Metabolomics Core

代谢组学核心

• 批准号: 10487268
• 负责人: Daniel Crooks
• 金额: $ 235.68万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10487268
• 关键词: Abdomen; Aftercare; Algorithms; Animals; Bacteria; Biochemical; Biological; Biological Markers; Biopsy; Blood; CCR; Cancer Detection; Cancer Model; Carcinogenesis Mechanism; Cells; Certification; Chemical Shift Imaging; Chemicals; Chills; Chromatography; Clinic; Clinical; Data Collection; Detection; Deuterium; Development; Dose; Electron Spin Resonance Spectroscopy; Glucose; Glycolysis; Goals; Head; Human; Image; Immune system; In Situ; Infusion procedures; Investigation; Ions; Isotope Labeling; Isotopes; Label; Lead; Lipids; Magic; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Malignant neoplasm of brain; Malignant neoplasm of prostate; Mammals; Mass Spectrum Analysis; Metabolic; Metabolism; Methodology; Methods; Monitor; NMR Spectroscopy; Nuclear; Operative Surgical Procedures; Oxygen; Pathway interactions; Patients; Physiologic pulse; Plants; Positron-Emission Tomography; Preparation; Procedures; Production; Publications; Pyruvate; Pyruvic Acid; Renal Cell Carcinoma; Renal carcinoma; Research; Resolution; Resources; Sampling; Scanning; Specimen; Sterility; Succinate Dehydrogenase; TOCSY; Therapeutic Intervention; Time; Tissues; Toxic effect; Tracer; Tumor-Derived; United States National Institutes of Health; Urine; X-Ray Computed Tomography; cancer biomarkers; cancer diagnosis; cancer imaging; cancer prevention; cancer recurrence; cancer therapy; clinical center; design; experimental study; fluorodeoxyglucose; glucose uptake; imaging capabilities; imaging study; in vivo; inhibitor/antagonist; instrument; isotope incorporation; mass spectrometer; metabolic imaging; metabolomics; method development; molecular imaging; multimodality; new therapeutic target; novel; novel strategies; novel therapeutics; pre-clinical; small molecule; solid state; spectroscopic imaging; stable isotope; tumor; tumor metabolism; ultra high resolution

This facility is focused on the application of targeted, ultra-high resolution stable isotope-resolved metabolomics as well as critical careful sample extraction and preparation methodologies designed to obtain as much information as possible from clinical specimens. The initial mass spectrometer is a Thermo Orbitrap Fusion Lumos capable of 1,000,000 resolving power. This spectrometer is equipped with an Advion Nanomate direct infusion electrospray module as well as an Ion Chromatography module, which together allow for resolution of a wide range of intermediary metabolites. The ultra-high resolution capability of this spectrometer allows for resolution of metabolites that are multiply-labeled with different isotopes (e.g. 13C, 15N, 2H) in a single specimen. We are currently developing a workflow tailored for targeted quantitation of approximately 120 cellular metabolites with the capability for multiple heavy isotope labels in a single experiment. The facility is also equipped with a powerful 700MHz NMR magnet with with a triple inverse resonance cryoprobe and high-capacity chilled autosampler available for targeted and untargeted, isotope-resolved studies of polar and non-polar metabolite and lipid extracts. In situ solid-state tissue metabolite analysis is also available via magic angle spinning (MAS-NMR) Examples include the determination of the global fate of intracellular 13C tracer molecule metabolism in tumor-derived cells such as succinate dehydrogenase-deficient renal cell carcinoma (Saxena et al., 2016), and global assessment of lipid isotope incorporation using 1D and 2D HSQC pulse sequences (Crooks et al., 2018; Lin et al., 2021). In addition, positional determination of metabolite isotope labeling can be determined using 1D and 2D NMR pulse sequences such as 1H-1H TOCSY (Kishimoto et al., 2019). Data collection and method development for NMR analysis of metabolite extracts from clinical tissue specimens obtained during surgical and biopsy procedures in the NIH Clinical Center are currently underway analyses performed using this instrument have already resulted in numerous CCR publications. Pre-clinical metabolic imaging resources are available for dynamic small animal metabolic imaging via Dynamic Nuclear Polarization (hyperpolarization) as well as non-hyperpolarized deuterium and 13C metabolic imaging using image deconvolution algorithms developed at CCR. Additionally, pre-clinical PET-CT imaging capability will be available in 2021. Recent research highlights include quantitative assessment of the relationship between tumor glycolysis and oxygenation status by multimodal, sequential application of electron paramagnetic resonance imaging of tumor oxygen levels, hyperpolarized MRI to assess tumor lactate production rates, and 18FDG-PET imaging to assess tumor glucose uptake (Yamamoto et al., 2020). In addition, a tensor decomposition post-processing algorithm developed at CCR was applied to MRI chemical shift imaging experiments in order to visualize the metabolism of 13C-glucose in tumors in vivo and assess tumor glycolytic propensities without the need for tracer hyperpolarization (Brender et al., 2019). Finally, the activity of a novel therapeutic LDH inhibitor molecule developed at CCR was assessed in vivo using hyperpolarized 13C-pyruvate infusion before and after treatment, revealing potent on-target efficacy and a rapid rewiring of tumor metabolism following therapy (Oshima et al., 2020). Clinical 13C-hyperpolarized metabolic imaging with the investigational agent 1-13C-pyruvic acid will be conducted at NCI in patients with renal, prostate, and brain cancers using a SPINLab clinical polarizer and specialized 13C MRI scanner located in the Molecular Imaging Clinic. Clinical head, abdominal, and endorectal coils are currently under development. Construction of a sterile filling room in the Molecular Imaging Clinic has been completed to facilitate preparation of clinical 13C1-pyruvate doses required for hyperpolarized imaging studies, and the sterile filling facility is currently undergoing certification so that an IND can be cross-filed and scanning of patients can begin.

该设施的重点是应用目标超高分辨率稳定的同位素分辨代谢组学以及旨在从临床标本中获取尽可能多信息的批准仔细的样品提取和制备方法。初始质谱仪是一种能够具有1,000,000分辨率的热轨道融合液体。该光谱仪配备了纳米酸盐直接输注电喷雾模块以及离子色谱模块，该模块共同解决了广泛的中介代谢物。该光谱仪的超高分辨率能力可以分辨出单个样品中用不同同位素（例如13c，15n，2h）多重标记的代谢产物。我们目前正在开发一个针对大约120个细胞代谢产物的靶向定量定量的工作流，具有单个实验中多个重型同位素标签的能力。该设施还配备了功能强大的700MHz NMR磁铁，并具有三重逆共振冷冻探针和高容量的冷藏自动采样器，可用于靶向和无靶标，同位素分辨的研究，对极性和非极性代谢产物和非偏压代谢物和脂质提取物。还可以通过魔法角度旋转（MAS-NMR）实例获得原位固态组织代谢产物分析，包括确定肿瘤衍生细胞中细胞内13C示踪分子代谢的全球命运，例如苯有利酶脱氢酶缺乏的肾脏肾细胞瘤，并使用ISOT ISSQ ISOT ISOT和Global Iset and。序列（Crooks等，2018； Lin等，2021）。此外，可以使用1D和2D NMR脉冲序列（例如1H-1H Tocsy）确定代谢物同位素标记的位置测定（Kishimoto等，2019）。目前，使用该仪器进行了许多CCR出版物，目前正在进行大量的CCR出版物，目前正在进行使用该仪器进行分析的NIH临床中心手术和活检程序中，用于在手术和活检过程中获得的NMR分析的数据收集和方法开发。临床前代谢成像资源可用于通过动态核极化（超极化）以及非高质氘化和13C代谢成像，使用在CCR上开发的图像反趋化算法，可用于动态小动物代谢成像。 Additionally, pre-clinical PET-CT imaging capability will be available in 2021. Recent research highlights include quantitative assessment of the relationship between tumor glycolysis and oxygenation status by multimodal, sequential application of electron paramagnetic resonance imaging of tumor oxygen levels, hyperpolarized MRI to assess tumor lactate production rates, and 18FDG-PET imaging to assess tumor glucose uptake （Yamamoto等，2020）。此外，在CCR上开发的张量分解后处理算法被应用于MRI化学移位成像实验，以可视化体内肿瘤中13C-葡萄糖的代谢，并评估肿瘤糖酵解的预测，而无需进行截止性超极化（Brender Experalization）（Brender Explalization）（Brender等。，2019年）。最后，使用高极化13c-丙酮酸在治疗前后，在体内评估了在CCR上开发的新型治疗性LDH抑制剂分子的活性，揭示了有效的促靶性功效和治疗后肿瘤代谢的快速重新布线（Oshima等人，2020年）。使用Spinlab临床偏光层和位于分子成像诊所的肾脏，前列腺癌和脑部癌的患者，将在NCI对NCI进行临床13C-磨化的代谢成像1-13C-丙酸1-13C-丙酸。目前正在开发临床头，腹部和内直肠线圈。已经完成了分子成像诊所中无菌填充室的建造，以促进高极化成像研究所需的临床13C1-丙酮酸剂量的准备，并且无菌填充设施目前正在认证中，以便可以交叉进行IND和患者的扫描。