RAD23 Control of ALS phenotypes

RAD23 对 ALS 表型的控制

• 批准号: 10274489
• 负责人: Robert G Kalb
• 金额: $ 39.88万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10274489
• 关键词: ALS patients; Ablation; Amyotrophic Lateral Sclerosis; Antisense Oligonucleotides; Autophagocytosis; Behavior; Biochemical; Biochemistry; Biological Models; Caenorhabditis elegans; Cell physiology; Cells; Clinical; Degradation Pathway; Disease; Disease Progression; Equilibrium; Genes; Genetic; Goals; Health; Health Promotion; Histology; Hypersensitivity; Image; Impairment; In Vitro; MG132; Mammals; Mediating; Modeling; Mus; Mutate; Nematoda; Neurodegenerative Disorders; Organism; Pathway interactions; Phenotype; Play; Process; Protein Biosynthesis; Protein Isoforms; Proteins; RAD23B gene; Role; System; Technology; Testing; Therapeutic; Transgenic Mice; Translating; Translations; Ubiquitin; Work; Yeasts; base; experimental study; improved; inhibitor/antagonist; innovation; knock-down; misfolded protein; mouse model; multicatalytic endopeptidase complex; mutant; operation; polyglutamine; protein TDP-43; protein aggregation; protein degradation; proteostasis; tissue culture; ultraviolet irradiation

Abstract The accumulation of damaged, misfolded and aggregation-prone proteins in neurodegenerative diseases reflects corruption of cellular protein homeostasis (or “proteostasis”). Proteostasis is the proper balance of protein synthesis and protein degradation that is required for optimal cellular functioning. The identification of misfolded proteins and their targeting to the major degradative pathways (i.e., the proteasome or the autophagy pathway) are highly regulated processes. A key protein in this process is RAD23. RAD23 promotes the degradation of some proteins and stabilizes cellular levels of other proteins. The mechanism underlying these diametrically opposed operations is not understood. Interestingly, ablation of rad23 accelerates the destruction several disease-causing, aggregation-prone mutated proteins (i.e., polyQ expanded Ataxin3, TDP43, SOD) and confers benefits in a variety of model systems. We hypothesize that RAD23 bridges ubiquitinated misfolded proteins with the proteasome and in doing so, sterically or allosterically inhibits proteasome function. In this way, RAD23 impairs proteostasis. In specific aim #1, we will use imaging and biochemical approaches to test this mechanism of RAD23 action. In in vitro and C.elegans models of ALS, loss of RAD23 is health promoting – whether this is true in mouse models is unknown. Mammals have 2 rad23 isoforms, rad23A and rad23B. In specific aims 2-4, we will use genetic and anti-sense oligomer technology to ablate and/or knockdown rad23A, rad23B or both in several mouse models of ALS. We will comprehensively interrogate the effects of loss of rad23A/B on mouse survival, behavior and biochemistry. The successful completion of these studies has the potential to be translated into clinical therapeutics.

抽象的 受损、错误折叠和易于聚集的蛋白质的积累 神经退行性疾病反映了细胞蛋白质稳态的破坏（或 “蛋白质稳态”）。蛋白质稳态是蛋白质合成和蛋白质降解的适当平衡 这是最佳细胞功能所必需的。错误折叠蛋白的鉴定及其作用 针对主要降解途径（即蛋白酶体或自噬途径） 是受到严格监管的流程。该过程中的一个关键蛋白质是 RAD23。 RAD23 促进 降解一些蛋白质并稳定其他蛋白质的细胞水平。机制 这些截然相反的操作背后的原因尚不清楚。有趣的是，消融 rad23 加速破坏几种致病的、易于聚集的突变蛋白 （即，polyQ 扩展了 Ataxin3、TDP43、SOD）并在多种模型中带来益处 系统。我们假设 RAD23 将泛素化错误折叠蛋白与 蛋白酶体，并在此过程中空间或变构地抑制蛋白酶体功能。这样， RAD23 损害蛋白质稳态。在具体目标#1中，我们将使用成像和生化技术 测试 RAD23 作用机制的方法。在 ALS 的体外模型和线虫模型中， RAD23 的缺失有利于健康——这在小鼠模型中是否属实尚不清楚。 哺乳动物有 2 种 rad23 亚型：rad23A 和 rad23B。在具体目标2-4中，我们将使用遗传 和反义寡聚体技术，以消除和/或敲低 rad23A、rad23B 或两者 几种 ALS 小鼠模型。我们将全面审视损失的影响 rad23A/B 对小鼠生存、行为和生物化学的影响。这些工作的顺利完成 研究有可能转化为临床治疗。