Elucidating the impact of FIC-1/FICD-mediated AMPylation on polyglutamine aggregation dynamics and toxicity

阐明 FIC-1/FICD 介导的 AMPylation 对聚谷氨酰胺聚集动力学和毒性的影响

• 批准号: 10464265
• 负责人: Kate Matthys Van Pelt
• 金额: $ 3.95万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10464265
• 关键词: Adenosine Monophosphate; Affect; Aging; Alleles; Animals; Attenuated; Biochemical; Biological Assay; Buffers; CRISPR interference; Caenorhabditis elegans; Cells; Cellular Structures; Chemicals; Data; Development; Disease; Disease model; Excision; Failure; Family; General Population; Genes; Genetic; Goals; Heat-Shock Proteins 70; Human; Huntington Disease; Huntington gene; In Vitro; Inclusion Bodies; Individual; Inherited; Knock-out; Knowledge; Length; Link; MJD1 protein; Machado-Joseph Disease; Maintenance; Mediating; Mediator of activation protein; Molecular; Molecular Chaperones; Mutagenesis; Neurodegenerative Disorders; Neurons; Orthologous Gene; Pathogenicity; Pathologic; Pathway interactions; Patients; Physiological; Post-Translational Protein Processing; Post-Translational Regulation; Prevalence; Process; Proteins; Quality Control; RNA Interference; Regulation; Reporting; Research; Role; Signal Pathway; Solubility; Stress; Sum; Techniques; Testing; Therapeutic; Therapeutic Intervention; Tissues; Toxic effect; Trinucleotide Repeat Expansion; Type 1 Spinocerebellar Ataxia; Work; arm; base; biological adaptation to stress; disease-causing mutation; follow-up; functional decline; human embryonic stem cell; in vitro Model; knock-down; misfolded protein; mutant; neuron loss; novel; overexpression; polyglutamine; protein aggregation; protein function; protein misfolding; proteostasis; response; stem cells; targeted treatment

PROJECT SUMMARY Huntington’s disease (HD) and spinocerebellar ataxia type 3 (SCA3) are inherited aging-associated diseases that have a devastating impact on patients and family caretakers relative to their prevalence in the general population. These conditions belong to a family of polyglutamine (polyQ) expansion diseases caused by mutations resulting in the pathological expansion of trinucleotide (CAGn) repeats in distinct genes. Increases in CAG repeat length give rise to proteins containing aberrantly expanded polyQ tracts, which interfere with normal protein function and promote misfolding. Toxicity in these diseases is thought to arise in part from the formation of pathological inclusion bodies comprised of aberrantly conformed mutant proteins, a hallmark observed in numerous aging-associated neurodegenerative diseases. Despite extensive efforts to decipher the mechanisms underlying toxicity in polyQ diseases, however, little progress has been made towards identifying targets for therapeutic intervention. Recently, the post-translational modification (PTM), AMPylation, has emerged as a novel regulator of HSP70 family chaperones, crucial components of the cell’s protein quality control machinery that buffer against protein misfolding stress. Protein AMPylation is carried out by the fic-type AMPylase, FICD in humans, and its ortholog FIC-1, in C. elegans, respectively. Work in our lab has established that FIC-1-mediated AMPylation directly alters polyQ aggregation dynamics and toxicity. Further, my preliminary data as presented in this proposal identifies fic-1 deficiency as sufficient to rescue survival of C. elegans expressing aggregation-prone polyQs during development in a polyQ length-dependent manner. Taken together, these findings suggest that the loss of FIC- 1/FICD-mediated AMPylation bolsters proteostasis network capacity to alleviate toxicity induced by polyQ protein aggregation. This project will utilize cross-disciplinary approaches to generate a holistic characterization of FICD/FIC-1- mediated AMPylation in polyQ diseases. To this end, I will harness the powerful genetics of C. elegans to uncover novel pathway(s) that promote survival in the face of pathogenic polyQ aggregation (Aim 1). In tandem, I will employ functional assays in neurons derived from HD and SCA3 patient stem cells to profile how FICD activity affects polyQ aggregation and toxicity in these disease models (Aim 2). The results of these studies will advance our knowledge of how AMPylation regulates proteostasis in polyQ diseases. The ultimate goal of my research is to capitalize on these findings to develop translatable therapeutic approaches for aging-associated diseases.

项目总结 亨廷顿病(HD)和脊髓小脑性共济失调3型(SCA3)是遗传性衰老相关疾病 与患者和家庭照顾者的普遍患病率相比，这些疾病对患者和家庭照顾者具有毁灭性的影响 人口。这些疾病属于聚谷氨酰胺(PolyQ)扩张性疾病家族，由 突变导致不同基因中三核苷酸(CAGn)重复序列的病理性扩张。增加了 CAG重复长度导致含有异常扩张的多聚Q链的蛋白质，这干扰了正常 蛋白质的功能和促进错误折叠。这些疾病的毒性被认为部分是由于形成的 病理性包涵体由异常整合的突变蛋白组成，这是在 许多与衰老相关的神经退行性疾病。尽管人们做出了大量努力来破译这些机制 然而，多药耐药疾病的潜在毒性在确定靶点方面进展甚微。 治疗性干预。 最近，翻译后修饰(PTM)，即AMP化，已经成为HSP70的一种新的调节因子 家族伴侣是细胞蛋白质质量控制机制的关键组成部分，可以缓冲蛋白质 错折应力。在人类中，蛋白质AMPylase FICD及其同源基因是通过Fic型AMPylase来执行的 FIC-1，分别在线虫中。我们实验室的工作已经证实，FIC-1介导的AMP化直接改变 多Q聚集动力学和毒性。此外，我在这项提案中提出的初步数据表明 FIC-1缺陷足以挽救表达易于聚集的多QS的线虫在 以依赖于多个Q长度的方式发展。综上所述，这些发现表明，FIC的损失- 1/FICD介导的AMP化增强蛋白稳定网络能力以减轻多聚Q蛋白诱导的毒性 聚合。 该项目将利用跨学科的方法来生成FICD/FIC-1的整体特征- 多聚Q疾病中介导的AMP化。为此，我将利用线虫强大的遗传学来揭示 面对致病多Q聚集促进生存的新途径(S)(目标1)。一齐，我会 利用HD和SCA3患者干细胞来源的神经元的功能分析来描述FICD的活性 在这些疾病模型中影响多Q聚集和毒性(目标2)。这些研究的结果将会取得进展 我们对AMP化如何调节多聚Q疾病中的蛋白平衡的了解。我研究的最终目标是 利用这些发现，开发可翻译的治疗衰老相关疾病的方法。