Cellular Basis for the Antifungal Activity of Amiodarone

胺碘酮抗真菌活性的细胞基础

基本信息

批准号：
7570031
负责人：
RAJINI RAO
金额：
$ 41.63万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-03-01 至 2011-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7570031
关键词：
Amiodarone Amphotericin B Anabolism Animal Model Anti-Arrhythmia Agents Antifungal Agents Antioxidants Apoptosis Apoptotic Appearance Aspergillus Aspergillus fumigatus Atrial Fibrillation Biochemical Biochemical Genetics Biological Biological Assay Calcineurin Calcium Candida Candida albicans Caspase Caspase Inhibitor Caspofungin Cell Death Cell membrane Cells Chelating Agents Chemicals Collection Combined Modality Therapy Cryptococcus Cryptococcus neoformans DNA Microarray Chip Data Databases Defect Development Disseminated candidiasis Dose Drug Delivery Systems Drug Design Drug resistance Drug toxicity Drug usage Ergosterol Event Exhibits Fluconazole Fungal Drug Resistance Generations Genes Goals Growth Homeostasis Hypersensitivity In Vitro Industrial fungicide Intervention Ion Channel Itraconazole Knock-out Lead Libraries Link Mediating Miconazole Microarray Analysis Mitochondria Modeling Molecular Molecular Profiling Mus Mutation Mycoses Pathway interactions Pharmaceutical Preparations Proteins Reactive Oxygen Species Research Personnel Resistance Reverse Transcriptase Polymerase Chain Reaction Saccharomyces Saccharomyces cerevisiae Saccharomycetales Serine Signal Pathway Signaling Pathway Gene Testing Time Toxic effect Vacuole Ventricular Arrhythmia Work Yeasts analog base cytochrome c dronedarone drug discovery fungus genome-wide in vivo inhibitor/antagonist knockout gene mitochondrial membrane mutant novel pathogen programs public health relevance research study response synergism trafficking weapons

项目摘要

The goal of this proposal is to elucidate the pathway of programmed cell death by a novel antifungal agent and explore its use as an antimycotic adjunct. Amiodarone is an effective antiarrhythmic drug that was recently discovered to have potent and broad range fungicidal activity. We have shown that amiodarone toxicity in the yeast Saccharomyces cerevisiae is mediated by disruption of calcium homeostasis, followed by the appearance of apoptosis markers and cell death. In Aim 1, we propose to use a combination of biochemical and cell biological approaches to determine the identity and temporal order of events leading from the initial burst of cytosolic calcium to cell death. We will seek to validate key findings in the pathogenic yeast Candida albicans as proof-of-principle for the universality of the fungicidal mechanism of amiodarone. In Aim 2, we will use a variety of genome-wide approaches to identify the genes and signaling pathways that contribute to amiodarone-induced cell death. Genes identified by these studies will be organized into biomodules and placed in cellular pathways by integrating experimental data from high-throughput biochemical assays with information from online databases to give a global view of drug toxicity. We have shown that low doses of amiodarone exhibit potent synergism with existing antifungals against pathogenic fungal species of Candida, Cryptococcus and Aspergillus. Drug synergy from these in vitro studies will form the basis for combination therapy that will be explored in a murine model of systemic Candidiasis (Aim3). Taken together these studies will develop the calcium-mediated cell death pathway as a major new drug discovery target opportunity. Amiodarone will serve as a model test compound for targeting this pathway and for validating the potential of a new class of antifungal potentiating agents. The public health relevance of this project arises from the emergence of new fungal pathogens and drug resistant fungi, and the urgent need for alternative strategies in the management of fungal infections.

该提案的目的是阐明新型抗真菌药物的编程细胞死亡途径并探索其用作抗菌毛辅助物。胺碘酮是一种有效的抗心律失常药物最近被发现具有有效且范围广泛的杀真菌活动。我们已经证明了胺碘酮酿酒酵母的酵母糖粉中的毒性是通过钙稳态的破坏来介导的，随后是通过凋亡标记和细胞死亡的出现。在AIM 1中，我们建议将生化和细胞生物学方法来确定事件的身份和时间顺序从胞质钙的初始爆发到细胞死亡。我们将寻求验证致病性的关键发现酵母白色念珠菌作为胺碘酮杀真菌机制普遍性的原理证明。在AIM 2中，我们将使用各种全基因组方法来识别基因和信号通路有助于胺碘酮诱导的细胞死亡。这些研究确定的基因将被组织到生物模型并通过整合高通量的实验数据放置在细胞途径中来自在线数据库中的信息的生化测定法，以全球性观察药物毒性。我们有表明低剂量的氨二酮与现有的抗真菌抗体表现出有效的协同作用真菌的念珠菌，加密环球和曲曲霉。这些体外研究的药物协同作用将形成联合疗法的基础将在系统性念珠菌病的鼠模型中探索（AIM3）。综上所述，这些研究将发展为钙介导的细胞死亡途径作为主要的新药发现目标机会。胺碘酮将用作针对此途径的模型测试化合物，验证新类抗真菌增强剂的潜力。该项目的公共卫生相关性来自新真菌病原体和药物的出现抗真菌，迫切需要在真菌感染管理中采取替代策略。