Defining the altered FUS-PARP-1-DNA Ligase III axis and its implications to nuclear and mitochondrial genome damage response in Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Dementia (FTD)

定义改变的 FUS-PARP-1-DNA 连接酶 III 轴及其对肌萎缩侧索硬化症 (ALS) 和额颞叶痴呆 (FTD) 中核和线粒体基因组损伤反应的影响

• 批准号: 9980670
• 负责人: Muralidhar L Hegde
• 金额: $ 201.88万
• 依托单位: METHODIST HOSPITAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9980670
• 关键词: ALS patients; Affect; Amyotrophic Lateral Sclerosis; Architecture; Astrocytes; Basic Science; CRISPR/Cas technology; Cells; Cessation of life; Clinical; Communication; Complex; DNA; DNA Damage; DNA Double Strand Break; DNA Ligases; DNA Ligation; DNA Repair; DNA Single Strand Break; DNA ligase III; DNA strand break; DNA-Binding Proteins; Data; Defect; Disease Progression; Double Strand Break Repair; Down-Regulation; Energy Metabolism; Etiology; Event; Familial Amyotrophic Lateral Sclerosis; Follow-Up Studies; Frontotemporal Dementia; Genes; Genome; Genome Stability; Genomic Instability; Goals; Hospitals; Human; In Vitro; Individual; Intervention; Knock-in; Lead; Ligase; Ligation; Light; Link; Maintenance; Mediating; Methodist Church; Mitochondria; Molecular; Motor Neurons; Muscular Atrophy; Mutation; NADH; Nature; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nuclear; Oxidative Stress; Oxides; Pathologic; Pathology; Pathway interactions; Patients; Physiological; Point Mutation; Proteins; Publications; RNA; Reactive Oxygen Species; Recipe; Research; Research Institute; Resources; Role; Signal Transduction; Site; Spinal Cord; Stress; Symptoms; Techniques; Testing; Tissues; Toxic effect; Transgenic Organisms; Translational Research; Untranslated RNA; XRCC1 gene; autosomal dominant mutation; brain tissue; effective therapy; follow-up; frontal lobe; frontotemporal lobar dementia-amyotrophic lateral sclerosis; genome integrity; induced pluripotent stem cell; innovation; insight; loss of function; member; mitochondrial dysfunction; mitochondrial genome; mouse model; mutant; mutation correction; nerve stem cell; neuron loss; neuroprotection; neurotoxicity; novel; oxidative damage; recruit; repaired; response; sarcoma; senescence; stress granule; targeted treatment; therapeutic target; transcriptome sequencing; translational impact

Genome damage and defective repair are etiologically linked to Fused in Sarcoma (FUS)-associated amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). However, the underlying mechanisms remain enigmatic, which is a roadblock for exploiting genome repair-targeted therapies for ALS/FTD. Our recent publication (Wang et al, Nature Communications, 2018) identified defects in DNA nick ligation and oxidative damage repair in a subset of ALS patients, caused by mutations in the RNA/DNA-binding protein FUS. In healthy neurons, FUS protects the genome by facilitating PARP1-dependent recruitment of XRCC1/DNA Ligase IIIα (LigIII) to oxidized genome sites and activates LigIII via direct interaction. We discovered that FUS toxicity caused significantly decreased recruitment of XRCC1/LigIII to DNA strand breaks. DNA ligation defects in ALS patient-derived iPSC lines carrying FUS mutations and in subsequently generated motor neurons were rescued by CRISPR/Cas9-mediated mutation correction. Moreover, our follow-up studies showed substantially reduced auto and total PARylation activity of PARP-1 both in vitro and in cell, after loss of FUS or mutant expression, which in addition to regulating LigIII/XRCC1 recruitment at damage sites, could impact neuronal energy metabolism by uncoupling NAD+/NADH levels and stress granule dynamics in motor neurons. Collectively these events may provide a recipe for neurodegeneration. These findings that uncovered a new pathway of defective DNA ligation and PARP-1 functions in FUS-linked ALS-FTD, raised three key questions that need to be investigated to understand their implications in neuronal death and to develop a comprehensive strategy of PARylation and LigIII targeted interventions for ameliorating FUS-associated ALS-FTD. These questions are: (1) How does FUS affect PARP-1's PARylation activity and what is its impact on genome maintenance and energy metabolism? (2) What is the effect of FUS-mediated LigIII inhibition on the mitochondrial genome and its functions? This is important as LigIII is the only DNA ligase for both replication and repair in mitochondria, and both FUS and PARP-1 localize in mitochondria. (3) What is the effect of FUS pathology on microhomology-mediated alternative end-joining (MMEJ) pathway of DNA double strand break repair, which involves LigIII, XRCC1 and PARP-1?. This project, will utilize human patient-derived iPSC lines harboring FUS mutations, their isogenic controls with mutation correction by CRISPR/Cas9 knock-in strategy, a transgenic FUS-∆NLS mouse model and human ALS, FTD patient spinal cord/brain tissue, to test our novel hypothesis that FUS pathology-mediated DNA ligation defects via reduced PARylation inhibits oxidative genome damage repair and promotes neurodegeneration. We will further show that rescuing Ligase and PARP functions are promising avenues for neuroprotection. Our studies investigating the previously unexplored link between altered FUS-PARP-1-LigIII axis and ALS-FTD are both technically and conceptually innovative, have important immediate and long term goals and will strongly impact translational ALS-FTD research.

基因组损伤和叛逃者损害在病因上与肉瘤（FUS）相关的融合有关 肌萎缩性侧面硬化症（ALS）和额颞痴呆（FTD） 保持神秘性，这是我们最近的ALS/FTD爆炸基因组修复疗法的障碍。 出版物（Wang等人，自然通讯，2018年）确定了DNA结扎和氧化中的缺陷 由RNA/DNA结合蛋白FUS中的突变引起的ALS特定性的损伤修复 神经元，FUS通过促进XRCC1/DNA连接酶IIIα的PARP1依赖性募集来保护基因组 （ligiii）氧化的基因组位点并通过直接相互作用激活利格。 XRCC1/Ligiii的募集显着降低到DNA链中，随后产生的运动神经元被救出。 CRISPR/CAS9介导的突变校正。 在体外和细胞中，PARP-1的总核化活性，在大惊小怪或突变体express之后， 除了调节ligiii/XRCC1在损害处募集的外，还可能影响神经元能量代谢 将运动神经元中的NAD+/NADH水平和应力颗粒动力学取消耦合。 提供神经变性的配方。 和PARP-1在FUS链接的ALS-FTD中起作用，raiss提出了三个关键问题，需要调查 了解神经元死亡的含义，并制定划界的全面策略 ligiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiii。 影响PARP-1的抚养活性，其对基因组维持和能量代谢的影响是什么？ FUS介导的LiGIII抑制对线粒体基因组及其功能的影响是什么？ 重要的是，因为Ligiii是线粒体复制和修复的唯一DNA连接酶，以及Futh Fus Fus和 PARP-1位于线粒体中。 DNA双重分散休息修复的最终连接（MMEJ）途径，其中涉及LiGIII，XRCC1和PARP-1？ 该项目将利用人类衍生的IPSC林斯林斯（Linsc Linsc 通过CRIS/CAS9敲门策略，转基因FUS-NLS Mousel和人类ALS的突变校正， FTD患者脊髓/脑组织 通过降低的核化缺陷抑制氧化基因组损害，一些促进 神经变性。 对于神经保护作用。 术语目标和将Stranslatitionals-FTD研究。

项目成果

Mitochondria-Targeted Oligomeric α-Synuclein Induces TOM40 Degradation and Mitochondrial Dysfunction in Parkinson's Disease and Parkinsonism-Dementia of Guam.

线粒体靶向寡聚 α-突触核蛋白在关岛帕金森病和帕金森痴呆症中诱导 TOM40 降解和线粒体功能障碍。

• DOI: 10.21203/rs.3.rs-3970470/v1
• 发表时间: 2024
• 期刊: Research square
• 作者: Hegde,Muralidhar;Vasquez,Velmarini;Kodavati,Manohar;Mitra,Joy;Vendula,Indira;Hamilton,Dale;Garruto,Ralph;Rao,KS
• 通讯作者: Rao,KS