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.

项目概要/摘要 溶质载体，其中包括主要促进子超家族 (MFS) 的成员，由一个大型且 正在研究的一组在不同生物过程中发挥作用的蛋白质，包括金属运输和应激 反抗。我们之前的研究表明，以酿酒酵母为模式生物 临床上有前景的抗癌钌复合物 KP1019 诱导进化保守的表达 MFS 蛋白 Tpo1。然而，驱动这种诱导的机制尚不清楚。 鉴于 Tpo1 与许多其他 MFS 蛋白一样，可从细胞中排出多种毒素，因此 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代偿性激活或多胺稳态的扰动，这是 已知受 Tpo1 调节。为了推进我们确定溶质载体的长期目标，包括 他们的监管，调节 KP1019 耐受性，我们将追求三个具体目标，重点是确定 KP1019和Tpo1之间关系的上游调节器和下游效应器。具体来说，我们 建议 1) 确定 KP1019 诱导 TPO1 的机制和生理意义，2) 表征由 TPO1 缺失引起的 KP1019 抗性，以及 3) 发现 KP1019 的新型调节剂 电阻/灵敏度。 删除我们之前研究中涉及的转录因子基因将能够识别 负责 KP1019 药物依赖性诱导的调节器。转录组分析将告知 假设驱动的实验旨在确定药物流出和补偿激活的作用 缺乏 TPO1 的酵母 KP1019 抗性中的多胺稳态。定量表组筛选 酵母删除集合将有助于发现 KP1019 耐受性的新调节剂，从而使 建立预测患者对抗癌钌络合物反应的遗传标记。 此外，该项目将增强弗曼大学的研究基础设施，创造新的 本科生有机会接触高通量技术以及假设驱动的技术 实验，这两者的结合将增加他们从事生物医学科学职业的可能性。