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 溶质载体包括主要促进剂超家族(MFS)的成员，包括大的和 一组未被充分研究的蛋白质，它们在不同的生物过程中发挥作用，包括金属运输和应激 抵抗。以酿酒酵母为模式生物，我们之前的研究表明， 临床有前景的抗癌Ru络合物KP1019诱导进化保守基因的表达 MFS蛋白Tpo1。然而，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的酵母的抗药性是违反直觉的。对这种令人惊讶的表型的可能解释 包括其他药物转运体补偿性激活或多胺稳态的扰动，这是 已知受Tpo1调控。为了推进我们的长期目标，确定溶质载体，包括 他们的监管，调节KP1019的耐受性，我们将追求三个具体目标，重点是确定 KP1019与Tpo1关系的上游调控因子和下游效应因子。具体来说，我们 建议1)确定KP1019诱导Tpo1的机制(S)及其生理意义；2) 鉴定由TPO1缺失引起的KP1019抗性，以及3)发现KP1019的新调节子 电阻/灵敏度。 我们先前研究中涉及的转录因子基因的缺失将使我们能够识别 调节者S负责KP1019的药物依赖诱导。转录分析将提供信息 假说驱动的实验旨在确定药物外排和药物外排的代偿激活的作用 缺乏TPO1的酵母对KP1019抗性的多胺动态平衡。一种定量的表观筛选 酵母缺失收集将有助于发现KP1019耐受性的新调节子，使 预测患者对抗癌Ru络合物反应的遗传标记的建立。 此外，该项目将加强福尔曼大学的研究基础设施，创造新的 本科生接触高通量技术的机会与假设驱动的结合 实验，这种结合将增加他们在生物医学科学领域追求职业生涯的可能性。