Impact of Solute Carriers on Ruthenium Complex Sensitivity in Yeast

溶质载体对酵母中钌络合物敏感性的影响

• 批准号: 10360056
• 负责人: PAMELA HANSON
• 金额: $ 29.92万
• 依托单位: FURMAN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-03 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10360056
• 关键词: Acids; Animal Model; Antineoplastic Agents; Automobile Driving; Bacteria; Bacterial Infections; Bar Codes; Binding Sites; Biological Process; Buffers; Cancer Model; Cancer cell line; Carrier Proteins; Cell physiology; Cells; Clinical; Complement; Complex; Coupled; DNA Binding; DNA Damage; DNA Repair; Data; Drug Efflux; Drug resistance; Exposure to; Genetic; Genetic Markers; Goals; Herbicides; Homeostasis; Human; Indazoles; Light; Malignant Neoplasms; Mammals; Measures; Messenger RNA; Metals; Mutate; Mycoses; Nutrient; Organism; Orthologous Gene; Oxidative Stress; Pathway interactions; Pharmaceutical Preparations; Pharmacogenomics; Phenotype; Physiological; Plasma; Play; Polyamines; Proteins; Proteomics; Protozoan Infections; Publishing; Reactive Oxygen Species; Regulation; Research; Research Infrastructure; Resistance; Role; Ruthenium; Saccharomyces cerevisiae; Saccharomycetales; Science; Signal Transduction; Stress; Techniques; Testing; Toxic effect; Toxin; Transcription Factor AP-1; Transition Elements; Universities; Xenobiotics; Yeasts; anti-cancer; biological adaptation to stress; career; deletion library; drug sensitivity; efflux pump; emission spectrometry; experimental study; gene induction; genetic analysis; man; member; mutant; next generation sequencing; novel; patient response; phenomics; plant fungi; predictive marker; response; small molecule; solute; toxic metal; transcription factor; transcriptome sequencing; transcriptomics; undergraduate student; uptake

Project Summary/Abstract Solute carriers, which include members of the Major Facilitator Superfamily (MFS), comprise a large and understudied group of proteins with roles in diverse biological processes, including metal transport and stress resistance. Using Saccharomyces cerevisiae as a model organism, our previous studies showed that the clinically promising anticancer ruthenium complex KP1019 induces expression of the evolutionarily conserved MFS protein Tpo1. However, the mechanisms driving this induction are unknown. Given that Tpo1, like many other MFS proteins, effluxes a diverse range of toxins from cells, the KP1019 resistance of yeast lacking TPO1 is counterintuitive. Possible explanations for this surprising phenotype include compensatory activation of other drug transporters or perturbation of polyamine homeostasis, which is known to be regulated by Tpo1. To advance our long-term goal of determining how solute carriers, including their regulation, modulate KP1019 tolerance, we will pursue three specific aims focused on identifying the upstream regulators and downstream effectors of the relationship between KP1019 and Tpo1. Specifically, we propose to 1) determine the mechanism(s) and physiological significance of TPO1 induction by KP1019, 2) characterize the KP1019 resistance caused by deletion of TPO1, and 3) discover novel modulators of KP1019 resistance/sensitivity. Deletion of transcription factor genes implicated in our previous studies will enable identification the regulator(s) responsible for drug-dependent induction of KP1019. Transcriptomic analyses will inform hypothesis-driven experiments aimed at determining the roles of compensatory activation of drug efflux and polyamine homeostasis in the KP1019 resistance of yeast lacking TPO1. A quantitative phenomic screen of the yeast deletion collection will aid in discovering new modulators of KP1019 tolerance, enabling establishment of genetic markers that predict patient response to anticancer ruthenium complexes. Furthermore, this project will enhance the research infrastructure at Furman University, creating new opportunities for undergraduates to engage with high throughput techniques paired with hypothesis-driven experiments, a combination that will increase their likelihood of pursuing careers in the biomedical sciences.

项目摘要/摘要 包括主要促进者超级家族（MFS）的溶质载体，包括一个大型和 研究的一组蛋白质组在多种生物过程中，包括金属运输和应力 反抗。我们以前的研究使用酿酒酵母作为模型生物，表明 临床上有希望的抗癌ruthenium复合物kp1019诱导了进化保守的表达 MFS蛋白TPO1。但是，驱动这种诱导的机制尚不清楚。 鉴于TPO1像许多其他MFS蛋白一样，从细胞中排出各种毒素，KP1019 缺乏TPO1的酵母的抗性是违反直觉的。这种令人惊讶的表型的可能解释 包括其他药物转运蛋白的补偿性激活或多胺稳态的扰动，这是 已知由TPO1调节。促进我们的长期目标，即确定求解器如何，包括 他们的法规调节KP1019的容忍度，我们将追求三个专注于确定的特定目标 KP1019和TPO1之间关系的上游调节器和下游效应子。具体来说，我们 提议1）确定KP1019，2的TPO1诱导的机制和生理意义 表征由TPO1删除引起的KP1019抗性，3）发现KP1019的新型调节剂 电阻/灵敏度。 与我们先前研究有关的转录因子基因的缺失将使鉴定 负责药物依赖性KP1019的调节剂。转录组分析将告知 假设驱动的实验旨在确定药物外排和 缺乏TPO1的KP1019抗性的多胺稳态。定量现象屏幕 酵母删除收集将有助于发现KP1019耐受性的新调节剂，从而实现 建立遗传标志物，以预测患者对抗癌氟芬氏菌复合物的反应。 此外，该项目将增强弗曼大学的研究基础设施，创建新的 本科生与高吞吐量技术与假设驱动的机会 实验，这种组合将增加他们在生物医学科学中从事职业的可能性。