Nucleocentric molecular characterization of C9orf72 NRE-linked neurodegeneration

C9orf72 NRE 相关神经变性的核中心分子表征

• 批准号: 9444083
• 负责人: Aaron Raymond Haeusler
• 金额: $ 24.9万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9444083
• 关键词: Address; Adopted; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Ataxia; Base Sequence; Binding; Binding Proteins; Biochemical; C9ORF72; Cells; Chromosomal Rearrangement; Chromosome Fragile Sites; Chromosomes, Human, Pair 9; Chronic; Clinical; Complex; Coupling; DNA; DNA Damage; DNA Repeat Expansion; DNA Sequence; DNA Sequence Alteration; DNA-Protein Interaction; Data; Defect; Disease; Doctor of Philosophy; Drosophila genus; Fragile X Syndrome; Frontotemporal Dementia; G-Quartets; Genome Stability; Genomic Instability; Goals; Hereditary Disease; Huntington Disease; Hybrids; In Vitro; Lead; Libraries; Ligand Binding; Ligands; Link; Mentors; Methods; Modeling; Molecular; Molecular Analysis; Molecular Conformation; Motor Neurons; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Nucleoproteins; Nucleotides; Patients; Phase; Phenocopy; Postdoctoral Fellow; Process; Proteins; RNA; Reading Frames; Research; Ribonucleoproteins; Series; Site; Stress; Structure; Techniques; Testing; Therapeutic; Tissues; Toxic effect; Transgenes; Transgenic Organisms; Work; age related; biophysical properties; disease phenotype; experimental study; gain of function; in vivo; in vivo Model; infancy; meetings; nervous system disorder; novel; nucleic acid structure; prevent; protein complex; public health relevance; screening; stem; therapeutic target

 DESCRIPTION (provided by applicant): A genetic mutation that leads to a nucleotide repeat expansion (NRE) of repetitive DNA on chromosome 9 in protein reading frame 72 (C9orf72) has been associated with a number of age-related neurodegenerative diseases: Lou Gehrig's disease (ALS), frontal temporal dementia (FTD), Alzheimer's disease, and a phenocopy of Huntington's disease. Recent advances in our molecular understanding of the C9orf72 NRE mutation has demonstrated this NRE also shares many cellular defects found in other NRE-linked neurological diseases, such as Fragile X, many ataxias, and Huntington's disease. Therefore, the research proposed here will examine the molecular defects caused by NREs using the C9orf72 NRE paradigm. The research will investigate three important aspects to advance the understanding of NRE-linked defects. First, proteins that bind to the non-canonical structures formed by the DNA and RNA of the C9orf72-NRE will be examined for their mechanism(s) of recognition, and the cellular defects that result from the nucleotide*protein complex will be determined in patients cells and tissues. Second, the non-canonical nucleotide structures formed by the C9orf72 NRE can endanger genome stability. Therefore, the relationship between nucleotide structures and DNA damage for the C9orf72 NRE will be addressed using in vitro biochemical experiments, and examined in patient cells and tissues. The final work proposed here will identify ligands that bind to non-canonical nucleotide structures formed by the C9orf72 NRE to prevent the downstream NRE-linked cellular defects. This will be performed using a thorough biophysical characterization of nucleotide*ligand complexes and nucleotide*protein complexes in the presence of ligands. Promising ligand candidates will then be examined for their efficacy in patient induce-pluripotent stem motor neuron cells and in Drosophila expressing a transgenic C9orf72 NRE. In summary, the proposed work will advance our molecular understanding of the chronic molecular stresses caused by the C9orf72 NRE that lead to the age-related neurodegenerative defects, and it will identify potential disease modifying ligands.

 描述(申请人提供)：一种导致蛋白质阅读框架72(C9orf72)中9号染色体上重复DNA核苷酸重复扩展(NRE)的基因突变与许多与年龄相关的神经退行性疾病有关：卢·格里克病(ALS)、额叶颞叶痴呆(FTD)、阿尔茨海默病和亨廷顿病的表型。我们对C9orf72 NRE突变的分子理解的最新进展表明，该NRE也存在许多与NRE相关的神经系统疾病的细胞缺陷，如脆性X、许多共济失调和亨廷顿病。因此，这里提出的研究将使用C9orf72 NRE范式来检查NRES引起的分子缺陷。这项研究将从三个重要方面来推进对NRE相关缺陷的理解。首先，将检查与C9orf72-NRE的脱氧核糖核酸和核糖核酸形成的非规范结构结合的蛋白质的识别机制(S)，并将在患者细胞和组织中确定由核苷酸*蛋白质复合体导致的细胞缺陷。其次，C9orf72 NRE形成的非规范核苷酸结构可能危及基因组的稳定性。因此，C9orf72 NRE的核苷酸结构和DNA损伤之间的关系将通过体外生化实验来解决，并在患者细胞和组织中进行检测。本文提出的最后一项工作将确定与C9orf72 NRE形成的非规范核苷酸结构结合的配体，以防止下游NRE连接的细胞缺陷。这将通过在存在配体的情况下对核苷酸*配体复合体和核苷酸*蛋白质复合体进行彻底的生物物理表征来实现。有希望的候选配体将在患者诱导的多能干细胞运动神经元细胞和表达转基因C9orf72 NRE的果蝇中进行有效性测试。综上所述，这项拟议的工作将促进我们对C9orf72 NRE引起的慢性分子应激反应的分子理解，并将识别潜在的疾病修饰配体。