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

基本信息

批准号：
10656221
负责人：
Kate Matthys Van Pelt
金额：
$ 4.09万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
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/10656221
关键词：
Adenosine Monophosphate Affect Aging Alleles Animals Attenuated Biochemical Biological Assay Buffers CAG repeat 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 Molecular Molecular Chaperones Mutagenesis Neurodegenerative Disorders Neurons Nucleotides 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 Techniques Testing Therapeutic Therapeutic Intervention Tissues Toxic effect Type 1 Spinocerebellar Ataxia Work arm biological adaptation to stress derepression disease-causing mutation follow-up 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

项目摘要

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.

项目摘要亨廷顿氏病（HD）和脊椎脑性共济失调3型（SCA3）是遗传的衰老相关疾病对患者和家庭看护人的普遍性造成了毁灭性影响人口。这些疾病属于由聚谷氨酰胺（PolyQ）膨胀疾病的家族突变导致三核苷酸（CAGN）在不同基因中重复的病理扩张。增加 CAG重复长度产生含有异常扩张的Polyq曲线的蛋白质，干扰正常蛋白质功能并促进错误折叠。这些疾病的毒性被认为部分是由于形成而产生的包括异常构象突变蛋白的病理纳入体，在许多与衰老相关的神经退行性疾病。尽管大力努力破译机制但是，在PolyQ疾病中的潜在毒性，但是，在确定目标的目标方面几乎没有取得进展治疗干预。最近，翻译后修饰（PTM），两极化已成为HSP70的新型调节剂家族伴侣，细胞蛋白质质量控制机制的关键组成部分，可缓冲蛋白质错误折叠的压力。蛋白质安培化是由FIC型安培酶，人类FICD及其直系同源物进行的 fic-1，分别在秀丽隐杆线虫中。我们实验室中的工作已经确定，FIC-1介导的聚氨酯直接改变 PolyQ聚集动力学和毒性。此外，我的初步数据列出了 FIC-1缺乏症足以挽救在表达聚集polyqs的秀丽隐杆线虫的生存期间以polyq长度依赖性方式发展。综上所述，这些发现表明损失 1/FICD介导的双甲基化Bolsters蛋白技术网络能力减轻PolyQ蛋白诱导的毒性聚合。该项目将利用跨学科方法来产生FICD/FIC-1-的整体表征 PolyQ疾病中介导的聚氨酯。为此，我将利用秀丽隐杆线虫的强大遗传学揭露面对致病PolyQ聚集的新途径，可促进生存（AIM 1）。在同时，我会源自HD和SCA3患者干细胞的神经元中的员工功能测定，以介绍FICD活性在这些疾病模型中影响PolyQ聚集和毒性（AIM 2）。这些研究的结果将进步我们对两极化如何调节PolyQ疾病中蛋白质的知识。我研究的最终目标是利用这些发现，以开发可翻译的治疗方法，以用于衰老相关疾病。