Roles for TDP-43 and FUS in ALS Using Motor Neurons from Embryonic Stem Cells

TDP-43 和 FUS 在使用胚胎干细胞运动神经元的 ALS 中的作用

• 批准号: 8312031
• 负责人: Dara Ditsworth
• 金额: $ 5.39万
• 依托单位: LUDWIG INSTITUTE FOR CANCER RES LTD
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8312031
• 关键词: Adult; Affect; Amyotrophic Lateral Sclerosis; Biology; Brain; Cell Line; Cells; Complementary DNA; DNA; DNA-Binding Proteins; Disease; Disease Progression; ES Cell Line; Future; Gene Expression Profile; Genes; Goals; Human; Inherited; Lead; Light; Motor Neurons; Movement; Mus; Muscle; Mutation; Neurodegenerative Disorders; Neuroglia; Neurons; Onset of illness; Paralysed; Pathogenesis; Pathology; Pathway interactions; Patients; Peptides; Phenotype; Prions; Proliferating; Property; Proteins; RNA; RNA Processing; RNA-Binding Proteins; Role; Signal Pathway; Small Interfering RNA; Spinal Cord; Tetracyclines; Therapeutic; Toxic effect; Transgenic Mice; Weight; base; cell type; embryonic stem cell; gain of function; loss of function; mutant; nervous system disorder; premature; promoter; protein TDP-43; protein function; sarcoma; small hairpin RNA; success; therapeutic development

DESCRIPTION (provided by applicant): Amyotrophic lateral sclerosis (ALS) is an adult-onset neurodegenerative disorder in which premature loss of upper and lower motor neurons leads to fatal paralysis with a typical disease course of one to five years. Mutations in two DNA/RNA binding proteins, TAR DNA-binding protein (TDP-43) and Fused in Sarcoma (FUS) have recently been identified as primary causes of inherited ALS and have led to what is likely to be a paradigm shift in efforts to understand the pathogenesis of ALS. Yet there remains a need to examine the functions of these proteins in cell types relevant for ALS progression, both motor neurons and glial cells. While a contribution from surrounding cells is well established in inherited ALS caused by SOD1 mutations, the impact of non-cell autonomous mechanisms in disease progression following mutation in TDP-43 or FUS is completely unknown. This proposal seeks to use embryonic stem cells derived from existing transgenic mice to examine the consequences of either reduction or mutation of TDP-43 or FUS in purified motor neurons. Cell intrinsic consequences of alterations in these genes will be determined by inducing differentiation of embryonic stem cells into motor neurons which either lack or express mutant TDP-43 or FUS. Rather than take a candidate approach to pursue the few genes that have already been identified as targets of TDP-43 or FUS in proliferating cell lines, isolated motor neuron cultures will be used to identify the complete set of RNAs that are altered by loss of TDP-43 or FUS in Aim 1, and then in Aim 2 to ask whether any RNAs are altered upon mutation of TDP-43 or FUS and if so, if they are the same as those observed upon loss of function. By taking such a comprehensive and systematic approach from purified starting material it is likely that the specific targets and signaling pathways affected can be uncovered - discoveries that might elucidate underlying mechanisms for disease and provide a basis for future therapeutic developments. Success in these goals will lead to greater understanding of ALS disease mechanism and will provide rationale for future studies in glial cell types to determine whether manipulation of supporting non-neuronal cells would provide therapeutic benefit in ALS patients with mutations in TDP-43 or FUS.

描述（由申请人提供）：肌萎缩侧索硬化症（ALS）是一种成人发病的神经退行性疾病，其中上运动神经元和下运动神经元过早丧失导致致命性瘫痪，典型病程为一到五年。两种 DNA/RNA 结合蛋白 TAR DNA 结合蛋白 (TDP-43) 和肉瘤融合蛋白 (FUS) 的突变最近被确定为遗传性 ALS 的主要原因，并可能导致理解 ALS 发病机制的工作发生范式转变。然而，仍然需要检查这些蛋白质在与 ALS 进展相关的细胞类型（运动神经元和神经胶质细胞）中的功能。虽然周围细胞对 SOD1 突变引起的遗传性 ALS 的影响已得到充分证实，但非细胞自主机制对 TDP-43 或 FUS 突变后疾病进展的影响完全未知。 该提案旨在使用源自现有转基因小鼠的胚胎干细胞来检查纯化的运动神经元中 TDP-43 或 FUS 减少或突变的后果。这些基因改变的细胞内在后果将通过诱导胚胎干细胞分化为缺乏或表达突变TDP-43或FUS的运动神经元来确定。不是采用候选方法来追踪增殖细胞系中已被确定为 TDP-43 或 FUS 靶标的少数基因，而是使用分离的运动神经元培养物来鉴定在目标 1 中因 TDP-43 或 FUS 丢失而改变的完整 RNA 集，然后在目标 2 中询问是否有任何 RNA 在 TDP-43 或 FUS 突变时发生改变，如果是，它们是否相同 正如在功能丧失时观察到的那样。 通过从纯化的起始材料中采取这种全面、系统的方法，可能会发现受影响的特定靶点和信号通路——这些发现可能会阐明疾病的潜在机制，并为未来的治疗发展提供基础。这些目标的成功将导致人们更好地了解 ALS 疾病机制，并为未来对神经胶质细胞类型的研究提供理论基础，以确定支持性非神经元细胞的操作是否能为 TDP-43 或 FUS 突变的 ALS 患者提供治疗益处。