Somatic mtDNA mutations in brain Aging: a single-cell approach

大脑衰老中的体细胞 mtDNA 突变：单细胞方法

基本信息

批准号：
7806584
负责人：
Konstantin Khrapko
金额：
$ 30.43万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-09-30 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7806584
关键词：
Affect Age Aging Area Brain Brain Part Cells Cerebellum Cerebral cortex Clonal Expansion Collaborations Collection Consensus Cytogenetics DNA DNA Sequence Data Defect Deterioration Development Disease Employee Strikes Epidemiology Fluorescent in Situ Hybridization Framingham Heart Study Funding Genetic Genomics Goals Health Heart Hippocampus (Brain)Human Individual Inherited Institutes Interdisciplinary Study Longevity Measurement Medicine Memory Loss Mental Depression Methodology Methods Mitochondria Mitochondrial DNA Mitochondrial Diseases Monitor Movement Mus Mutation Mutation Detection Nature Nerve Degeneration Neurons Oxidative Stress Parkinson Disease Parkinsonian Disorders Patients Phenotype Pigments Point Mutation Population Power Plants Premature aging syndrome Prevalence Procedures Process Purkinje Cells Research Research Personnel Respiratory Chain Role Sampling Specialist Structure of choroid plexus Substantia nigra structure Symptoms Techniques Technology Testing Tissues Variant Work age related aged aging brain cell type color detection cost empowered interest medical schools mitochondrial DNA mutation motor impairment mutant new technology novel programs putamen respiratory single molecule success

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of the proposed research is to study the mechanisms responsible for age-related degenerative processes. The understanding of these mechanisms may help to find ways to slow down the corresponding processes thus moving the onset of deterioration outside the normal human lifespan. Accumulation of mtDNA mutations has been long hypothesized as a probable cause of the various symptoms of aging, however, proof of such involvement was lacking. There is no consensus regarding which tissues and/or cell types if any are most likely to be affected. Our recent research has demonstrated unprecedented accumulation of mtDNA deletions in the substantia nigra of the aged human brain, which results in respiratory chain defects in pigmented neurons. This demonstration is important conceptually as an example the most significant involvement of somatic mtDNA mutations in an age-related degenerative process, but it also raises questions regarding the health consequences of this process, and whether it is limited to s. nigra. The specific aims therefore are: (1) To test the hypothesis that clonal expansions of mtDNA mutations, either deletions or point mutations, cause functional defects in specific cell types of the brain, and/or cause degeneration or predispose cells to degeneration. (2) To test the hypothesis that accumulation of mtDNA deletions in pigmented neurons of substantia nigra contributes to the development of Mild Parkinsonian Signs in seniors. We will compare the prevalence of cells with mtDNA deletions in a collection of well- characterized brains with respect to the presence of parkinsonian signs. (3) To develop new methodologies for mtDNA mutation quantification in individual cells. The methods include mtDNA Fluorescent In Situ Hybridization (FISH) for quantification of mtDNA deletions, and a novel massive parallel 454 DNA sequencing approach for studying mtDNA point mutations. In short, we have discovered a novel, common, age-related degenerative process in the brain involving the DNA of mitochondria, the power plants of the cell. This process affects an area involved in Parkinson's disease. We are therefore exploring if the process is responsible for movement problems widespread among the senior population. We are also testing whether similar processes affect other parts of the aging brain.

描述（由申请人提供）：拟议研究的长期目标是研究负责年龄相关的退行过程的机制。对这些机制的理解可能有助于找到减慢相应过程的方法，从而将恶化的开始在正常的人类寿命之外移动。长期以来，长期以来，MTDNA突变的积累是衰老症状的可能原因，但是缺乏这种参与的证据。关于哪些组织和/或细胞类型（如果有可能受到的影响）尚无共识。我们最近的研究表明，在老年人大脑的底虫中，mtDNA缺失的前所未有，这导致有色素神经元的呼吸链缺陷。从概念上讲，这一演示是一个重要的例子，是体细胞mtDNA突变在与年龄相关的退化过程中最重要的参与，但它也引发了有关此过程的健康后果以及是否仅限于s的疑问。尼格拉。因此，具体目的是：（1）检验以下假设：mtDNA突变的克隆膨胀（缺失或点突变）在大脑的特定细胞类型中引起功能缺陷，以及/或导致退化或倾向细胞变性。（2）检验以下假设：黑质的色素神经元中mtDNA缺失的积累有助于发展老年人中温和的帕金森氏症体征。我们将在帕金森氏症迹象的存在方面比较细胞的流行与mtDNA缺失。（3）开发单个细胞中mtDNA突变定量的新方法。这些方法包括用于定量mtDNA缺失的原位原位杂交（FISH），以及用于研究mtDNA点突变的新型大规模平行454 DNA测序方法。简而言之，我们在大脑中发现了一个新颖的，常见的，年龄相关的退化过程，涉及线粒体的DNA，即细胞的发电厂。这个过程影响帕金森氏病涉及的领域。因此，我们正在探索该过程是否负责运动问题在老年人中普遍存在。我们还在测试类似的过程是否影响大脑衰老的其他部位。