Determination of cell death pathways activated in ALS

摘要/摘要 肌萎缩侧索硬化症(ALS)是引起肌肉的运动神经元障碍的最常见形式 虚弱，残疾，最终死亡。肌萎缩侧索硬化症是由脆弱的上下部细胞选择性死亡引起的 运动神经元。家族性肌萎缩侧索硬化症患者的遗传分析发现了几个基因突变与 ALS，如C9ORF72、TDP-43和SOD1。然而，目前尚不清楚这些突变是如何导致 肌萎缩侧索硬化症患者运动神经元的选择性细胞死亡，以及哪些细胞死亡途径导致 运动神经元细胞死亡。早期的研究表明与细胞凋亡有关，但最近的一项研究 在一个细胞模型中发现了坏死性下垂的作用。多条细胞死亡途径可能被激活。 疾病的病程。我们最近报道了谷胱甘肽过氧化物酶4(Gpx4)的抑制，以及 随后过氧化脂质的积累，触发了一种独特的调节性细胞死亡形式，称为铁下垂。 有趣的是，来自多个肌萎缩侧索硬化症模型的数据以及我们自己的结果表明，铁下垂可能 参与肌萎缩侧索硬化症运动神经元的死亡。基于实验结果表明两者的作用 ALS中的凋亡和非凋亡性细胞死亡途径，我建议检验多个细胞 死亡通路在ALS运动神经元中被激活，集中在铁性下垂的参与。我们的长期合作 目的是阐明细胞死亡通路在人类疾病中是如何调节(和解除调节)的，以及 通过调节细胞死亡途径，促进治疗策略的发展。我们特别是 对了解铁链细胞死亡途径感兴趣。这项提案的目标是确定 肌萎缩侧索硬化症(ALS)运动神经元变性激活的细胞死亡通路 铁性下垂的累及。根据初步数据，该项目将以两个具体目标进行。 在目标1中，我们将使用运动神经元系(iPS-MNS)识别与ALS相关的特定细胞死亡途径 在体外和体内使用ALS转基因小鼠模型。在目标2中，我们将确定 铁下垂抑制剂在ALS iPS-MN细胞系和SOD1-G93A转基因小鼠中的单药或AS 与其他细胞死亡方式的抑制物联合治疗。这项研究将有助于揭示 肌萎缩侧索硬化症运动神经元变性的分子机制，以及铁性下垂在疾病中的作用。它 也可能导致新的治疗策略的发展。此外，这个Score-2奖将增强 圣约翰大学的研究环境，并为积极进取的学生提供了许多机会 从不同的来源学习生物医学研究的基础知识。

SUMMARY/ABSTRACT Amyotrophic lateral sclerosis (ALS) is the most common form of motor neuron disorder that causes muscle weakness, disability, and eventually death. ALS is caused by selective cell death of vulnerable upper and lower motor neurons. Genetic analyses in familial ALS patients have identified several genes with mutations linked to ALS, such as C9ORF72, TDP-43, and SOD1. However, it remains unclear how these mutations lead to selective cell death of motor neurons in ALS patients, and which cell death pathways are responsible for the motor neuron cell death. Early studies suggested the involvement of apoptosis, but a more recent investigation found a role for necroptosis in one cell model. It is plausible that multiple cell death pathways are activated in the course of the disease. We recently reported that inhibition of glutathione peroxidase 4 (GPX4), and subsequent accumulation of lipid peroxides, triggers a unique form of regulated cell death termed ferroptosis. Intriguingly, data from multiple ALS models as well as our own results have suggested that ferroptosis may contribute to the motor neuron cell death in ALS. Based on the experimental results indicating the role of both apoptotic and non-apoptotic cell death pathways in ALS, I propose to test the hypothesis that multiple cell death pathways are activated in ALS motor neurons, focusing on the involvement of ferroptosis. Our long-term goal is to elucidate how cell death pathways are regulated (and deregulated) in human disease, and to contribute to the development of treatment strategies by modulating cell death pathways. We are particularly interested in understanding the ferroptotic cell death pathway. The objective of this proposal is to determine cell death pathways activated in degenerating motor neurons in amyotrophic lateral sclerosis (ALS) focused on the involvement of ferroptosis. Based on the preliminary data, the project will be pursued in two specific aims. In Aim 1, we will identify the specific cell death pathways involved in ALS using motor neuron lines (iPS-MNs) in vitro and using a transgenic mouse model of ALS in vivo. In Aim 2, we will determine the efficacy of ferroptosis inhibitor in ALS iPS-MN cell lines and in SOD1-G93A transgenic mice either as a single-agent or as a combination therapy with suppressors of other cell death modalities. This research will shed light on the molecular mechanism of motor neuron degeneration in ALS, as well as the role of ferroptosis in the disease. It may also lead to the development of new therapeutic strategies. In addition, this SCORE-2 award will enhance the research environment at St. John's University and provide numerous opportunities for motivated students from diverse origin to learn the fundamentals of biomedical research.