Novel single-cell mass spectrometry methods to assess the role of intracellular drug concentration and metabolism in antimicrobial treatment failure

评估细胞内药物浓度和代谢在抗菌治疗失败中的作用的新型单细胞质谱方法

• 批准号: 10714351
• 负责人: Laura-Isobel McCall
• 金额: --
• 依托单位: UNIVERSITY OF OKLAHOMA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2023-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10714351
• 关键词: 2019-nCoV; Address; Anti-Bacterial Agents; Antineoplastic Agents; Antiparasitic Agents; Antiviral Agents; Bacterial Infections; Benznidazole; Biohazardous Substance; Biological; Biological Availability; Cells; Cellular Metabolic Process; Cessation of life; Characteristics; Communicable Diseases; Communities; Development; Dose; Drug Kinetics; Drug resistance; Ensure; Environment; Failure; Goals; Health; Heterogeneity; In Vitro; Individual; Infection; Link; Malignant Neoplasms; Mass Spectrum Analysis; Metabolic; Metabolism; Methodology; Methods; Microbial Drug Resistance; Mission; Nifurtimox; Parasites; Parasitic infection; Parents; Penetration; Pharmaceutical Preparations; Prodrugs; Public Health; Research; Role; Spectrometry; Streptococcus pyogenes; System; Techniques; Technology; Testing; Tissues; Treatment Failure; Trypanosoma cruzi; United States National Institutes of Health; Virus Diseases; Work; analytical method; antimicrobial; antimicrobial drug; clinical implementation; drug development; drug metabolism; drug testing; efflux pump; fexinidazole; improved; infectious disease treatment; innovation; insight; instrument; metabolic abnormality assessment; mortality; neoplastic cell; novel; pathogen; success; technology/technique; uptake

Project Summary/Abstract Infectious disease treatment failure is a critical health issue. It is predicted to become the leading cause of mortality by 2050, superseding cancer. Treatment failure can be caused by a combination of factors, including active antimicrobial drug resistance, for example via efflux pumps, and pathogen dormancy, tolerance and persistence mechanisms. However, these factors cannot alone account for all cases of treatment failure. Indeed, bioavailability and pharmacokinetics are the 3rd most common cause of failure during drug development. Pharmacokinetic studies, however, have predominantly focused on biofluid and tissue drug levels. This is insufficient in the case of intracellular pathogens, where intracellular drug accumulation is critical to ensure pathogen clearance. Single-cell mass spectrometry (SCMS) studies in the field of cancer drug development have revealed extensive heterogeneity in intracellular drug levels. However, until now, the technology and techniques to safely perform such analyses in the context of intracellular pathogen infection have been lacking. This is a critical gap in our ability to understand infectious disease treatment failure and to guide infectious disease drug development. To address this gap, proposal MPIs have developed a biosafety- compatible SCMS method to quantify intracellular drug and metabolite levels in the context of Trypanosoma cruzi parasite infection. The overall objective of this proposal is to demonstrate the broad applicability of this method to determine the relationship between lower intracellular drug levels and drug metabolism vs failure to clear intracellular pathogens. The central hypothesis of this proposal is that heterogenous intracellular drug levels and intracellular drug metabolism is a key contributor to antimicrobial treatment failure, and that this mechanism can be revealed using novel SCMS approaches with broad applicability. We will test this central hypothesis using three complementary yet independent aims. Aim 1 will use a Trypanosoma cruzi parasite infection system to relate heterogenous antiparasitic drug levels between cells, to in vitro T. cruzi treatment failure. Aim 2 will use the same T. cruzi infection system to relate heterogenous antiparasitic drug metabolism between cells, to in vitro T. cruzi treatment failure. Aim 3 will broaden the applicability of these SCMS techniques to quantify specific antibacterials and antivirals, and to relate their levels to treatment failure in the context of bacterial and viral infection. The proposed research is innovative because it will lead to the development of a new bioanalytical technology to address the critical biological problem of antimicrobial treatment failure. The proposed research is significant because it will lead to an expanded understanding of the mechanisms of treatment failure in infectious diseases.

项目摘要/摘要 传染病治疗失败是一个关键的健康问题。预计将成为 到2050年，死亡率取代了癌症。治疗失败可能是由多种因素造成的，包括 主动抗菌耐药性，例如通过外排泵以及病原体的休眠，耐受性和 持久机制。但是，这些因素不能单独解释所有治疗失败的情况。 实际上，生物利用度和药代动力学是药物期间第三大最常见的失败原因 发展。但是，药代动力学研究主要集中在生物流体和组织药物上 水平。在细胞内病原体的情况下，这是不够的，那里细胞内药物的积累至关重要 确保病原体清除率。癌症药物领域的单细胞质谱法（SCM）研究 发育已经显示出细胞内药物水平的广泛异质性。但是，直到现在， 在细胞内病原体感染的背景下，可以安全执行此类分析的技术和技术 缺乏。这是我们了解传染病治疗失败和 指导传染病药物开发。为了解决这一差距，MPI的提案已经开发了一个生物安全 - 在锥虫瘤中量化细胞内药物和代谢物水平的兼容SCMS方法 克鲁兹寄生虫感染。该提案的总体目的是证明这一点的广泛适用性 确定较低细胞内药物水平与药物代谢与未能失败之间关系的方法 清除细胞内病原体。该提议的中心假设是异质细胞内药物 水平和细胞内药物代谢是抗菌治疗衰竭的关键因素，这是 可以使用具有广泛适用性的新型SCMS方法来揭示机制。我们将测试这个中央 假设使用三个互补但独立的目标。 AIM 1将使用Cruzi寄生虫锥虫瘤 感染系统将细胞之间的异质抗寄生虫药物水平与体外T. Cruzi治疗相关联 失败。 AIM 2将使用相同的克鲁兹感染系统与异质抗寄生虫药物代谢相关联 在细胞之间，进行体外Cruzi治疗衰竭。 AIM 3将扩大这些SCM的适用性 量化特定抗菌和抗病毒药的技术，并将其水平与治疗失败相关联 细菌和病毒感染的背景。拟议的研究具有创新性，因为它将导致 开发一种新的生物分析技术来解决抗菌剂的关键生物学问题 治疗失败。拟议的研究很重要，因为它将导致对 传染病的治疗失败机制。