Nucleocentric molecular characterization of C9orf72 NRE-linked neurodegeneration

C9orf72 NRE 相关神经变性的核中心分子表征

基本信息

批准号：
8869973
负责人：
Aaron Raymond Haeusler
金额：
$ 9.42万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8869973
关键词：
Address Adopted Alzheimer&apos s Disease Amyotrophic Lateral Sclerosis Ataxia Base Sequence Binding Biochemical Cells Chromosomal Rearrangement Chromosome Fragile Sites Chromosomes, Human, Pair 9 Chronic 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 Protein Binding 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 gain of function in vivo in vivo Model infancy meetings nervous system disorder novel nucleic acid structure prevent protein complex public health relevance research study screening stem therapeutic target

项目摘要

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（由适用提供）：一种遗传突变，导致蛋白质阅读框架72（C9ORF72）在9染色体上的核重复膨胀（NRE）与年龄相关的神经退行性疾病有关疾病。我们对C9ORF72 NRE突变的分子理解的最新进展表明，这种NRE在其他NRE连接的神经系统疾病中也有许多细胞缺陷，例如脆弱的X，许多共济失调和亨廷顿病。因此，这里提出的研究将检查使用C9orf72 NRE范式引起的NRES引起的分子缺陷。该研究将研究三个重要方面，以提高对NRE连接缺陷的理解。首先，将检查由C9ORF72-NRE的DNA和RNA形成的非典型结构的蛋白质的识别机制，并在患者细胞和组织中确定由核苷酸*蛋白质复合物引起的细胞缺陷。其次，由C9ORF72 NRE形成的非传统核苷酸结构可能危害基因组稳定性。因此，C9ORF72的核苷酸结构与DNA损伤之间的关系和此处提出的最终工作将确定与C9orf72 NRE形成的非典型核丁基结构结合的配体，以防止下游NRE连接的细胞缺陷。这将使用核丁物*配体复合物和核丁基*蛋白质复合物的彻底生物物理表征进行。然后，将检查有希望的配体候选物在患者中诱导型茎运动神经元细胞和表达转基因C9orf72 NRE的果蝇的有效性。总而言之，拟议的工作将提高我们对由C9ORF72 NRE引起的慢性分子应激的分子理解，该慢性分子应力导致与年龄相关的神经退行性缺陷，并将鉴定潜在的疾病修饰配体。