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.

在理解肌萎缩性侧索硬化症（ALS）方面，一些最强大的进步，神经系统的病因和致命疾病来自于遗传基础的鉴定 ALS案例的亚组。在这个ALS的亚组（家族性ALS或FALS）中，该疾病被遗传为一种家族性特征。在所有ALS中识别疾病的单一机制一直难以捉摸，但在提出的共同机制是线粒体功能障碍。但是，直到最近，还没有直接鲁棒的病因线粒体基因（核或线粒体）之间的联系已被发现。最近，其他人发现在称为CHCHD10的基因中的突变会引起ALS和其他表型。其他一些ALS基因，包括VCP，HNRNPA1，SQSTM1和OPTN在内的其他表型也与ALS之外的其他表型相关联。 CHCHD10及其蛋白质产物尚未得到很好的研究。研究CHCHD10中的突变如何导致 ALS，我们已经收集或开发了有前途的试剂以及组装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基因的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原发性运动神经元，以在我的实验室和实验室中进行拟议的实验合作者。我们将研究已设计为新的转基因鼠标模型的病理和行为过表达突变体（R15L）人CHCHD10基因。初步研究显示了轴突病理这款小鼠的脊髓和大脑带有带有线粒体的周期性串珠轴突肿胀。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突变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。我们将确定CHCHD10的结合蛋白伴侣蛋白质了解CHCHD10在线粒体生物学中的作用。完成后，我们的研究将以这种ALS形式更清楚地了解中央缺陷，并可能更细粒度了解ALS可推广的线粒体缺陷。该研究还将允许识别有理治疗和/或预防ALS的潜在分子靶标。