NICOTINAMIDE MONONUCLEOTIDE ADENYLYLTRANSFERASES IN NEONATAL HYPOXIA-ISCHEMIA

烟酰胺单核苷酸腺苷酸转移酶在新生儿缺氧缺血中的作用

基本信息

批准号：
8692037
负责人：
Rafael Galindo
金额：
$ 17.28万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8692037
关键词：
Accounting Acute Affect Animal Model Animals Axon Biochemical Biological Process Brain Brain Hypoxia-Ischemia Brain Injuries Brain region Buffers Calcium Cause of Death Cell Death Cells Cerebral Palsy Cessation of life Chronic Clinical Development Disease Enzymes Epilepsy Family Fluorescent Dyes Functional disorder Gene Expression Genes Genetic Techniques Glutamate Receptor Glutamates Goals High Prevalence Homologous Gene Human Hypoxia Injury Intellectual functioning disability Intervention K-Series Research Career Programs Knock-out Knowledge Laboratories Lead Leber&apos s amaurosis Light Mediating Membrane Potentials Mentors Mentorship Methods Mitochondria Modeling Molecular Genetics Mutation Neonatal Neonatal Brain Injury Nerve Degeneration Neuraxis Neurobiology Neurodegenerative Disorders Neurologic Neurons Newborn Infant Nicotinamide adenine dinucleotide Nicotinamide-Nucleotide Adenylyltransferase Pathway interactions Pattern Perinatal Peripheral Peripheral Nerves Physicians Physiology Play Prevention Prevention strategy Process Property Proteins Receptor Activation Regulation Retina Role Scientist System Techniques Testing Time Transduction Gene Transgenic Animals Transgenic Model Viral Western Blotting base effective therapy excitotoxicity in vitro Model in vivo injured knock-down mitochondrial membrane mouse model neonatal hypoxic-ischemic brain injury neonate nerve injury nervous system disorder neurobiological mechanism neuron loss neuronal survival neuroprotection new therapeutic target novel therapeutics overexpression public health relevance research study vector

项目摘要

DESCRIPTION (provided by applicant): The goal of this Mentored Career Development Award is to facilitate Dr. Rafael Galindo's transition to independence as a physician-scientist studying the neurobiological mechanisms of neuronal death in injured developing neurons. The proposed experiments will be conducted under the mentorship of Dr. David Holtzman, and they will examine the potential neuroprotective role of a family of three NAD+ metabolizing enzymes, known as nicotinamide mononucleotide adenylyltransferases (NMNATs), in neonatal brain hypoxia- ischemia (H-I). This form of injury is the most common cause of death in the newborn period and accounts for a significant portion of the chronic neurological conditions attributed to brain injury in the perinatal period. Despite its high prevalence, there are only limited effective therapies available for newborns exposed to H-I. This lack of effective clinical intervention is attributed, in part, to our incomplete understanding of the neurobiological mechanisms and molecules that regulate neuronal death pathways in the healthy and injured developing brain. NMNATs are molecules that have been shown to have strong neuroprotective properties and are likely importantly involved in the neurodegenerative mechanisms of various central and peripheral neurological diseases. Nevertheless, the neuroprotective properties of NMNATs in the central nervous system have not yet been well studied, and its potential role in the injured and healthy immature brain is even less well understood. The goal of this project is to test the hypothesis that endogenous and exogenous NMNAT proteins protect developing neurons from the effects of H-I by increasing the calcium buffering capacity of mitochondria. In order to test this hypothesis, this proposal will utilized various biochemical, immunological, molecular and genetic techniques to assess the role of these proteins in injured developing neurons through the following aims: 1) To characterize the expression and role of endogenous NMNATs in cell death following neonatal H-I utilizing immunochemical and bimolecular methods of NMNAT gene expression and employing gene knock-down techniques. 2) To assess the neuroprotective role of exogenous NMNAT 1, 2 and 3 in the neonatal brain following H-I using transgenic animal models and gene transduction systems. 3) Investigate the neuroprotective mechanism of NMNAT(s) in the neonatal brain employing biochemical and bioluminescent techniques to examine mitochondrial physiology in over- and under-expressing NMNAT in vitro models of H-I and excitotoxicity. The proposed experiments will likely shed new light in our understanding of the biological processes that regulate neuronal cell death in the developing newborn brain. Furthermore, this knowledge may ultimately lead to identifying novel therapeutic targets for the treatment and prevention of the neurological consequences attributed to brain injury in the newborn.

描述（由申请人提供）：这一指导职业发展奖的目标是促进Rafael Galindo博士向独立的过渡，因为研究受伤的神经元受伤的神经元死亡的神经生物学机制。拟议的实验将在David Holtzman博士的指导下进行，他们将研究一个NAD+代谢酶（称为烟酰胺单核苷酸腺苷腺苷转移酶（NMNATS））的神经保护作用，在新生脑脑缺氧 - 高素质 - Ischemia（H-i Ischemia）中。这种伤害形式是新生儿时期最常见的死亡原因，占围产期脑损伤的慢性神经系统疾病的很大一部分。尽管患病率很高，但有限的有限可用于暴露于H-I的新生儿的疗法。缺乏有效的临床干预措施部分归因于我们对调节健康和受伤的发育中大脑中神经元死亡途径的神经生物学机制和分子的不完全理解。 NMNAT是已证明具有强神经保护特性的分子，并且可能重要地参与了各种中枢和周围神经系统疾病的神经退行性机理。然而，NMNAT在中枢神经系统中的神经保护特性尚未得到很好的研究，其在受伤和健康的未成熟大脑中的潜在作用甚至不太了解。该项目的目的是检验以下假设：内源性和外源性NMNAT蛋白通过增加线粒体的钙缓冲能力来保护发育神经元免受H-I影响。 In order to test this hypothesis, this proposal will utilized various biochemical, immunological, molecular and genetic techniques to assess the role of these proteins in injured developing neurons through the following aims: 1) To characterize the expression and role of endogenous NMNATs in cell death following neonatal H-I utilizing immunochemical and bimolecular methods of NMNAT gene expression and employing gene knock-down技术。 2）使用转基因动物模型和基因转导系统，在H-I后，在新生儿大脑中评估新生儿大脑中外源性NMNAT 1、2和3的神经保护作用。 3）研究NMNAT在新生儿大脑中的神经保护机制，采用生物化学和生物发光技术在过度表达和表达的NMNAT的H-I模型中检查线粒体生理的H-I和兴奋性。提出的实验可能会在我们对调节新生儿神经元细胞死亡的生物学过程的理解中发明新的启示。此外，这些知识最终可能导致鉴定出新的治疗靶标，以治疗和预防新生儿脑损伤的神经系统后果。