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：
9102309
负责人：
GERARD VOCKLEY
金额：
$ 40.65万
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：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9102309
关键词：
Affect Amino Acids Architecture Binding Biology CRISPR/Cas technology Cell Culture Techniques Cell Line Cells Clinical Clinical Trials Coenzyme Q10 Complex Core Protein Defect Development Disease Electron Microscopy Electron Transport Electron Transport Complex III Energy Metabolism Enzymes Fatty Acids Functional disorder Funding Genes Goals Grant Imagery In Vitro Inborn Genetic Diseases Induced Mutation Inflammation Inflammatory Inner mitochondrial membrane Knowledge Lipids Liver Long-Chain-Acyl-CoA Dehydrogenase Lung Metabolic Metabolism Mitochondria Mitochondrial Diseases Molecular Molecular Models Mutate Mutation Neonatal Screening Pathway interactions Patients Peptides Pharmacologic Substance Phenotype Positioning Attribute Process Proteins Proteomics Riboflavin Role Set protein Structure Supplementation System Techniques Therapeutic base chaperonin fatty acid oxidation insight lipid metabolism long chain fatty acid metabolic abnormality assessment molecular modeling mouse model mutant novel novel therapeutics oxidation pneumocyte protein complex protein function protein structure function public health relevance reconstruction research study small molecule small molecule therapeutics surfactant three dimensional structure

项目摘要

DESCRIPTION (provided by applicant) Disorders of mitochondrial (FAO) are among the most frequent identified through newborn screening in the US. FAO is traditionally viewed as an energy-generating, catabolic pathway but intermediates of this pathway can serve as key substrates for synthesis of other complex lipids. The long range objective of this project is to define the role of FAO proteins in intermediary metabolism and the clinical impact of their deficiency due to inborn errors. Progress made on these aims has provided revolutionary insight into the molecular architecture of mitochondrial energy metabolism and positions us to leverage this knowledge for the development of novel therapies for long chain fatty acid oxidation disorders. The overall goal of this renewal application is to use integrative biology to characterize the molecular architecture of mitochondrial energy metabolism and to understand the global metabolic defects induced by deficiency of single enzyme disorders. Specific Aim 1 is to identify the key protein interactions that stabilize the macromolecular mitochondrial energy complex. I will examine the interactions of three specific sets of proteins based on my previous findings Specific Aim 1a is to use proteomics techniques to examine amino acid residues critical to the formation and stability of the proteins of long chain FAO. Specific Aim 1b is to directly examine the three dimensional structure of the multifunctional energy complex. Specific Aim 2 is to examine the effects of mutations in long chain FAO proteins on the macromolecular mitochondrial energy complex. I hypothesize that some mutations will not only inactivate the mutated protein, but also disrupt the stability and function of the macromolecular mitochondrial energy complex Specific Aim 2a is to examine the effects of patient and CRISPR/Cas9 induced mutations in cell lines and an FAOD mouse model on interaction of the long chain FAO complex core proteins with their binding partners.. Specific Aim 2b is to examine the effects of mutations in ETFDH on its interaction with complex III. Specific Specific Aim 2c is to characterize new mutations in patients with FAODs. Aim 3 is to develop novel small molecule compounds to treat long chain FAODs. I have previously used molecular modeling and an in vitro cell system to identify potential therapeutic small molecule chaperonins and peptides that stabilizes the common Glu304Lys mutant MCAD protein, one of which is currently in clinical trials in patients. My collaborators and I have also shown that some ACAD9 mutations are stabilized by supplementation with excess riboflavin, similar to a finding with some ETFDH mutations. I hypothesize that additional therapeutic molecules have the potential to stabilize other mutant fatty acid oxidation proteins and mitigate the atypical inflammatory process seen in VLCAD deficiency. Specific Aim 3a is to examine the use of novel pharmaceutical agents known to affect energy metabolism and inflammation in cells from patients with FAODs. Specific Aim 3b is to examine the effect of these compounds in mouse models of FAODs in order to prepare for possible clinical trials in patients.

描述（由申请人提供）线粒体疾病 (FAO) 是美国新生儿筛查中最常见的疾病之一。FAO 传统上被视为一种能量生成、分解代谢途径，但该途径的中间体可以作为合成其他复杂脂质的关键底物。该项目的目的是确定FAO蛋白在中间代谢中的作用以及由于先天性错误而导致的FAO蛋白缺陷的临床影响，这些目标的进展为线粒体能量代谢的分子结构提供了革命性的见解，并使我们能够利用这些知识。该更新申请的总体目标是利用整合生物学来表征线粒体能量代谢的分子结构，并了解由单一酶紊乱引起的整体代谢缺陷。具体目标 1 是确定稳定大分子线粒体能量复合物的关键蛋白质相互作用。我将根据我之前的发现检查三组特定蛋白质的相互作用。具体目标 1a 是使用蛋白质组学技术来检查关键氨基酸残基。具体目标1b是直接检查多功能能量复合物的三维结构。具体目标2是检查长链FAO蛋白质的突变对大分子线粒体能量的影响。我认为一些突变不仅会使突变蛋白失活，还会破坏大分子线粒体能量复合物的稳定性和功能具体目标2a是检查患者和CRISPR/Cas9的影响。细胞系和FAOD小鼠模型中诱导突变对长链FAO复合物核心蛋白与其结合配偶体相互作用的影响。具体目标2b是检查ETFDH突变对其与复合物III相互作用的影响。目标 3 是开发新的小分子化合物来治疗长链FAODs，我之前使用分子模型和体外细胞系统来鉴定潜在的治疗性小分子伴侣蛋白和。稳定常见 Glu304Lys 突变体 MCAD 蛋白的肽，其中一种目前正在患者身上进行临床试验。研究表明，一些 ACAD9 突变可以通过补充过量核黄素来稳定，这与一些 ETFDH 突变的发现类似，主要是额外的治疗分子有可能稳定其他突变脂肪酸氧化蛋白，并减轻 VLCAD 缺乏症中出现的非典型炎症过程。具体目标 3a 是检查已知会影响 FAOD 患者细胞能量代谢和炎症的新型药物的使用情况。具体目标 3b 是检查这些化合物在小鼠模型中的作用。为可能的患者临床试验做准备。