Metabolomics Core

Division Of Basic Sciences - Nci

Investigators

Linked publications & trials

Abstract

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.