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.

项目总结/摘要 溶质载体，包括主要易化因子超家族（MFS）的成员，包括一个大的和 一组未充分研究的蛋白质，在不同的生物过程中发挥作用，包括金属运输和压力 阻力以酿酒酵母为模式生物，我们以前的研究表明， 临床上有前途的抗癌钌络合物KP 1019诱导进化上保守的 MFS蛋白Tpo 1.然而，驱动这种诱导的机制尚不清楚。 鉴于Tpo 1与许多其他MFS蛋白一样，从细胞中排出多种毒素，KP 1019 缺乏TPO 1的酵母的抗性是违反直觉的。对这种令人惊讶的表型的可能解释 包括其它药物转运蛋白补偿性激活或多胺稳态的扰动， 已知由TPO 1调节。为了推进我们的长期目标，确定如何溶质载体，包括 它们的调节，调节KP 1019耐受性，我们将追求三个具体目标，重点是确定 KP 1019和Tpo 1之间关系的上游调节子和下游效应子。我们特别 建议1）确定KP 1019诱导TPO 1的机制和生理意义，2） 表征由TPO 1缺失引起的KP 1019抗性，和3）发现KP 1019的新调节剂 电阻/灵敏度。 删除我们先前研究中涉及的转录因子基因将能够鉴定 负责KP 1019的药物依赖性诱导的调节剂。转录组学分析将为 假设驱动的实验，旨在确定药物外排的代偿性激活的作用， 缺乏TPO 1的酵母对KP 1019抗性中的多胺稳态。一个定量的表型筛选， 酵母缺失收集将有助于发现新的KP 1019耐受性调节剂， 建立预测患者对抗癌钌络合物反应的遗传标记。 此外，该项目将加强弗曼大学的研究基础设施，创造新的 本科生有机会参与高通量技术与假设驱动的配对 实验，这将增加他们追求生物医学科学事业的可能性的组合。