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Defining the mechanisms and consequences of nuclear defects in ALS/FTD

定义 ALS/FTD 核缺陷的机制和后果

基本信息

批准号：
10599867
负责人：
Claudia Fallini
金额：
$ 38.01万
依托单位：
UNIVERSITY OF RHODE ISLAND
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：
2021-04-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10599867
关键词：
Actins Adult Affect Aging Binding C9ORF72 CREB1 gene Cell Nucleus Cell model Cells Cytoplasm Cytoskeleton DNA DNA Binding DNA-Binding Proteins Defect Dendrites Development Disease Disease Progression Failure Functional disorder Gene Expression Gene Expression Profile Genes Genetic Genetic Transcription Homeostasis Human Impaired cognition Impairment Induced pluripotent stem cell derived neurons Investigation Knowledge Lead Link Messenger RNA Modeling Molecular Morphology Motor Neurons Muscle Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Nuclear Nuclear Import Nuclear Lamina Nuclear Pore Complex Outcome Paralysed Pathogenesis Pathogenicity Pathology Pathway Analysis Pathway interactions Phenotype Physiological Play Population Process Proteins Proteome Proteomics RNA RNA Binding RNA Splicing RNA-Binding Proteins Regulation Research Role Spliced Genes Stress and Coping Structure Synapses Testing Therapeutic Traction Transcriptional Activation Transcriptional Regulation Translations axonal degeneration cellular pathology coping design familial amyotrophic lateral sclerosis frontal lobe frontotemporal lobar dementia amyotrophic lateral sclerosis genetic approach innovation insight neuron loss neuronal survival novel novel therapeutic intervention nucleocytoplasmic transport pharmacologic postmitotic protein TDP-43 reduce symptoms resilience response therapeutically effective transcription factor transcriptome sequencing transcriptomics

项目摘要

PROJECT SUMMARY A detailed insight into the molecular defects leading to neuronal death in ALS/FTD is still missing, hindering the development of a cure. Defects in the nuclear pore complex (NPC) and nucleocytoplasmic transport (NCT) have been proposed to play a central role in ALS/FTD pathology. An outstanding knowledge gap is what are the mechanisms, consequences, and pathogenic relevance of impaired NCT on neuronal resilience in neurodegenerative diseases and during aging. The objective of this application is to identify at the mechanistic level the novel cellular and molecular pathways that are dysfunctional and cause the failure of the NCT ultimately leading to neurodegeneration. In particular, we will test the hypothesis that the actin cytoskeleton is a main modulator of nuclear stability and NPC function, and that changes to this pathway in ALS/FTD lead to a diminished ability of neurons to cope with stress and physiological stimulation. The specific aims of this proposal are to: 1) Define how actin regulates the function and dysfunction of the NPC. We will perform a systematic and in-depth investigation of the molecular interactions between actin and the NPC using pharmacological and genetic approaches in iPSC-derived neurons carrying the C9ORF72 mutation. 2) Identify the consequences of NCT defects on the nuclear and cytoplasmic proteome in response to cellular stimulation. By combining an “omics” approach with a candidate approach (i.e. CREB pathway), we will analyze changes in the nuclear and cytoplasmic proteome in iPSC-derived neuronal models of ALS/FTD following stimulations, and we will test the potentials of cytoskeleton modulation to rescue such defects. 3) Define how defects in nuclear import affect the transcriptional response to cellular stimulation. We will analyze changes in gene expression and splicing profile in ALS/FTD neurons following cell stimulation by RNA-Seq. Network analysis performed by integrating the results from the transcriptomic and proteomics approaches will identify ALS/FTD relevant altered pathways. Modulation of the cytoskeleton will be used to rescue the identified defects. At the completion of this research, we will have identified the molecular and cellular mechanisms that control the stability and function of the NPC, and the functional consequences that the disruption of NCT has on the resilience of neurons in ALS/FTD. Gaining insights on what pathways upstream and downstream of the NPC are affected in ALS/FTD will greatly expand our understanding of disease pathogenesis and will allow us to identify yet unexplored avenues for therapy in these diseases. Ultimately, we expect that our research will lead to more effective therapeutic strategies that take advantage of the crosstalk between NPC, RNA regulation, and cytoskeleton. This research will have a broad impact on a spectrum of diseases including but not limited to ALS/FTD.

项目概要对导致 ALS/FTD 神经元死亡的分子缺陷的详细了解仍然缺乏，这阻碍了治疗方法的开发。核孔复合体 (NPC) 和核质转运 (NCT) 缺陷已被认为在 ALS/FTD 病理学中发挥核心作用。突出的知识差距是什么 NCT 受损对神经元弹性的机制、后果和致病相关性神经退行性疾病和衰老过程中。该应用程序的目的是识别机械平衡功能失调并导致 NCT 失败的新细胞和分子途径最终导致神经退行性变。特别是，我们将检验以下假设：肌动蛋白细胞骨架是核稳定性和 NPC 功能的主要调节剂，并且 ALS/FTD 中该途径的改变导致神经元应对压力和生理刺激的能力下降。该提案的具体目标是：1）定义肌动蛋白如何调节肌动蛋白的功能和功能障碍。全国人大。我们将对肌动蛋白和肌动蛋白之间的分子相互作用进行系统和深入的研究 NPC 在携带 C9ORF72 的 iPSC 衍生神经元中使用药理学和遗传学方法突变。 2) 确定NCT缺陷对细胞核和细胞质蛋白质组的影响对细胞刺激的反应。通过将“组学”方法与候选方法（即 CREB 途径），我们将分析 iPSC 衍生神经元模型中核和细胞质蛋白质组的变化刺激后 ALS/FTD 的发生率，我们将测试细胞骨架调节的潜力，以挽救这种情况缺陷。 3) 定义核输入缺陷如何影响细胞的转录反应刺激。我们将分析 ALS/FTD 神经元基因表达和剪接特征的变化，如下 RNA-Seq 的细胞刺激。通过整合转录组和转录组的结果进行网络分析蛋白质组学方法将识别 ALS/FTD 相关的改变途径。细胞骨架的调节将是用于挽救已识别的缺陷。这项研究完成后，我们将确定控制的分子和细胞机制 NPC 的稳定性和功能，以及 NCT 的破坏对 NPC 的功能影响 ALS/FTD 中神经元的恢复能力。深入了解 NPC 的上游和下游路径受 ALS/FTD 影响的患者将极大地扩展我们对疾病发病机制的理解，并使我们能够确定这些疾病的尚未探索的治疗途径。最终，我们期望我们的研究将引领更有效的治疗策略，利用 NPC、RNA 调节之间的串扰，和细胞骨架。这项研究将对一系列疾病产生广泛影响，包括但不限于 ALS/FTD。