An Alternative Isoform of RRM2B as a Genetic Modifier in Huntington's Disease

RRM2B 的替代亚型作为亨廷顿病的遗传修饰剂

基本信息

批准号：
10621765
负责人：
Ihn Sik Seong
金额：
$ 39.48万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621765
关键词：
Affect Age of Onset Alleles Amino Acids Antisense RNA Binding Biochemical Biological Biological Assay Biological Models Biology CAG repeat CRISPR/Cas technology Cell Differentiation process Cell Survival Cell model Cells Cellular Stress Characteristics Chemicals Chromosomes Clustered Regularly Interspaced Short Palindromic Repeats Complex DNA Maintenance Data Diphosphates Disease Drug Design Engineering Exons Foundations Future Gene Expression Gene Transfer Genes Genetic Genetic study Goals Haplotypes Homeostasis Human Huntington Disease Huntington gene Inherited Investigation Knock-out Knockout Mice Label Length Maintenance Mediating Methods Mitochondria Mitochondrial DNA Mitochondrial DNA depletion syndromes Modification Molecular Mutation Neurodegenerative Disorders Neuronal Differentiation Neurons Onset of illness Outcome Oxidoreductase Pathogenesis Pathogenicity Pathologic Pathway interactions Patients Persons Phenotype Play Ploidies Primates Protein Isoforms Proteins Proteomics RRM1 gene Regulation Resources Rest Ribonucleosides Ribonucleotide Reductase Role Sample Size Series Signal Transduction Source Stress TP53 gene Techniques Testing Therapeutic Therapeutic Effect Therapeutic Intervention Time Trinucleotide Repeat Expansion Untranslated RNA Variant Work biological adaptation to stress design disease phenotype exon skipping expectation experimental study follow-up gene therapy genetic analysis genome wide association study induced pluripotent stem cell innovation insight knock-down mitochondrial dysfunction motor symptom nerve stem cell nervous system disorder neuron loss new technology novel novel therapeutics polyglutamine premature prevent protein complex recognins response small hairpin RNA spatiotemporal stressor therapeutic candidate therapeutic development tool transcriptome ubiquitin-protein ligase

项目摘要

The pathogenic mechanism of Huntington’s disease (HD), a fatal, dominantly inherited neurodegenerative disorder caused by trinucleotide repeat expansion in the huntingtin (HTT) gene, is poorly understood, stymieing therapeutic development. Our overarching goal is to better understand HD pathogenesis and identify targetable pathways for therapeutic intervention by studying genetic modifiers of HD, initially identified by the GeM-HD consortium. The objective of this proposal is to evaluate one HD modifier, ribonucleoside-diphosphate reductase subunit M2 B (RRM2B), by delineating its isoform-specific biological activity on mitochondrial regulation and characterizing its effect on HD pathogenesis. Our hypothesis is that the two SNPs in chr8 GWAS modifier haplotype, rs1037699 and rs5893603, shown to lower RRM2B isoform 2 levels, play a critical role in isoform 2- specific complex(es), altering overall RRM2B activity and HD pathogenesis. Upon completion, this work will validate RRM2B isoform 2 as a modulator of HD pathogenesis and serve as a foundation for novel therapeutic disease-altering strategies as well as providing insight into the fundamental mechanism of HD by: 1) Characterizing isoform-specific RRM2B activity, focusing on its role in mitochondrial regulation, on subcellular localization, stress-response assays, and knockdown experiments in LCLs and human neuronal progenitor cells (hNPCs) and neurons differentiated from our new HTT isogenic series of iPSCs with different CAG sizes (17, 40, 51 and 61); 2) Identifying key cellular protein complexes containing RRM2B isoform 2 in a stress and time dependent manner using APEX-mediated quantitative proteomics; 3) Evaluating potential therapeutic effects by augmenting RRM2B isoform 2 levels in HD patient cells using novel technologies, such as non-coding antisense RNA (SIENUP). Novel techniques and resources include sensitive allele-specific SRM-MS for targeted quantification, recently optimized gene therapy tools, and innovative HD cell model systems, designed to precisely delineate CAG signatures relevant for HD. This proposal will unveil the interface between RRM2B biology and HD pathogenesis, establishing RRM2B as a targetable pathway and potentially revealing other players in the pathogenic cascade. Our expectation is that strategies to upregulate RRM2B isoform 2 level will have a positive effect on HD phenotypes, providing a key candidate for therapeutics. Overall, the positive outcomes of this work will enable us to better understand the effect of genetic modifiers on HD and validate immediate targets for rational drug design, providing additional avenues for future therapeutic strategies.

亨廷顿病（HD）的致病机制，一种致命的显性遗传性神经退行性疾病亨廷顿蛋白 (HTT) 基因中三核苷酸重复扩增引起的疾病目前尚不清楚，阻碍了我们的首要目标是更好地了解 HD 发病机制并确定有针对性的治疗方法。通过研究 HD 基因修饰物进行干预的治疗途径，最初由 GeM-HD 确定该提案的目的是评估一种 HD 修饰剂，核糖核苷二磷酸还原酶。亚基 M2 B (RRM2B)，通过描绘其对线粒体调节的异构体特异性生物活性和表征其对 HD 发病机制的影响我们的假设是 chr8 GWAS 修饰符中的两个 SNP。单倍型 rs1037699 和 rs5893603 显示可降低 RRM2B 同工型 2 水平，在同工型 2- 中发挥关键作用特定复合物，改变总体 RRM2B 活性和 HD 发病机制。完成后，这项工作将。验证 RRM2B 同工型 2 作为 HD 发病机制的调节剂并作为新型治疗的基础改变疾病的策略以及通过以下方式深入了解 HD 的基本机制：1) 表征异构体特异性 RRM2B 活性，重点关注其在线粒体调节和亚细胞中的作用 LCL 和人类神经祖细胞的定位、应激反应测定和敲低实验 (hNPC) 和从我们新的 HTT 同基因系列 iPSC 中分化出来的神经元，这些 iPSC 具有不同的 CAG 大小（17、40、 51 和 61)；2) 在压力和时间下鉴定含有 RRM2B 同工型 2 的关键细胞蛋白复合物使用 APEX 介导的定量蛋白质组学的依赖方式；3) 评估潜在的治疗效果使用非编码反义等新技术增强 HD 患者细胞中的 RRM2B 同工型 2 水平 RNA (SIENUP) 新技术和资源包括用于靶向的敏感等位基因特异性 SRM-MS。定量、最近优化的基因治疗工具和创新的 HD 细胞模型系统，旨在精确描述与 HD 相关的 CAG 签名该提案将揭示 RRM2B 之间的接口。生物学和 HD 发病机制，将 RRM2B 建立为可靶向途径，并可能揭示其他我们期望上调 RRM2B 同工型 2 水平的策略将会发挥作用。对 HD 表型有积极影响，为治疗提供了一个关键的候选者。这项工作的成果将使我们能够更好地了解基因修饰剂对 HD 的影响并验证合理药物设计的直接目标，为未来的治疗策略提供额外的途径。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Genetic modifiers of Huntington disease differentially influence motor and cognitive domains.

亨廷顿病的基因修饰对运动和认知领域有不同的影响。

DOI：
发表时间：
2022-05-05
期刊：
影响因子：
9.8
作者：
Lee, Jong;Huang, Yuan;Orth, Michael;Gillis, Tammy;Siciliano, Jacqueline;Hong, Eunpyo;Mysore, Jayalakshmi Srinidhi;Lucente, Diane;Wheeler, Vanessa C;Seong, Ihn Sik;McLean, Zachariah L;Mills, James A;McAllister, Branduff;Lobanov, Sergey V
通讯作者：
Lobanov, Sergey V