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.

项目摘要 TDP-43蛋白病代表一组以RNA-43的重新定位为特征的神经系统疾病。 结合蛋白TDP-43从其天然的核区室到细胞质，在细胞质中积累成 集料. TDP-43病理学在超过95%的肌萎缩侧索硬化症（ALS）患者中发现， 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，并结合数千种转录物以调节其表达。 表达、剪接和运输。我们和其他人最近发现了SDP-43在调节方面的关键作用 神经生长相关因子stathmin-2的表达，stathmin-2是一种参与轴突生长的微管蛋白结合蛋白， 生长和再生。TDP-43破坏诱导截短（通过异常剪接和过早的 多聚腺苷酸化），从而当TDP-43核水平下降时沉默stathmin-2。 Stathmin-2在来自绝大多数ALS/FTD患者的受影响神经元中受到抑制，并且神经元 具有TDP-43耗尽的iPSC衍生的运动神经元的再生能力可以通过增加水平来挽救 关于STMN 2值得注意的是，虽然stahmin-2是受TDP-43减少影响最大的人mRNA，但stahmin-2是受TDP-43减少影响最大的人mRNA。 在啮齿类动物中，前mRNA既不受TDP-43的结合也不受其调节，这是TDP-43动物模型的一个主要警告 蛋白质病在这里，我们建议使用新生成的小鼠模型，以确定是否灭活（目的 1)或人源化stathmin-2（Aim 2）等位基因与TDP-43、颗粒蛋白前体或C9 ORF 72 ALS/FTD协同作用 导致运动神经元或认知疾病的突变。虽然有证据表明stathmin-2对于 轴突再生，stathmin-2丢失在神经变性中的作用仍有待确定。我们 提出了一个雄心勃勃的计划，以建立体内stathmin-2减少在TDP-43蛋白病中的影响。 本项目中产生的小鼠模型将重现与TDP-43相关的主要分子改变 蛋白质病，代表了ALS/FTD治疗开发的重要平台。此外，本发明还提供了一种方法， 认识到恢复stathmin-2水平是一项有吸引力的战略， 神经退行性疾病，我们将测试反义寡核苷酸（阿索）的治疗潜力， 防止Stmn 2的异常剪接（目的3）。这种合作努力有可能揭示新的见解， stathmin-2缺失在神经退行性变中的作用，以产生新的临床前模型，并开发一种新的 ALS和FTD的新治疗策略。