Modulation of astrocyte-mediated neurotoxicity by NR1D1 in ALS models

ALS 模型中 NR1D1 调节星形胶质细胞介导的神经毒性

基本信息

批准号：
10800001
负责人：
Mariana Atina Pehar
金额：
$ 53.78万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-15 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10800001
关键词：
ALS patients Adult Agonist Amyotrophic Lateral Sclerosis Astrocytes Autopsy Brain Stem Cause of Death Cell Culture Techniques Cells Central Nervous System Circadian Rhythms Coculture Techniques Data Disease Disease Progression Down-Regulation Fibroblasts Functional disorder Genes Goals Homeostasis Human Inflammation Inflammatory Inflammatory Response Link Mammals Mediating Metabolic Metabolism Mitochondria Modeling Molecular Motor Cortex Motor Neuron Disease Motor Neurons Movement Mus Muscle Weakness NF-kappa B Nerve Degeneration Neuroglia Neurons Nuclear Receptors Paralysed Pathologic Pathway interactions Phenotype Play Process Proteins Proteomics Regulation Reporting Role Signal Transduction Skeletal Muscle Specificity Spinal Cord Symptoms Testing Therapeutic Tissues Toxic effect Transcription Repressor Transgenic Organisms adeno-associated viral vector cell type defined contribution familial amyotrophic lateral sclerosis flexibility fused in sarcoma gene therapy glucose metabolism in vivo induced pluripotent stem cell insight knock-down lipid metabolism member metabolomics mitochondrial dysfunction molecular clock motor neuron degeneration mouse model multiple omics mutant nerve stem cell neuroinflammation neuron loss neuronal survival neurotoxic neurotoxicity new therapeutic target novel novel therapeutic intervention overexpression pharmacologic pre-clinical small molecule superoxide dismutase 1 transcriptomics

项目摘要

Abstract ALS or Lou Gehrig's disease is the most common adult-onset motor neuron disease, characterized by the progressive degeneration of motor neurons in the spinal cord, brain stem, and motor cortex. Motor neuron death leads to muscle weakness and paralysis, usually causing death in one to five years after symptoms onset. The molecular mechanisms responsible for motor neuron degeneration in ALS remain uncertain. However, several lines of evidence indicate that dysfunction of glial cells significantly contributes to the neurodegenerative process. Astrocytes, key regulators of central nervous system homeostasis, have been shown to play a major role in the progression of the disease. Accordingly, in cell culture models, astrocytes isolated from ALS mouse models or astrocytes differentiated from fibroblast-derived induced pluripotent stem cells (iPSCs) from sporadic and familial ALS patients, induce motor neuron death. Astrocytes have a key role in the metabolic support of neurons, and lack of metabolic flexibility and mitochondrial dysfunction are hallmarks of ALS astrocytes. We recently showed that NR1D1 plays a critical role in the regulation of astrocyte-motor neuron interaction. NR1D1 is a member of the nuclear receptor superfamily and it is an essential component of the molecular clock in mammals. NR1D1 is also a known repressor of metabolic and inflammatory genes, and has been shown to regulate mitochondrial number and function, glucose and lipid metabolism, and inflammation during normal and pathological conditions. We recently reported that NR1D1 expression decreases in the spinal cord of symptomatic ALS mice. Moreover, we showed that the knockdown of Nr1d1 expression in primary mouse spinal cord astrocyte cultures, or in iPSC- derived human astrocytes, activates NF-kB signaling and induces a pro-inflammatory phenotype that is detrimental for the survival of co-cultured motor neurons. In addition, our preliminary data shows that iPSC- derived astrocytes from ALS patients display lower levels of NR1D1 expression when compared to astrocytes derived from healthy controls. In this proposal, we will investigate the role of NR1D1 in the regulation of astrocyte- motor neuron interaction and the therapeutic potential of targeting NR1D1 in ALS. We will perform an unbiased multi-omic analysis to outline the mechanism by which NR1D1 downregulation induces a neurotoxic phenotype in human iPSC-derived astrocytes (Aim 1). In addition, we will use different approaches to evaluate the effect of modulating NR1D1 expression on the progression of the disease in an ALS mouse model (Aims 2 and 3). The results obtained will contribute to the current understanding of the mechanisms involved in the toxicity of astrocytes towards motor neurons in ALS. In addition, this proposal will establish the therapeutic value of targeting NR1D1 to slow or halt the progression of the disease in an ALS mouse model.

抽象的 ALS 或 Lou Gehrig 病是最常见的成人发病的运动神经元疾病，其特征是脊髓、脑干和运动皮层运动神经元进行性退化。运动神经元死亡导致肌肉无力和瘫痪，通常会在症状出现后一到五年内导致死亡。这导致 ALS 运动神经元变性的分子机制仍不确定。然而，几个一系列证据表明，神经胶质细胞的功能障碍对神经退行性过程有显着影响。星形胶质细胞是中枢神经系统稳态的关键调节因子，已被证明在疾病的进展。因此，在细胞培养模型中，从 ALS 小鼠模型中分离出的星形胶质细胞或星形胶质细胞由散发性和家族性成纤维细胞衍生的诱导多能干细胞 (iPSC) 分化而来 ALS患者，诱发运动神经元死亡。星形胶质细胞在神经元的代谢支持中起着关键作用，并且缺乏代谢灵活性和线粒体功能障碍是 ALS 星形胶质细胞的标志。我们最近展示了 NR1D1 在星形胶质细胞与运动神经元相互作用的调节中起着至关重要的作用。 NR1D1 是以下组织的成员核受体超家族，它是哺乳动物分子钟的重要组成部分。 NR1D1 是也是一种已知的代谢和炎症基因抑制因子，并已被证明可以调节线粒体正常和病理条件下的数量和功能、葡萄糖和脂质代谢以及炎症。我们最近报道，有症状的 ALS 小鼠的脊髓中 NR1D1 表达降低。而且，我们发现，原代小鼠脊髓星形胶质细胞培养物或 iPSC 中 Nr1d1 表达的敲低- 衍生的人星形胶质细胞，激活 NF-kB 信号传导并诱导促炎表型不利于共培养运动神经元的存活。此外，我们的初步数据显示 iPSC- 与星形胶质细胞相比，来自 ALS 患者的星形胶质细胞显示出较低水平的 NR1D1 表达源自健康对照。在本提案中，我们将研究 NR1D1 在星形胶质细胞调节中的作用运动神经元相互作用以及针对 ALS 中 NR1D1 的治疗潜力。我们将公正地执行多组学分析概述 NR1D1 下调诱导神经毒性表型的机制在人 iPSC 衍生的星形胶质细胞中（目标 1）。此外，我们将采用不同的方法来评估效果在 ALS 小鼠模型中调节 NR1D1 表达对疾病进展的影响（目标 2 和 3）。这获得的结果将有助于目前对毒性机制的理解 ALS 中星形胶质细胞朝向运动神经元。此外，该提案将确定以下药物的治疗价值：在 ALS 小鼠模型中，靶向 NR1D1 可减缓或阻止疾病的进展。