The Center for Clinical Cancer Metabolism provides infrastructure and expertise in the following focused areas of cancer metabolism research:
1) Three-phase sample preparation strategies designed to maximize the information that can be obtained from clinical and pre-clinical tissues and cells.
2) NMR-based multiple isotope-resolved metabolomics of polar and non-polar cell and tissue extracts.
3) Mass spectrometry-based targeted analysis of polar Single- and Multiple Isotope-labeled cell and tissue extracts.
4) Preclinical multinuclear NMR-based metabolic imaging of tumors and other tissues, including Dynamic Nuclear Polarization (DNP) for in vivo metabolic tracer studies.
5) Preclinical EPR mapping and measurement of oxygen concentration in tumors and other tissues.
6) Clinical DNP for hyperpolarized 13C1-pyruvate studies in human subjects at the NIH Clinical Center.
1. Bruker 700Mhz NMR with Triple Inverse HCN cold probe: targeted and untargeted, isotope-resolved studies of polar and non-polar metabolite and lipid extracts. Determination of positional isotopic labeling distribution of tracer molecules in metabolites of interest. High throughput studies made possible with Samplejet 480 position chilled autosampler.
2. Thermo Fusion Lumos Mass Spectrometer with ICS-6000 Ion Chromatograph and Advion Nanomate direct infusion accessory: Instrument is capable of 1,000,000 resolving power at 200 m/z. 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 have developed and implemented an assay tailored for targeted quantitation of approximately 1400 cellular metabolites with the capability for multiple heavy isotope labels in a single experiment.
3. Thermo Exploris 240 Mass Spectrometer with Vanquish Duo Chromatography: This instrument is capable of 240,000 resolving power at 200 m/z. Equipped with a Vanquish Duo chromatograph that is configured for HILIC chromatography. Initial assay workflow is aimed towards quantitative isotope-resolved assessment of the NAD metabolome and other positively charged and zwitterionic metabolite species.
4. SPEX LN2 Tissue Grinder: For grinding of human tissue samples under liquid nitrogen temperatures for metabolite extraction.
5. Lyophilizer (2): For freeze drying frozen aqueous metabolite extracts during sample preparation.
6. Biotek Cytation 5 Plate Reader: Used for measurement of sample protein concentrations and for enzyme assays
7. Vacuum Centrifuge (2): Oil-free vacuum pumps for drying of lipid and extracts from tissues and cells.
8. Data Analysis Workstations (2): Available to facility staff and collaborators for analysis of their NMR and mass spectrometry data.
9. Mestrenova Software, including onsite MNova Database
10. Thermo Compound Discoverer Software
11. Biosafety Cabinet for Tissue Culture
1. Bruker 3T Pre-clinical MRI: This instrument is equipped with multinuclear coils capable of imaging and spectroscopy of 1H, 13C, 15N and 2H. Utilized by many different groups across CCR for both hyperpolarized and non-hyperpolarized metabolic imaging studies.
2. EPR Oximetry Imager: Custom designed and built on site instrument capable of non-invasive imaging of tissue and tumor oxygen concentrations.
3. Spinaligner pre-clinical polarizer: Preclinical hyperpolarizer instrument for performing dynamic nuclear polarization (DNP) experiments to produce metabolic imaging tracers.
4. GE SPINLab Clinical Polarizer: Clinical-grade hyperpolarizer instrument for performing dynamic nuclear polarization (DNP) of 13C-pyruvate doses for injection into humans for metabolic flux imaging.
1. Seahorse XF96e Flux Analyzer: For measurement of respiration and extracellular acidification in tumor cell cultures.
2. YSI 2950 Bioanalyzer: For measurement of glucose, lactate, glutamine and glutamate concentrations in culture media and human plasma samples
The Program is geared toward hypothesis-driven, highly collaborative, and clinically-oriented projects in which investigators and their trainees participate directly with the facility staff in experimental design, sample preparation and extraction, data analysis, informatics, and biological interpretation. Trainees from Collaborators’ labs are expected to come to the facility to prepare and extract their own tissues or cell samples under the guidance of facility staff. Computer workstations are available within the facility for analysis of NMR and mass spectrometry data, with guidance and training provided by the facility staff.
Projects are submitted on a rolling basis and following CCR-wide calls for proposals and reviewed by the CCM Steering Committee for clinical and pre-clinical relevance, and alignment with the current technical capabilities of the facility. Submissions are restricted to CCR labs at this time.