Investigation of ALS caused by mutant CHCHD10

CHCHD10 突变体引起的 ALS 的研究

基本信息

批准号：
9983200
负责人：
TEEPU SIDDIQUE
金额：
$ 50.73万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9983200
关键词：
ALS patients Affect Amyotrophic Lateral Sclerosis Animal Model Axon Axonal Transport Behavior Behavioral Binding Binding Proteins Biochemical Process Biological Models Biology Brain Buffers CRISPR/Cas technology Calcium Cell physiology Cells Cellular biology Data Defect Degenerative Disorder Disease Electron Microscopy Engineering Etiology Familial Amyotrophic Lateral Sclerosis Family Genes Genetic Genetic Engineering Goals Human Impairment Inherited Investigation Knock-in Laboratories Laboratory Personnel Link Membrane Potentials Mitochondria Mitochondrial Swelling Modeling Molecular Target Morphology Motor Neurons Mus Mutation Nature Neurodegenerative Disorders Neurons Nuclear Outcome Oxidation-Reduction Oxidative Phosphorylation Pathogenicity Pathologic Pathology Patients Periodicity Phenotype Physiological Positioning Attribute Prevention Process Proteins Reagent Research Resources Rodent Model Role Spinal Cord Subgroup Surveys Swelling System Techniques Testing Tissues Transgenic Mice base cell type disease mechanisms study disease-causing mutation experimental study gain of function human disease in vivo induced pluripotent stem cell insight knock-down mitochondrial dysfunction motor deficit motor impairment mouse model mutant nervous system disorder novel overexpression response skills trait

项目摘要

Some of the most robust advances in the understanding of amyotrophic lateral sclerosis (ALS), a multi- etiologic and fatal disorder of the nervous system, have come from the identification of the genetic basis of a subgroup of ALS cases. In this subgroup of ALS (familial ALS or FALS), the disease is inherited as a familial trait. Identification of a single mechanism of disease across all of ALS has been elusive, but notable amongst proposed common mechanisms is mitochondrial dysfunction. However, till recently, no direct robust etiological link between mitochondrial genes, nuclear or mitochondrial, had been found. Recently, others and we found that mutations in a gene called CHCHD10 causes ALS and additional phenotypes. Some other ALS genes, including VCP, HNRNPA1, SQSTM1 and OPTN are also associated with other phenotypes besides ALS. CHCHD10 and its protein product are not well studied. To investigate how a mutation in CHCHD10 may cause ALS, we have collected or developed promising reagents and assembled collaborators and laboratory personnel with specialized skills in genetic engineering, rodent models, cell biology, CRISPR/Cas9 gene editing, mitochondrial function, redox response, induced pluripotent stem cell (iPSC) derived motor neurons and primary motor neurons to carry out the proposed experiments in my laboratory and the laboratories of collaborators. We will investigate the pathology and behavior of a new transgenic mouse model we have engineered to overexpress the mutant (R15L) human CHCHD10 gene. Preliminary study shows axonal pathology in the spinal cord and brain of this mouse with periodic beaded axonal swellings harboring mitochondria. Additional more precise models will also be developed. To determine what aspect of mitochondrial function or morphology is affected by mutant CHCHD10 we will screen mitochondrial function of energetics, redox studies, calcium buffering and electron microscopy analysis. We will identify the binding protein partner(s) of CHCHD10 protein to understand the role of CHCHD10 in the mitochondrial biology. On completion, our study would provide a clearer understanding of the central defect(s) in this form of ALS and potentially a more granular understanding of mitochondrial defect generalizable across ALS. The study will also allow the identification of potential molecular targets for rational therapy and/or prevention of ALS.

在理解肌萎缩侧索硬化症（ALS）方面取得的一些最强有力的进展，肌萎缩侧索硬化症是一种多发性硬化症。神经系统的病因学和致命性疾病，来自对遗传基础的鉴定 ALS 病例亚组。在 ALS（家族性 ALS 或 FALS）的这个亚组中，该疾病是家族性遗传的特征。识别所有 ALS 的单一疾病机制一直难以捉摸，但值得注意的是提出的共同机制是线粒体功能障碍。然而，直到最近，还没有直接可靠的病因学线粒体基因（核基因或线粒体基因）之间的联系已经被发现。最近，其他人和我们发现 CHCHD10 基因突变会导致 ALS 和其他表型。其他一些 ALS 基因，包括VCP、HNRNPA1、SQSTM1和OPTN也与ALS以外的其他表型相关。 CHCHD10 及其蛋白质产物尚未得到充分研究。研究 CHCHD10 突变如何导致 ALS，我们收集或开发了有前途的试剂并组建了合作者和实验室具有基因工程、啮齿动物模型、细胞生物学、CRISPR/Cas9基因专业技能的人员编辑、线粒体功能、氧化还原反应、诱导多能干细胞 (iPSC) 衍生的运动神经元和初级运动神经元在我的实验室和实验室进行拟议的实验合作者。我们将研究我们设计的新转基因小鼠模型的病理学和行为过度表达突变 (R15L) 人 CHCHD10 基因。初步研究显示轴突病理学这只小鼠的脊髓和大脑具有周期性的串珠状轴突肿胀，其中含有线粒体。额外的还将开发更精确的模型。确定线粒体功能的哪个方面或形态受到突变体 CHCHD10 的影响，我们将筛选能量学、氧化还原研究的线粒体功能，钙缓冲和电子显微镜分析。我们将鉴定 CHCHD10 的结合蛋白伴侣蛋白质以了解 CHCHD10 在线粒体生物学中的作用。完成后，我们的研究将提供对这种 ALS 形式的中心缺陷的更清晰的理解，并可能提供更细化的信息对线粒体缺陷的理解可推广到 ALS。该研究还将确定合理治疗和/或预防 ALS 的潜在分子靶点。