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

基本信息

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
项目类别：
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&apos 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

项目摘要

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蛋白质病代表了一组由RNA-重新定位的神经系统疾病结合蛋白TDP-43从其天然核室到细胞质的结合，并在其中积聚聚合。在超过95％的肌萎缩性侧索硬化症患者（ALS）中发现了TDP-43病理学大约一半的额颞痴呆（FTD）病例，而阿尔茨海默氏病患者的30％以上。 TDP-43参与了基本的RNA处理活动，并结合了数千个成绩单以调节其表达，剪接和运输。我们和其他人最近确定了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，一种与轴突有关的微管蛋白结合蛋白生长和再生。 TDP-43破坏引起截断（通过异常剪接和过早 Stathmin-2前MRNA的聚腺苷酸化），从而使TDP-43核水平下降时沉默的Stathmin-2。 STATHMIN-2在绝大多数ALS/FTD患者的受影响神经元中受到抑制，神经元 IPSC衍生的运动神经元具有TDP-43耗竭的再生能力可以通过增加水平来检索 STMN2。值得注意的是，尽管Stathmin-2是受TDP-43减少影响的人类mRNA，但Stahmin-2 前MRNA既不受TDP-43的结合，也不受TDP-43的调节，这是TDP-43动物建模的主要警告蛋白质病。在这里，我们建议使用新生成的鼠标模型来确定是否灭活（AIM 1）或人源化的Stathmin-2（AIM 2）等位基因与TDP-43，ProgramRunulin或C9orf72 ALS/FTD协同作用驱动运动神经元或认知疾病的突变。虽然证据支持Stathmin-2对于轴突再生，神经退行性中的Stathmin-2损失的贡献尚待确定。我们提出了一个雄心勃勃的计划，以建立在TDP-43蛋白质中减少体内Stathmin-2的影响。该项目中产生的小鼠模型将概括与TDP-43相关的主要分子改变蛋白质病代表了ALS/FTD中治疗性发育的重要平台。此外，认识到恢复Stathmin-2水平是一种有吸引力的策略，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（AIM 3）。这种合作的努力有可能发现有关的新见解 Stathmin-2在神经变性中的贡献，生成新的临床前模型，并开发一个 ALS和FTD的新型治疗策略。