of TAR DNA-binding protein 43 (TDP-43) from the nucleus of neurons in the brain and spinal cord to the cytoplasm where it aggregates into insoluble inclusion bodies in >95% of ALS cases and ~45% of FTD cases post mortem. While studies suggest that these neuronal TDP-43 aggregates drive neurodegeneration58, reduction in TDP-43 from the nucleus also alters the splicing or expression levels of more than 1,500 RNAs9, including disease hallmarks such as STMN2. Thus, both the loss of TDP-43 from the nucleus and its aggregation in the cytoplasm contribute to neurodegeneration, and it is critical to develop treatments that address both aspects of this challenging pathology. We found that suppressing the gene encoding the spliceosome-associated factor SYF2 alleviates TDP-43 aggregation and mislocalization, improves TDP-43 activity, and rescues C9ORF72 and sporadic ALS survival. Moreover, Syf2 suppression ameliorates neurodegeneration, neuromuscular junction loss, and motor dysfunction in TDP-43 mice. Mice with one loss-of-function copy of Syf2 are healthy and humans carrying a loss-of-function SYF2 allele are not affected by pediatric diseases or known to be affected by disease in adulthood. Thus, suppression of spliceosome-associated factors such as SYF2 is a promising and broadly-effective genetic target for ALS and FTD. Antisense oligonucleotides (ASOs) are an attractive approach for genetic targets in the central nervous system (CNS) like SYF2 because they can be injected directly into the spinal cord, achieve sustained target engagement throughout the CNS, and are less likely to cause peripheral toxicity. Leveraging our proprietary ASO design software, we assessed all possible SYF2 ASO sequences in silico and prioritized several hundred leads predicted to have enhanced stability and manufacturability, and reduced immunogenicity and off-target effects. The objective of this Direct to Phase 2 project is to identify a bona fide development candidate SYF2 ASO from among these leads for advancement in investigational new drug (IND)-enabling toxicity studies. The broad aims of the project are to establish the therapeutic index of Syf2 suppression in TDP-43 mice, assess SYF2 leads for potency, efficacy and tolerability in a large panel of ALS patient-derived neurons and a bacterial artificial chromosome human SYF2 transgenic mouse, and lastly to assess the leads in an exploratory toxicity study in rodents.

开发治疗 ALS 和 FTD 的 SYF2 反义寡核苷酸 项目概要 肌萎缩侧索硬化症 (ALS) 和额颞叶痴呆 (FTD) 是由多种原因引起的复杂疾病 不同的遗传病因。尽管针对特定因果突变的治疗策略（例如 C9ORF72 反义 寡核苷酸（ASO））可能被证明对个别形式的 ALS 或 FTD 有效，但这些方法无法解决 绝大多数病例的遗传病因不明。此外，考虑到大量不同的基因 可能会导致 ALS 和 FTD，而且每种遗传形式都相对罕见，这一策略可能很难实施 对所有情况实施。因此，迫切需要新的治疗策略来挽救多种形式的 ALS 和 FTD，特别是那些遗传病因不明的患者。 ALS 和 FTD 的一个标志性病理特征是 TAR DNA 结合蛋白 43 (TDP-43) 的缺失 脑和脊髓中的神经元细胞核进入细胞质，在细胞质中聚集成不溶性包涵体 尸检后，>95% 的 ALS 病例和 ~45% 的 FTD 病例。虽然研究表明这些神经元 TDP-43 聚集 驱动神经变性58，细胞核中 TDP-43 的减少也会改变超过 1,500 个 RNA9，包括 STMN2 等疾病标志。因此，TDP-43 从细胞核中丢失及其 细胞质中的聚集会导致神经退行性变，开发同时解决这两个问题的治疗方法至关重要 这种具有挑战性的病理学的各个方面。 我们发现抑制编码剪接体相关因子 SYF2 的基因可减轻 TDP-43 聚集 和错误定位，提高 TDP-43 活性，并挽救 C9ORF72 和偶发性 ALS 存活。此外，Syf2 抑制可改善 TDP-43 小鼠的神经变性、神经肌肉接头丧失和运动功能障碍。小鼠 携带 Syf2 功能缺失拷贝的人是健康的，携带功能缺失 SYF2 等位基因的人类不受 儿科疾病或已知在成年期受疾病影响的疾病。因此，剪接体相关因子的抑制 例如 SYF2 是 ALS 和 FTD 的一个有前景且广泛有效的基因靶点。 反义寡核苷酸 (ASO) 是中枢神经系统 (CNS) 遗传靶标的一种有吸引力的方法 与 SYF2 一样，因为它们可以直接注射到脊髓中，在整个过程中实现持续的目标参与 中枢神经系统，并且不太可能引起外周毒性。利用我们专有的 ASO 设计软件，我们评估了所有 计算机中可能的 SYF2 ASO 序列，并优先考虑了数百个预计具有增强稳定性的先导 可制造性，并降低免疫原性和脱靶效应。该 Direct to Phase 2 项目的目标是 从这些线索中识别出真正的开发候选者 SYF2 ASO，以促进研究性新药物的进展 药物 (IND) 毒性研究。该项目的主要目标是建立 Syf2 的治疗指数 TDP-43 小鼠中的抑制，评估 SYF2 在大量 ALS 中的效力、功效和耐受性 患者来源的神经元和细菌人工染色体人类 SYF2 转基因小鼠，最后评估引线 在啮齿动物的探索性毒性研究中。