Inborn Errors of Long Chain Fat Metabolism

长链脂肪代谢先天性错误

基本信息

批准号：
7810760
负责人：
GERARD VOCKLEY
金额：
$ 43.9万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-20 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7810760
关键词：
Acute Liver Failure Acyl CoA Dehydrogenases Acyl Coenzyme A Address Affect Alternative Splicing Amino Acid Motifs Amino Acids Animals Bile Acid Biosynthesis Pathway Bile Acids Biochemical Biogenesis Cardiomyopathies Catabolism Cataloging Catalogs Cell Fractionation Cell Nucleus Cells Cessation of life Chemicals Chenodeoxycholic Acid Clinical Complex Data Defect Development Disease Enzymes Equilibrium Esters Family Fatty Acids Fatty Liver Functional disorder Funding Gene Family Generations Genes Genetic Genome Genotype Goals Hereditary Disease Histologic Human Immunohistochemistry Inborn Errors of Metabolism Inborn Genetic Diseases Inner mitochondrial membrane Institutes Knockout Mice Lead Learning Lecithin Length Lipids Liver Failure Location Long-Chain-Acyl-CoA Dehydrogenase Lung Medicine Metabolic Metabolism Mitochondria Mitochondrial Matrix Modeling Molecular Molecular Conformation Mus Mutation Newborn Infant Pathway interactions Patients Pattern Phenotype Physiological Premature Infant Preparation Process Proteins Recovery Rest Reye Syndrome Role Sampling Structure-Activity Relationship Substrate Specificity Surveys Testing Texas Time Tissues United States National Institutes of Health Variant acyl-CoA dehydrogenase base dehydrogenation enzyme deficiency fatty acid oxidation in vivo lipid metabolism member mouse model mutant novel oxidation public health relevance surfactant tissue/cell culture

项目摘要

DESCRIPTION (provided by applicant): This application addresses Notice Number (NOT-OD-09-058) and Notice Title: NIH Announces the Availability of Recovery Act Funds for Competitive Revision Applications, and asks for competitive revision of R01DK78755. The acyl-CoA dehydrogenases (ACDs) are a family of multimeric flavoenzymes that catalyze the 1,2-dehydrogenation of acyl-CoA esters in fatty acid 2-oxidation and amino acid catabolism. Inborn errors of metabolism have been identified in seven of the ACDs. The long range objective of this project has been to investigate important structure/function relationships in the ACD gene family. We have described and characterized several new members of the ACD gene family. Among these are 3 enzymes with significant activities with long chain substrates: long and very long chain acyl-CoA dehydrogenases (LCAD and VLCAD, respectively), and ACD9 and. Our prior and preliminary studies show that these enzymes have distinct substrate utilization profiles, tissue and developmental expression patterns, exist in multiple active forms in the cell, and are present in multiple subcellular locations. The goal of the funded application is to characterize the physiologic roles of LCAD, VLCAD, and ACD9 and explore the ramifications of genetic deficiencies of these enzymes in humans and mouse models. Specific Aim 2 of the original application is to more completely characterize ACD9 and its deficiency in humans. Specific Aim 2a is identification of additional patients with ACD9 deficiency and definition of its clinical spectrum. Specific Aim 2b is characterization of the subcellular distribution of ACD9 and the function and molecular configuration of ACD9 protein outside of mitochondria. I hypothesize that this alternative form of ACD9 has non-enzymatic "moonlighting" functions in the cell. I am requesting a supplement to the application to extend my ability to address these aims. First, I have identified more potential deficient patients than originally anticipated. I am requesting funds to support additional technician time to process these samples more quickly and accelerate our progress. Secondly, our understanding of the pathophysiology of ACAD9 deficiency and our ability to study the moonlighting functions of this gene product are limited by the availability of only tissue culture cells with ACAD9 deficiency. To augment these studies, I am requesting supplemental funds to purchase an ACAD9 knock out mouse from the Texas A&M Institute for Genome Medicine, which has it available in its catalogue. The biochemical and pathophysiologic phenotype of the animal will be characterized including metabolic profiling, complete histologic survey, and physiologic effects of the deficiency on the animal under basal and resting conditions. Immunohistochemistry and subcellular fractionation studies will examine for presences of ACAD9 in non-mitochondrial locations and loss in the knock out mouse model. PUBLIC HEALTH RELEVANCE: The acyl-CoA dehydrogenases are important enzymes in maintaining normal chemical balance in the body. We have identified a new genetic disorder of one of these enzymes that leads to liver failure. Studying this disorder is important to learn more about its clinical presentation and treatment

说明（由申请人提供）：本申请涉及通知编号 (NOT-OD-09-058) 和通知标题：NIH 宣布恢复法案资金可用于竞争性修订申请，并要求对 R01DK78755 进行竞争性修订。酰基辅酶 A 脱氢酶 (ACD) 是一个多聚黄素酶家族，可催化脂肪酸 2-氧化和氨基酸分解代谢中酰基辅酶 A 酯的 1,2-脱氢。在七种 ACD 中已发现先天性代谢缺陷。该项目的长期目标是研究 ACD 基因家族中重要的结构/功能关系。我们已经描述并表征了 ACD 基因家族的几个新成员。其中有 3 种酶对长链底物具有显着活性：长链和极长链酰基辅酶 A 脱氢酶（分别为 LCAD 和 VLCAD）以及 ACD9 和。我们之前和初步的研究表明，这些酶具有不同的底物利用概况、组织和发育表达模式，在细胞中以多种活性形式存在，并且存在于多个亚细胞位置。该资助申请的目标是表征 LCAD、VLCAD 和 ACD9 的生理作用，并探索这些酶的遗传缺陷在人类和小鼠模型中的影响。原始申请的具体目标 2 是更完整地表征 ACD9 及其在人类中的缺陷。具体目标 2a 是识别其他 ACD9 缺陷患者并定义其临床谱。具体目标 2b 是表征 ACD9 的亚细胞分布以及 ACD9 蛋白在线粒体外的功能和分子构型。我推测 ACD9 的这种替代形式在细胞中具有非酶促的“兼职”功能。我请求对申请进行补充，以扩展我实现这些目标的能力。首先，我发现了比最初预期更多的潜在缺陷患者。我请求资金来支持额外的技术人员时间，以更快地处理这些样本并加快我们的进度。其次，我们对 ACAD9 缺陷的病理生理学的理解以及我们研究该基因产物的兼职功能的能力受到仅具有 ACAD9 缺陷的组织培养细胞的可用性的限制。为了加强这些研究，我请求补充资金从德克萨斯 A&M 基因组医学研究所购买 ACAD9 敲除小鼠，该小鼠在其目录中可以找到。将表征动物的生化和病理生理表型，包括代谢谱、完整的组织学调查以及基础和静息条件下缺陷对动物的生理影响。免疫组织化学和亚细胞分级分离研究将检查 ACAD9 在非线粒体位置的存在以及敲除小鼠模型中的缺失。公众健康相关性：酰基辅酶A脱氢酶是维持体内正常化学平衡的重要酶。我们已经发现了其中一种酶的一种新的遗传性疾病，它会导致肝衰竭。研究这种疾病对于了解更多有关其临床表现和治疗的信息非常重要