Optical platform for functional longitudinal imaging of metabolite uptake in vivo

用于体内代谢物摄取功能纵向成像的光学平台

• 批准号: 10585764
• 负责人: Elena Goun
• 金额: $ 34.2万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-15 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10585764
• 关键词: Ablation; Address; Affinity; Amino Acids; Animal Cancer Model; Animal Disease Models; Animal Model; Animals; Azides; Behavior; Biological; Biological Assay; Breast Cancer Model; CRISPR/Cas technology; Categories; Cell Culture Techniques; Cell model; Cells; Chemistry; Clinical; Dependence; Development; Diabetes Mellitus; Disease Progression; Enzymes; Evaluation; Fatty Acids; Fireflies; Generations; Glucose; Glutamine; Goals; Human; Human Pathology; Image; Imaging Device; Imaging Techniques; In Vitro; Intervention; Intraperitoneal Injections; Investigation; Isotopes; Kinetics; Ligation; Light; Luc Gene; Luciferases; Magnetic Resonance Imaging; Malignant Neoplasms; Measurement; Metabolic; Metabolic Diseases; Metabolism; Methods; Modification; Molecular Probes; Monitor; Mus; Nature; Neurodegenerative Disorders; Normal Cell; Nucleosides; Nucleotides; Nutrient; Oleic Acids; Oncology; Optics; Oral; Pathologic; Physiological; Play; Positron-Emission Tomography; Process; Production; Proliferating; Publishing; Radioactive; Reaction; Reagent; Reporting; Role; Route; Safety; Signal Transduction; Specificity; Structure; Synthesis Chemistry; System; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Toxic effect; Transgenic Mice; Transgenic Organisms; Tryptophan; Tube; Validation; Work; absorption; analytical method; analytical tool; biological systems; bioluminescence imaging; cancer cell; cancer therapy; chemotherapy; clinical application; cost; design; drug discovery; effective therapy; experimental study; glucose uptake; human model; imaging modality; imaging platform; improved; in vitro Model; in vivo; inhibitor; interest; intravenous injection; luciferin; metabolic abnormality assessment; molecular imaging; new technology; non-invasive imaging; nonalcoholic steatohepatitis; novel; nuclear imaging; optical imaging; serial imaging; simulation; single photon emission computed tomography; small hairpin RNA; success; therapeutic target; tool; tumor; tumor growth; tumor metabolism; tumor microenvironment; tumorigenesis; uptake

PROJECT SUMMARY Cancer cells undergo metabolic reprogramming in order to meet elevated energy requirements to fuel proliferation, thus resulting in their differential utilization of many essential metabolites compared to normal cells. Recent advancements in the field of cancer metabolic reprogramming demonstrated significant increase in efficiency of standard cancer treatments when combined with cancer metabolic inhibitors. However, tumor metabolic reprogramming remains poorly understood for the majority of cancers. Moreover, many recent reports revealed evidence that the metabolism of cancer cells in vitro can differ significantly from that of in vivo because in vitro models lack complexity of the tumor microenvironment. However, the progress of studying tumor metabolism in vivo is significantly hampered by the lack of efficient tools that allow real-time noninvasive imaging and quantification of metabolite absorption in animal models of cancer which closely reflect human pathologies. Current strategies have significant limitations and mostly rely on MRI, nuclear imaging techniques such as PET/SPECT, and endpoint ex vivo quantification of metabolite absorption (ex. MS). Here, we propose to develop a novel optical imaging platform that has several important advantages over the existing methods, and allows noninvasive evaluation of the uptake of several essential metabolites using highly sensitive and quantifiable bioluminescent imaging. The method is independent of radioactive and/or short-lived isotopes, less costly, and allows longitudinal monitoring of metabolite absorption during disease progression (e.g., cancer development or clinical intervention such as chemotherapy). While the first application of this approach has been already successfully validated by us using glucose as an example (Maric et.al., Nat Methods, 2019), we propose to expand this technology to develop novel probes to study uptake of several amino acids, fatty acids, and nucleosides that all play central role in cancer metabolic reprogramming. We will perform thorough validation of this platform in cells, healthy transgenic mice and murine animal cancer models to assure that the reagents fulfill the requirements for physiological behavior, stability, safety, and robust signal generation both in vitro and in vivo. In addition, we will optimize in vivo delivery routes, vehicles, and concentrations to achieve high signal/background ratios. In summary, the overall goal of this study is to generate a novel optical imaging platform that would become a universal analytical tool for monitoring nutrient uptake in live cells and animal models of disease. While we plan to apply this platform to unravel tumor metabolic reprogramming, the same method could be adapted for studies of several other important human pathologies, in which changes in metabolism are known to play a significant role, such as diabetes, neurodegenerative diseases, nonalcoholic steatohepatitis (NASH), and many others. Therefore, this novel technology is expected to have a strong, enabling, and long-lasting impact on many physiological and pathological investigations in the field of metabolism and will become a valuable tool for drug discovery, applicable to oncology and other metabolic disorders.

项目摘要 癌细胞进行代谢重编程，以满足提高的能量需求， 增殖，从而导致它们与正常细胞相比对许多必需代谢物的差异利用。 癌症代谢重编程领域的最新进展表明， 当与癌症代谢抑制剂组合时，标准癌症治疗的效率。然而，肿瘤 对于大多数癌症，代谢重编程仍然知之甚少。此外，最近许多报告 揭示的证据表明，癌细胞在体外的代谢可以显着不同于在体内，因为 体外模型缺乏肿瘤微环境的复杂性。然而，研究肿瘤的进展 由于缺乏允许实时非侵入性成像的有效工具， 以及在密切反映人类病理学的癌症动物模型中定量代谢物吸收。 目前的策略有很大的局限性，主要依赖于MRI，核成像技术，如 PET/SPECT和代谢物吸收的终点离体定量（例如，MS）。在此，我们建议开发 一种新颖的光学成像平台，其具有优于现有方法的几个重要优点，并且允许 使用高灵敏度和可定量的非侵入性评估几种必需代谢物的摄取 生物发光成像该方法不依赖于放射性和/或短寿命同位素，成本较低， 允许在疾病进展期间纵向监测代谢物吸收（例如，癌症发展或 临床干预如化疗）。虽然这种方法的第一次应用已经 我们使用葡萄糖作为例子成功地验证了这一点（Maric等人，Nat Methods，2019），我们建议 扩展这项技术，开发新的探针，研究几种氨基酸，脂肪酸和 在癌症代谢重编程中发挥核心作用的核苷。我们将进行彻底的验证， 该平台在细胞、健康转基因小鼠和鼠动物癌症模型中的应用，以确保试剂满足 对体外和体内生理行为、稳定性、安全性和稳健信号生成的要求 vivo.此外，我们还将优化体内递送途径、载体和浓度，以实现高浓度的药物递送。 信号/背景比。综上所述，本研究的总体目标是产生一种新颖的光学成像平台 这将成为一个通用的分析工具，用于监测活细胞和动物模型的营养吸收， 疾病虽然我们计划应用这个平台来解开肿瘤代谢重编程，但同样的方法可以 适用于其他几种重要的人类病理学的研究，其中已知代谢的变化 发挥重要作用，如糖尿病、神经退行性疾病、非酒精性脂肪性肝炎（NASH）， 和其他许多人.因此，这项新颖的技术有望产生强大、有利和持久的影响 在代谢领域的许多生理和病理调查，并将成为一个有价值的工具 用于药物发现，适用于肿瘤学和其他代谢疾病。