Profiles of Common and Unique aspects of Upper Motor Neuron degeneration in HSP and ALS

HSP 和 ALS 上运动神经元变性的共同和独特方面的概况

• 批准号: 10526893
• 负责人: Pembe Hande Ozdinler
• 金额: $ 44万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10526893
• 关键词: ALS pathology; ALS patients; Actins; Amyotrophic Lateral Sclerosis; Attention; Binding Proteins; Biology; Brain; Cell physiology; Characteristics; Clinical; Complex; Defect; Disease; Event; Experimental Designs; Fluorescence; Foundations; Future; Gene Expression; Gene Expression Profile; Genes; Genetic; Genotype; Goals; Hereditary Spastic Paraplegia; Human; Investigation; Knowledge; Label; Modeling; Molecular; Molecular Genetics; Motor Cortex; Motor Neuron Disease; Motor Neurons; Movement; Mus; Mutate; Mutation; Nerve Degeneration; Neurons; Pathology; Pathway interactions; Patients; Population; Protein Dynamics; Proteins; Proteomics; RNA Splicing; Reporter; Reverse Transcriptase Polymerase Chain Reaction; Role; Sampling; Structure; Therapeutic; Therapeutic Intervention; Time; Transcript; Tubulin; UCHL1 gene; Upper Motor Neuron Disease; Variant; Western Blotting; cell type; druggable target; effective therapy; immunocytochemistry; insight; interest; internal control; metabolomics; motor neuron degeneration; mouse model; neuron loss; neuropathology; profilin; protein TDP-43; spastin; transcriptome sequencing; treatment strategy

ABSTRACT/ PROJECT SUMMARY: The movement starts in the brain. Especially for motor neuron diseases, which impact voluntary movement, the brain component requires better understanding and assessment. Upper motor neurons (UMNs), located in layer 5 of the motor cortex, have a unique role for the initiation and modulation of voluntary movement. Their progressive degeneration is the characteristic hallmark of neuropathology observed in hereditary spastic paraplegia (HSP) and amyotrophic lateral sclerosis (ALS) patients. In striking contrast to their clinical importance and relevance, very little is known about the underlying causes of their vulnerability and progressive loss. This lack of information and understanding of the cellular and genetic basis of their vulnerability, as well as the common and unique aspects of their neurodegeneration hinders our ability to develop effective and long-term treatment strategies for diseases of the UMN. In an effort to bring a mechanistic insight into the cellular and molecular basis of UMN degeneration in two distinct diseases, such as HSP and ALS, we are going to take advantage of the UCHL1-eGFP mice, a reporter line in which the UMNs are genetically labeled with fluorescence. In this proposal, we will focus our attention to the Profilin and Spastin, because like Spastin, Profilin is an actin/tubulin-binding protein, and mutations in profilin results in ALS, whereas mutations in spastin results in HSP. To date numerous mouse models are generated to mimic the human condition. We crossed the hPFN1G118V (generated by Dr. Kiaei) and SPASTC448Y mice (generated by Dr. Baas) with UCHL1-eGFP mice to generate the UMN reporter lines with these mutations. Since we have shown that the UMNs of ALS patients with TDP-43 pathology and mouse UMNs of TDP-43 model share the same aspects of neurodegeneration at a cellular level, we will focus our attention to the neurons and purify diseased UMNs from the complex structure of the brain as a “pure” neuron population at different stages of the disease to investigate the dynamic changes in their gene expression profile as well as the proteins that are present in them, with respect to the healthy UMNs. As an internal control, we will use UMNs isolated from TDP- 43 mouse model not only because TDP-43 pathology is widely observed in ALS and HSP, but also because this model better represents the spectrum of ALS. Upon completion, our results will begin to reveal the common and unique aspects of UMN vulnerability in HSP and ALS, it will also suggest the distinct set of genes and/or proteins that are responsible for the initiation of their vulnerability. Most importantly our results have the potential to identify druggable targets and pathways of interest for future therapeutic interventions.

摘要/项目摘要： 运动始于大脑。特别是针对运动神经元疾病，影响自愿运动， 大脑成分需要更好的理解和评估。上层运动神经元（UMNS），位于层 运动皮层的5个，在自愿运动的主动性和调制方面具有独特的作用。他们的 渐进式变性是在神经痉挛中观察到的神经病理学的特征标志 截瘫（HSP）和肌萎缩性侧性硬化症（ALS）患者。罢工与他们的临床重要性形成鲜明对比 和相关性，关于其脆弱性和渐进式损失的根本原因知之甚少。这 缺乏信息以及对其脆弱性的细胞和遗传基础的理解，以及 神经退行性的共同和独特的方面阻碍了我们发展有效和长期的能力 UMN疾病的治疗策略。 为了将机械洞察力带入两个不同的umn变性的细胞和分子基础 疾病，例如HSP和ALS，我们将利用UCHL1-EGFP小鼠的优势， UMN在遗传上用荧光标记。在此提案中，我们将把注意力集中在 叶片和氨纶，因为像Spastin一样，profilin是肌动蛋白/微管蛋白结合蛋白，而Profilin中的突变 结果为ALS，而Spastin的突变会导致HSP。迄今为止生成了许多鼠标模型 模仿人类状况。我们越过HPFN1G118V（由Kiaei博士生成）和Spastc448y小鼠 （由BAA博士生成）用UCHL1-EGFP小鼠产生使用这些突变的UMN报告基因。自从 我们已经表明，TDP-43病理学和TDP-43模型共享的TDP-43病理和小鼠UMN的UMNS 神经退行性的相同方面在细胞水平上，我们将注意力集中在神经元上并纯化 大脑复杂结构的患病umn是在不同阶段的“纯”神经元种群 疾病研究其基因表达谱的动态变化以及 就健康的UMN而言存在。作为内部控制，我们将使用从TDP-隔离的UMNS 43小鼠模型不仅是因为TDP-43病理在ALS和HSP中广泛观察到，而且还因为这是因为 模型更好地表示ALS的频谱。 完成后，我们的结果将开始揭示HSP中UMN脆弱性的常见和独特方面 和ALS，它还将提出负责其主动性的独特基因和/或蛋白质集 脆弱性。最重要的是，我们的结果有可能识别可吸毒目标和感兴趣的途径 用于将来的治疗干预措施。