Targeting STAU1 for TDP-43 proteinopathies

靶向 STAU1 治疗 TDP-43 蛋白病

• 批准号: 10512615
• 负责人: Stefan M. PULST
• 金额: $ 38.37万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10512615
• 关键词: ALS patients; Age; Aging; Amyotrophic Lateral Sclerosis; Antisense Oligonucleotides; Autophagocytosis; Bacterial Artificial Chromosomes; Biological Assay; Cells; Chemicals; Chemistry; Data; Development; Disease; Disease model; Dose; Fibroblasts; Financial Hardship; Frequencies; Future; Genes; Genetic; Grant; Health; High Pressure Liquid Chromatography; Histologic; Human; In Vitro; Injections; Investigation; LY6E gene; Lead; Measures; Methods; Modeling; Modification; Molecular; Motor; Motor Cortex; Motor Neurons; Mus; Nerve Degeneration; Neurodegenerative Disorders; Pathologic; Patients; Phase; Phenotype; Population; PrP; Preparation; Proteins; Purkinje Cells; Solubility; Spinal Cord; Structure-Activity Relationship; Synapses; Therapeutic; Toxic effect; Transgenic Mice; United States National Institutes of Health; Validation; Vertebral column; Work; acute toxicity; base; efficacy study; efficacy testing; experience; improved; in vivo; in vivo Model; knock-down; molecular phenotype; motor behavior; mouse model; nervous system disorder; neurodegenerative phenotype; phase I trial; phosphorothioate; protein TDP-43; protein aggregation; protein biomarkers; screening; sporadic amyotrophic lateral sclerosis; success; therapeutic target; uptake

Project Summary/Abstract Our U01 CREATE grant supported development of BIIB105, an ATXN2 antisense oligonucleotide (ASO) for treatment that is now in a phase 1 trial for amyotrophic lateral sclerosis (ALS). We are now proposing to expand the ATXN2 target space by developing ASOs to STAU1. STAU1 interacts with ATXN2, and is overabundant in multiple in vitro and in vivo models of neurodegeneration. Our proof-of-concept data showed positive PD efficacy, in that lowering STAU1 modifies disease-related phenotypes. ATXN2[Q127] SCA2 mice haploinsufficient for Stau1 had normalized cerebellar gene and autophagy protein abundances, ATXN2 protein aggregates were eliminated, and motor behavior was improved. In addition, the abnormally slow intrinsic Purkinje cell (PC) firing frequency in SCA2 mice is restored following Stau1 ASO treatment. STAU1 expands the ATXN2 target space and may prove more effective as a therapeutic target for ALS and other TDP-43 proteinopathies as its protein abundance is increased 3-4 fold in multiple in vitro and in vivo neurodegenerative models including sporadic ALS spinal cord. We have already screened for ASOs targeting STAU1, identifying 10 top leads that lower STAU1 expression by 90-99% in SCA2 patient fibroblasts, including two potent ASOs that also target mouse Stau1 invaluable for in vivo efficacy studies in disease models. Our overall objective is to optimize ASO sequence and chemistry for lowering STAU1 expression, and to refine an in vivo efficacy models for characterizing target engagement and PD/PK parameters in vivo. Currently, no therapeutics exist that significantly modify sporadic ALS. STAU1 is overabundant in multiple neurodegenerative disease models, and Stau1 knockdown by genetic interaction improves in vivo phenotypes. The R61 phase of our study will span one year and is organized in two aims: Aim 1 will prepare sequence optimized chemically-modified ASOs for efficacy testing in BAC-STAU1 mice and Prp-TDP43Q331K mice. Aim 2 refines the use of Prp-TDP43Q331K mice for characterizing efficacy of STAU1 ASO, resulting in optimal doses required for effective STAU1 knockdown and phenotype modification. The R33 phase spans the remaining two years of the study and is comprised of two additional aims: Aim 3 will demonstrate that lead ASOs modify TDP-43 abundance & solubility, normalizes autophagy markers, histological phenotypes and motor neuron counts and motor behavior of Prp-TDP43Q331K mice. Aim 4 will determine ASO effects for improving survival for three most efficacious lead ASOs using Prp- TDP43WTxQ331K double transgenic mice. The terminal milestone for this study will be validation of in vivo efficacy for three optimized ASOs with long duration of action for Staufen knockdown, and efficacy data for additional ASO sequences useful or future investigation. The crucial objective is the identification of lead STAU1 ASOs that modify an ALS neurodegenerative phenotype suitable for advancing to a URGenT or Blueprint Neurotherapeutics Network (BPN) application to the NIH. Ultimately, our study will significantly expand therapeutic targets for ALS and other neurodegenerative diseases.

项目总结/摘要 我们的U01 CREATE资助支持BIIB105的开发，BIIB105是一种ATXN2反义寡核苷酸（ASO），用于 该疗法目前正处于肌萎缩侧索硬化症（ALS）的1期临床试验中。我们现在建议扩大 ATXN2通过向STAU1发展ASO来瞄准空间。STAU1与ATXN2相互作用，并且在 多种体外和体内神经变性模型。我们的概念验证数据显示， 有效性，因为降低STAU1修饰疾病相关表型。ATXN2 [Q127] SCA2小鼠 Stau 1单倍型不足的患者小脑基因和自噬蛋白丰度正常化，ATXN2蛋白 聚集体被消除，运动行为得到改善。此外，异常缓慢的内在 SCA2小鼠中的浦肯野细胞（PC）放电频率在Stau 1 ASO处理后恢复。STAU1扩展了 ATXN2靶向空间，并且可能证明作为ALS和其他TDP-43的治疗靶点更有效 在多种体外和体内神经退行性疾病中， 包括散发性ALS脊髓模型。我们已经筛选了针对STAU 1的ASO， 在SCA2患者成纤维细胞中将STAU1表达降低90 - 99%的10种主要先导化合物，包括两种有效的ASO 其也靶向小鼠Stau 1，对于疾病模型中的体内功效研究非常宝贵。我们的总体目标是 优化用于降低STAU1表达的ASO序列和化学，并完善体内功效模型 用于表征体内靶标接合和PD/PK参数。目前，不存在治疗方法， 显著改善散发性ALS。STAU1在多种神经退行性疾病模型中过量， 通过遗传相互作用敲低Stau1可改善体内表型。我们研究的R61阶段将跨越一个 目标1将制备序列优化的化学修饰的ASO， 在BAC-STAU1小鼠和Prp-TDP43Q331K小鼠中的功效测试。目的2：改良Prp-TDP43Q331K小鼠的使用 用于表征STAU1 ASO的功效，得到有效STAU1敲低所需的最佳剂量 和表型修饰。R33阶段涵盖研究的剩余两年，包括 两个额外的目标：目标3将证明，铅ASO修改TDP-43丰度和溶解度， Prp-TDP43Q331K的自噬标志物、组织学表型和运动神经元计数以及运动行为 小鼠目标4将确定使用Prp的三种最有效的电极导线ASO在改善生存期方面的ASO效应， TDP43WTxQ331K双转基因小鼠。本研究的最终里程碑将是体内疗效验证 对于Staufen敲低具有长作用持续时间的三种优化的ASO，以及对于额外的 ASO序列有用或未来调查。关键目标是确定STAU 1的主要ASO 其修饰ALS神经变性表型，适合于进展到URGenT或蓝图 神经治疗学网络（BPN）在NIH的应用。最终，我们的研究将大大扩展 ALS和其他神经退行性疾病的治疗靶点。