Mechanisms of elesclomol-mediated copper delivery to cuproenzymes in cells

来氯醇介导的铜向细胞内铜酶的传递机制

基本信息

批准号：
10468271
负责人：
Vishal Mahendrasingh Gohil
金额：
$ 30.86万
依托单位：
TEXAS A&M AGRILIFE RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10468271
关键词：
Affect Affinity Animal Model Animals Antineoplastic Agents Behavioral Binding Biochemical Biochemical Genetics Biochemical Process Biogenesis Biological Biological Models Brain Cancer Patient Carrier Proteins Cells Cellular Membrane Child Complex Connective Tissue Copper Defect Development Disease Dose Enzymes Exhibits Free Radicals Functional disorder Genes Genetic Goals Growth Growth and Development function Human Impairment In Vitro Iron Location Malignant Neoplasms Mammalian Cell Mediating Membrane Menkes Kinky Hair Syndrome Metabolism Micronutrients Mitochondria Mitochondrial Diseases Modeling Mus Mutation Organelles Oxidases Pathology Pathway interactions Pharmacology Pharmacotherapy Phenotype Physiological Processes Physiology Positioning Attribute Production Regimen Research Respiratory Chain Role Salts Site Specificity Sulfur Symptoms Testing Therapeutic Toxic effect Trace metal Transition Elements Work Yeasts Zebrafish absorption base body system cell injury cofactor cytochrome c oxidase dietary disease phenotype effective therapy genetic approach human model hydrophilicity hypocupremia infancy loss of function mutation mortality mouse model nanomolar neuropathology neurotransmitter biosynthesis oxidative damage prevent

项目摘要

PROJECT SUMMARY Copper (Cu) is an essential micronutrient that is required for growth and development of humans. The requirement of Cu is attributed to its role as a catalytic cofactor for a number of enzymes involved in various physiological processes, including mitochondrial energy production, neurotransmitter biosynthesis, and connective tissue formation. Owing to its diverse roles in cellular and organismal physiology, loss-of-function mutations in genes required for Cu absorption or transport to cuproenzymes often result in fatal infantile disorders. For example, defects in the steps involved in the mitochondrial Cu delivery pathway to cytochrome c oxidase (CcO), a mitochondrial cuproenzyme, are associated with a subset of mitochondrial disorders, while disruption in delivering dietary Cu to different organ systems results in Menkes disease. Currently, therapies for these diseases are experimental and largely ineffective at preventing early mortality. To identify compounds that can deliver Cu to cuproenzymes and thus overcome these defects, we performed a targeted screen for Cu-binding pharmacological agents and identified elesclomol (ES) as a potent Cu-transporting molecule. Our initial studies have shown that low nanomolar concentrations of ES can deliver Cu to CcO in yeast, zebrafish, and mouse models of Cu deficiency. Having made this breakthrough, we are now in position to test our hypothesis that ES, which was originally developed as a cancer drug, can be repurposed to safely deliver Cu to different cuproenzymes and rescue disease phenotypes associated with Cu deficiency disorders. To do this, we will first determine where and how ES releases Cu inside cells and whether this can be achieved without damaging cells. Second, we will utilize our newly constructed zebrafish models with specific defects in systemic or mitochondrial Cu metabolism and a mouse model of Menkes disease to test the efficiency of ES in rescuing mitochondrial, cellular, and organismal phenotypes by delivering Cu to cuproenzymes. Thus, the proposed work will uncover the mechanism of ES-mediated intracellular Cu transport and advance a promising therapeutic molecule for the treatment of debilitating and frequently fatal Cu deficiency disorders.

项目摘要铜（CU）是人类生长和发展所需的必不可少的微量营养素。这 Cu的要求归因于其作为涉及各种酶的催化辅因子的作用生理过程，包括线粒体能量产生，神经递质生物合成和结缔组织形成。由于其在细胞和生物生理学中的各种作用，功能丧失铜吸收或运输到库糖酶所需的基因突变通常会导致致命的婴儿疾病。例如，线粒体Cu递送途径与细胞色素c的缺陷中的缺陷氧化酶（CCO）是一种线粒体cupro酶，与线粒体疾病的一部分有关将饮食CU传递给不同器官系统的破坏会导致Menkes疾病。目前，用于治疗这些疾病是实验性的，在预防早期死亡率方面基本上无效。识别化合物可以将Cu输送到库酶，从而克服这些缺陷，我们进行了一个针对性的屏幕 Cu结合药理学剂，并将Elesclomol（ES）鉴定为有效的Cu传输分子。我们的初步研究表明，低纳摩尔浓度的ES可以在斑马鱼酵母中向CCO递送CU，和Cu缺乏的鼠标模型。实现了这一突破后，我们现在可以测试我们的可以重新使用的假设，即最初作为癌症药物开发的ES可以安全地提供Cu 与与CU缺乏障碍有关的不同库酶和救援疾病表型。为此，我们将首先确定ES在细胞内部的何处以及如何释放Cu，以及是否可以实现这一点破坏性细胞。其次，我们将利用具有特定缺陷的新建斑马鱼模型系统性或线粒体Cu代谢和Menkes疾病的小鼠模型，以测试ES的效率通过将Cu传递到鸡冠酶来营救线粒体，细胞和有机表型。因此，拟议的工作将揭示ES介导的细胞内Cu运输的机制，并提高有前途的治疗衰弱和经常致命的Cu缺乏症的治疗分子。