In vivo modelling and therapy development for stathmin-2 loss in TDP-43 proteinopathies

TDP-43 蛋白病中 stathmin-2 缺失的体内建模和治疗开发

• 批准号: 10317404
• 负责人: Don W Cleveland
• 金额: $ 250.73万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10317404
• 关键词: Adult; Affect; Alleles; Alzheimer' s disease patient; Amyotrophic Lateral Sclerosis; Animal Model; Animals; Antisense Oligonucleotides; Axon; Axotomy; Binding; Binding Proteins; Binding Sites; C9ORF72; Chronic; Cognition Disorders; Collaborations; Cytoplasm; Disease; Frontotemporal Dementia; Genes; Genetically Engineered Mouse; Human; Industrialization; Inherited; Injections; Introns; Lead; Maintenance; Mediating; Messenger RNA; Modeling; Molecular; Motor; Motor Neurons; Mus; Mutation; Natural regeneration; Nerve Degeneration; Nerve Regeneration; Nervous system structure; Neurodegenerative Disorders; Neuromuscular Junction; Neurons; Nuclear; PGRN gene; Pathogenesis; Pathologic; Pathology; Patients; Phenotype; Polyadenylation; Pre-Clinical Model; Proteins; RNA; RNA Processing; RNA Splicing; RNA-Binding Proteins; Regenerative capacity; Rodent; Role; Site; Spinal Cord; Testing; Therapeutic; Tissues; Transcript; alpha Tubulin; axon regeneration; base; falls; familial amyotrophic lateral sclerosis; frontotemporal lobar dementia-amyotrophic lateral sclerosis; genome editing; granulin; humanized mouse; in vivo; in vivo Model; in vivo evaluation; induced pluripotent stem cell; insight; loss of function; mRNA Precursor; model development; motor deficit; mouse genome; mouse model; nervous system disorder; neuronal growth; novel therapeutic intervention; premature; prevent; programs; protein TDP-43; restoration; small hairpin RNA; sporadic amyotrophic lateral sclerosis; stathmin; therapeutic development; therapeutic effectiveness; therapeutic evaluation; therapy development

PROJECT SUMMARY TDP-43 proteinopathies represent a set of neurological disorders characterized by relocalization of the RNA- binding protein TDP-43 from its native nuclear compartment to the cytoplasm where it accumulates into aggregates. TDP-43 pathology is found in more than 95% of patients with amyotrophic lateral sclerosis (ALS), approximately half of frontotemporal dementia (FTD) cases, and more than 30% of Alzheimer’s disease patients. TDP-43 is involved in fundamental RNA processing activities and binds thousands of transcripts to regulate their expression, splicing and transport. We and others have recently identified a critical role for TDP-43 in regulating the expression of the neuronal growth-associated factor stathmin-2, a tubulin-binding protein involved in axon outgrowth and regeneration. TDP-43 disruption induces truncation (by aberrant splicing and premature polyadenylation) of stathmin-2 pre-mRNA, thereby silencing stathmin-2 when TDP-43 nuclear levels fall. Stathmin-2 is suppressed in affected neurons from the vast majority of ALS/FTD patients, and the neuronal regeneration capacity of iPSC-derived motor neurons with TDP-43 depletion can be rescued by increasing levels of STMN2. Notably, while stathmin-2 is the human mRNA most affected by reduction in TDP-43, the stahmin-2 pre-mRNA is neither bound nor regulated by TDP-43 in rodents, a major caveat for animal modeling of TDP-43 proteinopathies. Here we propose to use newly generated mouse models to determine whether inactivated (Aim 1) or humanized stathmin-2 (Aim 2) alleles synergize with TDP-43, progranulin, or C9ORF72 ALS/FTD mutations to drive motor neuron or cognitive disease. While evidence supports that stathmin-2 is essential for axonal regeneration, the contribution of stathmin-2 loss in neurodegeneration remains to be determined. We propose an ambitious program to establish the impact of in vivo stathmin-2 reduction in TDP-43 proteinopathies. Mouse models generated in this project will recapitulate a major molecular alteration associated with TDP-43 proteinopathy and represent an important platform for therapeutic development in ALS/FTD. In addition, recognizing that restoration of stathmin-2 level is an attractive strategy with broad implications in neurodegenerative diseases, we will test the therapeutic potential of antisense oligonucleotides (ASO) that prevent aberrant splicing of Stmn2 (Aim 3). This collaborative effort has the potential to uncover new insights on the contribution of stathmin-2 loss in neurodegeneration, to generate new preclinical models, and to develop a novel therapeutic strategy for ALS and FTD.

项目概要 TDP-43 蛋白病代表一组以 RNA 重新定位为特征的神经系统疾病。 结合蛋白 TDP-43 从其天然核区室到细胞质，并在细胞质中积累 TDP-43 病理学存在于超过 95% 的肌萎缩侧索硬化症 (ALS) 患者中， 大约一半的额颞叶痴呆 (FTD) 病例和超过 30% 的阿尔茨海默病患者。 TDP-43 参与基本的 RNA 加工活动并结合数千个转录物来调节它们的 我们和其他人最近发现了 TDP-43 在调节中的关键作用。 神经元生长相关因子 stathmin-2 的表达，一种参与轴突的微管蛋白结合蛋白 TDP-43 的生长和再生破坏会导致截短（通过异常剪接和过早发生）。 Stathmin-2 前 mRNA 的聚腺苷酸化），从而在 TDP-43 核水平下降时沉默 Stathmin-2。 Stathmin-2 在绝大多数 ALS/FTD 患者受影响的神经元中受到抑制，并且神经元 提高 TDP-43 水平可以挽救 iPSC 来源的运动神经元的再生能力 值得注意的是，虽然 stathmin-2 是受 TDP-43 减少影响最大的人类 mRNA，但 stathmin-2 的影响最大。 在啮齿类动物中，前 mRNA 既不结合也不受 TDP-43 调节，这是 TDP-43 动物模型的一个主要警告 在这里，我们建议使用新生成的小鼠模型来确定是否失活（Aim） 1) 或人源化 stathmin-2 (目标 2) 等位基因与 TDP-43、颗粒体蛋白前体或 C9ORF72 ALS/FTD 协同作用 而有证据表明，stathmin-2 对于驱动运动神经元或认知疾病至关重要。 轴突再生中，stathmin-2 缺失在神经退行性疾病中的作用仍有待确定。 提出了一项雄心勃勃的计划，以确定体内 stathmin-2 减少对 TDP-43 蛋白病的影响。 该项目生成的小鼠模型将重现与 TDP-43 相关的主要分子改变 蛋白质病是 ALS/FTD 治疗开发的重要平台。 认识到恢复 Stathmin-2 水平是一项有吸引力的策略，具有广泛的影响 神经退行性疾病，我们将测试反义寡核苷酸（ASO）的治疗潜力 防止 Stmn2 的异常剪接（目标 3）。 Stathmin-2 缺失在神经退行性疾病中的贡献，生成新的临床前模型，并开发 ALS 和 FTD 的新治疗策略。