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ALS和FTD的SYF2反义寡核苷酸处理 项目摘要 肌萎缩性外侧硬化症（ALS）和额颞痴呆（FTD）是由许多人引起的复杂疾病 多样化的遗传病因。尽管针对特定因果突变的治疗策略（例如C9orf72反义 寡核苷酸（ASOS）可能证明对ALS或FTD的个体形式有效，这些方法无法解决 绝大多数具有未知遗传病因的病例。而且，鉴于大量不同的基因 可能有助于ALS和FTD，并且每个遗传形式相对较少，这一事实可能很难 所有情况下实施。因此，迫切需要采取新的治疗策略来挽救多种ALS的形式 和FTD，特别是那些遗传病因未知的人。 ALS和FTD的标志性病理特征是tar DNA结合蛋白43（TDP-43）的耗竭 大脑中神经元的核和脊髓到细胞质的脊髓核，在那里它聚集成不溶性包容物体 > 95％的ALS病例和约有45％的FTD病例。研究表明这些神经元TDP-43骨料 驱动神经变性58，核中TDP-43的降低也改变了超过超过的剪接或表达水平 1,500 RNAS9，包括诸如STMN2之类的疾病标志。因此，核及其损失TDP-43及其损失 细胞质中的聚集有助于神经退行性，开发既定的治疗方法至关重要 这种具有挑战性的病理的方面。 我们发现抑制编码剪接体相关因子SYF2的基因减轻了TDP-43聚集 和错误的定位，改善TDP-43活性，并挽救C9orf72和零星的ALS生存。此外，SYF2 抑制可改善TDP-43小鼠的神经变性，神经肌肉连接损失和运动功能障碍。老鼠 SYF2的功能丧失副本是健康的，携带功能丧失SYF2等位基因的人不会受到影响 小儿疾病或已知成年后受疾病影响。因此，抑制剪接体相关因子 例如SYF2是ALS和FTD的有前途且广泛的遗传靶标。 反义寡核苷酸（ASOS）是中枢神经系统（CNS）的遗传靶标的有吸引力的方法 像SYF2一样，因为它们可以直接注入脊髓，在整个过程中实现持续的目标参与 中枢神经系统，不太可能引起外围毒性。利用我们的专有ASO设计软件，我们评估了所有 在计算机中可能的SYF2 ASO序列，并优先考虑数百个铅的稳定性和 可制造性，降低免疫原性和脱靶效应。直接到第2阶段项目的目的是 从这些铅中识别出真正的开发候选候选人SYF2 ASO 药物（IND） - 增强毒性研究。该项目的广泛目的是建立SYF2的治疗指数 在TDP-43小鼠中抑制，评估SYF2导致大型ALS面板中的效力，功效和耐受性 患者衍生的神经元和细菌人造染色体人类SYF2转基因小鼠，最后评估铅 在啮齿动物的探索性毒性研究中。