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（由申请人提供）：导致 9 号染色体上蛋白质阅读框 72 (C9orf72) 重复 DNA 核苷酸重复扩增 (NRE) 的基因突变与许多与年龄相关的神经退行性疾病有关：卢伽雷氏病(ALS)、额颞叶痴呆 (FTD)、阿尔茨海默病和亨廷顿舞蹈病表型对 C9orf72 的分子理解的最新进展。 NRE 突变已证明该 NRE 也具有其他与 NRE 相关的神经系统疾病中发现的许多细胞缺陷，例如脆性 X 病、许多共济失调和亨廷顿舞蹈病。因此，本文提出的研究将使用 C9orf72 NRE 检查由 NRE 引起的分子缺陷。该研究将研究三个重要方面，以促进对 NRE 相关缺陷的理解。首先，与 DNA 和 RNA 形成的非规范结构结合的蛋白质。将检查 C9orf72-NRE 的识别机制，并确定患者细胞和组织中由核苷酸*蛋白质复合物引起的细胞缺陷。其次，可以确定由 C9orf72 NRE 形成的非规范核苷酸结构。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查。本文提出的最终工作将确定。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。这将在配体存在的情况下使用核苷酸*配体复合物和核苷酸*蛋白质复合物的彻底生物物理表征来进行。然后将检查配体候选物在患者诱导多能干运动神经元细胞和表达转基因 C9orf72 NRE 的果蝇中的功效。这项工作将增进我们对 C9orf72 NRE 引起的慢性分子应激（导致与年龄相关的神经退行性缺陷）的分子理解，并将识别潜在的疾病修饰配体。