Characterization of a Novel Ataxia Phenotype

新型共济失调表型的表征

• 批准号: 10400578
• 负责人: Alexander Telenson
• 金额: $ 4.23万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10400578
• 关键词: Affect; Alleles; Animal Model; Animal Sources; Ataxia; Backcrossings; Bioinformatics; Biological Process; Brain Stem; Breeding; Candidate Disease Gene; Cells; Cerebellum; Cessation of life; Chromosome Mapping; Computer Analysis; Critical Pathways; Data; Defect; Development; Disease; Embryonic Development; Ensure; Equilibrium; Essential Genes; Fluorescent in Situ Hybridization; Gene Expression; Generations; Genes; Genetic; Genetic Complementation Test; Genetic Variation; Genome; Genotype; Goals; Growth; Human; Inherited; Light; Longevity; Missense Mutation; Modeling; Molecular; Mus; Mutant Strains Mice; Mutation; Neuraxis; Neurologic; Nonsense Mutation; Nutritional; Pathology; Pattern; Phenotype; Play; Principal Investigator; Probability; Process; Role; Severities; Slice; Spinal Ganglia; Stains; Thinness; Time; Tissues; Training; Untranslated RNA; Validation; Variant; causal variant; cell type; design; differential expression; experimental study; genetic approach; genetic variant; genome analysis; genome sequencing; human disease; insight; mouse model; mutant; neurodevelopment; neuromuscular; novel; postnatal; postnatal development; postnatal period; segregation; skills training; success; therapeutic development; transcriptome sequencing; transcriptomics; whole genome

PROJECT SUMMARY/ABSTRACT Defining genetic variation responsible for neuromuscular phenotypes has provide insight about basic neurological mechanisms as well as insight on human disease mechanisms. Mouse models have also been used to understand genetic mechanisms that cause relevant neurological conditions. These discoveries were made through the classic genetic approach of characterizing a phenotype and then defining the genotype that accounts for that defect. This approach has been instrumental for a host of discoveries resulting in a more thorough understanding of underlying mechanisms of disease. The project put forth under this proposal follows a similar approach. We seek to identify and characterize a potentially novel variant responsible for a severe, “novel ataxia phenotype” (nap) discovered in our lab with the ultimate goal of shedding light on novel mechanisms underlying ataxia. This proposal outlines experiments to characterize the affected tissue type and evaluate whole genome sequencing (WGS) and RNA sequencing to isolate potentially causal genetic variants to experimentally validate. This approach ensures that progress is not wholly dependent on a specific result, rather several experiments can be done in concert that will increase the probability of success. Our approach of utilizing WGS will providing the genetic landscape thereby elucidating potential noncoding variants that may elicit the phenotype in the nap mice. After determining which tissue type is affected, we will perform RNA sequencing to elucidate transcriptomic differences in the nap mice to provide potential mechanisms underlying the phenotype. Successfully identifying the variant through computationally analysis and experimental validation will allow for further experiments to better understand the mechanism of action of the variant, as outlined in the aims. Given the presentation of the ataxia phenotype, we suspect the primary tissue affected will be the cerebellum and/or brainstem and through breeding, we have established that this variant is inherited in an autosomal recessive pattern. Through preliminary genome analysis, we have identified several novel candidates and excluded several known genes responsible for ataxia phenotypes. The results of this project are expected to provide further insight into the molecular mechanisms that underlie ataxia and will potentially serve as a model for therapeutic development to ameliorate ataxia symptomology. This project was designed to expand the skills and training of the principal investigator with the goal of comprehensive training in bioinformatics and characterization of the mouse model.

项目概要/摘要 定义导致神经肌肉表型的遗传变异提供了关于基本原理的见解 神经机制以及对人类疾病机制的见解。鼠标模型也已 用于了解导致相关神经系统疾病的遗传机制。这些发现是 通过经典的遗传方法来表征表型，然后定义基因型 解释了这个缺陷。这种方法对于许多发现发挥了重要作用，从而产生了更多 深入了解疾病的潜在机制。根据本提案提出的项目如下 类似的方法。我们试图识别和表征一种潜在的新变体，该变体可导致严重的、 我们实验室发现了“新的共济失调表型”（nap），最终目标是揭示新的共济失调表型 共济失调的潜在机制。该提案概述了表征受影响组织类型和 评估全基因组测序 (WGS) 和 RNA 测序以分离潜在的致病遗传变异 来进行实验验证。这种方法确保进展并不完全依赖于特定结果， 相反，可以同时进行多个实验，这将增加成功的可能性。我们的方法 利用 WGS 将提供遗传景观，从而阐明可能的潜在非编码变异 引发午睡小鼠的表型。确定受影响的组织类型后，我们将进行 RNA 测序以阐明小睡小鼠的转录组差异，以提供潜在的机制 表型。通过计算分析和实验成功识别变体 验证将允许进一​​步的实验，以更好地了解该变体的作用机制，因为 目标中概述。鉴于共济失调表型的表现，我们怀疑受影响的主要组织 将是小脑和/或脑干，通过育种，我们已经确定这种变异是遗传的 呈常染色体隐性遗传模式。通过初步的基因组分析，我们发现了几种新的 候选者并排除了几个已知的导致共济失调表型的基因。该项目的成果 预计将进一步深入了解共济失调的分子机制，并有可能 作为改善共济失调症状的治疗开发模型。这个项目是设计的 扩大主要研究者的技能和培训，目标是全面培训 小鼠模型的生物信息学和表征。