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

• 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
• 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; 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

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氏病是最常见的成年运动神经元疾病，其特征是 脊髓，脑干和运动皮质中运动神经元的逐渐变性。运动神经元死亡 导致肌肉无力和瘫痪，通常在症状发作后一到五年内导致死亡。这 负责ALS运动神经元变性的分子机制仍然不确定。但是，有几个 证据线表明，神经胶质细胞的功能障碍显着有助于神经退行性过程。 星形胶质细胞是中枢神经系统稳态的关键调节剂，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 与成纤维细胞衍生的诱导的多能干细胞（IPSC）不同的星形胶质细胞来自零星和家族性 ALS患者，诱导运动神经元死亡。星形胶质细胞在神经元的代谢支持中具有关键作用，并且 缺乏代谢柔韧性和线粒体功能障碍是ALS星形胶质细胞的标志。我们最近显示 NR1D1在体形胶质运动神经元相互作用的调节中起关键作用。 NR1D1是 核受体超家族，是哺乳动物分子钟的重要组成部分。 NR1D1是 也是已知的代谢和炎症基因的抑制剂，已被证明可以调节线粒体 在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。 我们最近报道说，有症状的ALS小鼠的脊髓中NR1D1的表达降低。而且， 我们表明，原代小鼠脊髓星形胶质细胞培养物或IPSC-中NR1D1表达的敲低 衍生的人类星形胶质细胞，激活NF-KB信号并诱导促炎的表型 不利于共培养运动神经元的生存。此外，我们的初步数据表明IPSC- 与星形胶质细胞相比 源自健康对照。在此提案中，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和靶向ALS中NR1D1的治疗潜力。我们将执行公正 多词分析概述了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（AIM 1）。此外，我们将使用不同的方法来评估 在ALS小鼠模型中调节NR1D1对疾病进展的表达（目标2和3）。这 获得的结果将有助于当前对涉及毒性机制的理解 ALS的运动神经元的星形胶质细胞。此外，该建议将确定 靶向NR1D1在ALS小鼠模型中降低或停止疾病的进展。