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Determining stathmin-2 function and potential as a therapeutic target in ALS/FTD

确定 Stathmin-2 的功能和作为 ALS/FTD 治疗靶点的潜力

基本信息

批准号：
10370327
负责人：
Don W Cleveland
金额：
$ 79.73万
依托单位：
LUDWIG INSTITUTE FOR CANCER RES LTD
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10370327
关键词：
Affect Amyotrophic Lateral Sclerosis Antisense Oligonucleotides Axon Axotomy Binding Binding Sites C9ORF72 CRISPR screen Cells Cognition Disorders Custom Cytoplasmic Protein DNA-Binding Proteins Dementia Denervation Disease Flow Cytometry Frontotemporal Dementia Genes Genetic Genetic Screening Growth Cones Guide RNA Hereditary Disease High-Throughput RNA Sequencing Human Induced pluripotent stem cell derived neurons Inherited Injections Introns Libraries Link Maintenance Messenger RNA Microtubule Stabilization Microtubules Motor Neuron Disease Motor Neurons Mus Muscle Mutation Natural regeneration Neurobiology Neuromuscular Junction Neurons Nuclear Optical Methods Optics Outcome Paralysed Pathogenesis Pathology Polyadenylation Post-Translational Protein Processing Proteins RNA RNA Sequences RNA Splicing Role Site Synapses Therapeutic Tubulin Variant axon growth axon regeneration base beta Tubulin dimer frontotemporal lobar dementia-amyotrophic lateral sclerosis genome editing genome-wide induced pluripotent stem cell insight loss of function mRNA Precursor motor neuron function nerve supply neuron regeneration neuronal cell body palmitoylation premature prevent protein TDP-43 repaired restoration small hairpin RNA sporadic amyotrophic lateral sclerosis stathmin therapeutic target

项目摘要

Cytoplasmic protein accumulations of the RNA/DNA binding protein TDP-43 are found in affected neurons in almost all instances of amyotrophic lateral sclerosis (ALS) and approximately 50% of frontotemporal dementia (FTD). Nuclear clearance of TDP-43 has been widely observed in affected neurons in sporadic ALS/FTD, evidence strongly supporting the proposal that TDP-43 loss of function is a key aspect of disease mechanism underlying ALS/FTD pathogenesis. We have identified that the mRNA encoding stathmin-2 is 1) an essential factor for axonal regeneration of axotomized iPSC-derived motor neurons and 2) the mRNA most affected by reduction in TDP-43 function, with a striking loss from motor neurons in sporadic ALS and inherited disease from GGGGCC expansion in C9orf72. Stathmin-2 is an abundant, direct binding partner of α/β-tubulin dimers in neuronal perikarya, axons, growth cones, and synapses, including neuromuscular junctions (NMJs). Using genome editing, we will identify the mechanism of TDP-43-dependent premature polyadenylation/cryptic splicing that suppresses stathmin-2 synthesis when TDP-43 levels are lowered. We will use genome editing of induced pluripotent stem cells (iPSCs) derived human motor neurons grown in compartmented chambers (that separate neuronal cell bodies, axons and growth cones) to determine how stathmin-2 functions in a) motor neuron maintenance/repair, b) axonal microtubule stabilization and/or dynamics, c) neuromuscular junction formation and/or stabilization, and d) how palmitoylation of stathmin-2 affects its axonal function(s). Genome wide CRISPR/Cas9 screens using flow cytometry and optical methods will be undertaken to identify factors that control stathmin-2 synthesis or accumulation. Finally, we will determine the consequences in mice of reduction or loss of stathmin-2 on motor neuron function and muscle innervation/denervation and whether reduction in stathmin-2 synergizes with TDP-43 mutation to drive motor neuron disease. Outcomes of these efforts will provide key insights for understanding basic aspects of axonal and synaptic neurobiology and for evaluating whether maintaining or restoring stathmin-2 is an attractive therapeutic option in sporadic ALS/FTD and ALS from its most frequent genetic cause, repeat expansion in C9orf72.

在受影响的神经元中发现了RNA/DNA结合蛋白TDP-43的细胞质蛋白积累