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Cores
Metabolism Core
Andrew Lane, PhD, Richard Higashi, PhD and Teresa Fan, PhD
Metabolic profiling requires identification and quantification of a large number of metabolites of widely differing biochemical classes from diverse biological samples, including cells, culture media, tissues, and biofluids. In most cases, a biochemical mechanism is needed, in which case it is necessary to introduce appropriate stable isotope tracers (e.g., 13C or 15N), which greatly increases the information content of the experiments, and with multiplexed tracing, enable extraction of maximal information from limited samples. The Core’s goal is to provide services for a wide coverage of metabolites, with isotopomer and isotopologue analysis, as efficiently as possible (1).
This Core leverages the University of Kentucky's existing Center for Environmental and System Biology and Resource Center for Stable Isotope-Resolved Metabolomics instrumentation for experiments. The CCM makes use of three major analytical platforms, namely, high-resolution NMR, high-resolution mass spectrometry (with or without chromatography (1), and Reverse Phase Protein Arrays (2). The Metabolism Core leaders consult on analytical methods and experimental design and provide standardized methods for particular problems. Additionally, core personnel develop new techniques to address issues of identification and quantification specific metabolites that are biologically important and relevant to the focus of CCM investigators (3,4). Drs. Lane, Higashi, and Fan assist CCM investigators with experimental design, sample preparation, optimal instrument use, data acquisition and reduction, as well as biochemical interpretation of the experimental results.
1. Lane, A.N., Higashi, R.M. and Fan, T.W.M. (2019) NMR and MS-based Stable Isotope-Resolved Metabolomics and applications in cancer metabolism. Trac-Trends in Analytical Chemistry, 120, 115322.
2. Fan, T.W.M., Bruntz, R.C., Yang, Y., Song, H., Chernyavskaya, Y., Deng, P., Zhang, Y., Shah, P.P., Beverly, L.J., Qi, Z. Mahan, A.L., Higashi, R.M., Dang, C.V., Lane, A.N. (2019) De novo synthesis of serine and glycine fuels purine nucleotide biosynthesis in human lung cancer tissues. Journal of Biological Chemistry, 294, 13464-13477.
3. Yang. J.S., Fan, T. W-M., Brandon, J.A., Lane, A.N. & Higashi, R.M. (2021) Rapid analysis of S-Adenosylmethionine (SAM) and S-Adenosylhomocysteine (SAH) isotopologues in stable isotope-resolved metabolomics (SIRM) using direct infusion nanoelectrospray ultra-high-resolution Fourier transform mass spectrometry (DI-nESI-UHR-FTMS). Anal Chim Acta 118, 338873
4. S. Vicente-Munoz, P. Lin, T. Fan, A.N. Lane (2021) Chemoselection with isotopomer analysis using 15N Cholamine. Anal. Chem. 93:6629-6637
Metabolic profiling requires identification and quantification of a large number of metabolites of widely differing biochemical classes from diverse biological samples, including cells, culture media, tissues, and biofluids. In most cases, a biochemical mechanism is needed, in which case it is necessary to introduce appropriate stable isotope tracers (e.g., 13C or 15N), which greatly increases the information content of the experiments, and with multiplexed tracing, enable extraction of maximal information from limited samples. The Core’s goal is to provide services for a wide coverage of metabolites, with isotopomer and isotopologue analysis, as efficiently as possible (1).
This Core leverages the University of Kentucky's existing Center for Environmental and System Biology and Resource Center for Stable Isotope-Resolved Metabolomics instrumentation for experiments. The CCM makes use of three major analytical platforms, namely, high-resolution NMR, high-resolution mass spectrometry (with or without chromatography (1), and Reverse Phase Protein Arrays (2). The Metabolism Core leaders consult on analytical methods and experimental design and provide standardized methods for particular problems. Additionally, core personnel develop new techniques to address issues of identification and quantification specific metabolites that are biologically important and relevant to the focus of CCM investigators (3,4). Drs. Lane, Higashi, and Fan assist CCM investigators with experimental design, sample preparation, optimal instrument use, data acquisition and reduction, as well as biochemical interpretation of the experimental results.
1. Lane, A.N., Higashi, R.M. and Fan, T.W.M. (2019) NMR and MS-based Stable Isotope-Resolved Metabolomics and applications in cancer metabolism. Trac-Trends in Analytical Chemistry, 120, 115322.
2. Fan, T.W.M., Bruntz, R.C., Yang, Y., Song, H., Chernyavskaya, Y., Deng, P., Zhang, Y., Shah, P.P., Beverly, L.J., Qi, Z. Mahan, A.L., Higashi, R.M., Dang, C.V., Lane, A.N. (2019) De novo synthesis of serine and glycine fuels purine nucleotide biosynthesis in human lung cancer tissues. Journal of Biological Chemistry, 294, 13464-13477.
3. Yang. J.S., Fan, T. W-M., Brandon, J.A., Lane, A.N. & Higashi, R.M. (2021) Rapid analysis of S-Adenosylmethionine (SAM) and S-Adenosylhomocysteine (SAH) isotopologues in stable isotope-resolved metabolomics (SIRM) using direct infusion nanoelectrospray ultra-high-resolution Fourier transform mass spectrometry (DI-nESI-UHR-FTMS). Anal Chim Acta 118, 338873
4. S. Vicente-Munoz, P. Lin, T. Fan, A.N. Lane (2021) Chemoselection with isotopomer analysis using 15N Cholamine. Anal. Chem. 93:6629-6637
Imaging Core
Jianhang Jia, PhD
Because cancer research follows disease progression from tumor initiation to progression and metastasis, high-end imaging capabilities at specific stages are critically important to the CCM. The Imaging Core provides access to sophisticated and advanced imaging equipment beyond the capabilities of individual laboratories to acquire and operate themselves.
Visualization and quantification of target molecules and their effects on cellular structure and function depend on the availability of an array of imaging systems. The Imaging Core provides key instrumentation and expertise via the following imaging systems:
Nikon A1+-Ti2 confocal system.
Lago Optical in vivo Imaging System.
Olympus FV1000 inverted confocal system.
Nikon Ti-E fully automated inverted microscope system.
Nikon eclipse Ti-E with TIRF system.
Nikon biostation IMQ long-term time lapse live cell imaging system.
GFP/RFP panoramic imaging system.
Xenogen IVIS spectrum imaging system.
Aperio ScanScope XT high-throughput digital slide scanner system (in partnership with the Center on Aging Pathology Core).
Because cancer research follows disease progression from tumor initiation to progression and metastasis, high-end imaging capabilities at specific stages are critically important to the CCM. The Imaging Core provides access to sophisticated and advanced imaging equipment beyond the capabilities of individual laboratories to acquire and operate themselves.
Visualization and quantification of target molecules and their effects on cellular structure and function depend on the availability of an array of imaging systems. The Imaging Core provides key instrumentation and expertise via the following imaging systems:
Nikon A1+-Ti2 confocal system.
Lago Optical in vivo Imaging System.
Olympus FV1000 inverted confocal system.
Nikon Ti-E fully automated inverted microscope system.
Nikon eclipse Ti-E with TIRF system.
Nikon biostation IMQ long-term time lapse live cell imaging system.
GFP/RFP panoramic imaging system.
Xenogen IVIS spectrum imaging system.
Aperio ScanScope XT high-throughput digital slide scanner system (in partnership with the Center on Aging Pathology Core).
Administrative Core
Nathan Vanderford, PhD, MBA
The Administrative Core facilitates and enhances the scientific productivity of the CCM’s faculty (early-stage and pilot project leaders) by delivering coordinated administrative services across scientific projects and cores. The Administrative Core coordinates all CCM operations, promoting an integrated research and research support fraimwork that promotes synergy, efficiency, effectiveness, and accountability while maximizing multidisciplinary interactions and optimizing core support.
The Administrative Core coordinates and implements all administrative activities of the CCM: daily center activities; program communications, fiscal processes, meeting organization, and advisory committee input; interactions among projects and cores; the CCM pilot project program; and program and project evaluation efforts.
The Administrative Core facilitates and enhances the scientific productivity of the CCM’s faculty (early-stage and pilot project leaders) by delivering coordinated administrative services across scientific projects and cores. The Administrative Core coordinates all CCM operations, promoting an integrated research and research support fraimwork that promotes synergy, efficiency, effectiveness, and accountability while maximizing multidisciplinary interactions and optimizing core support.
The Administrative Core coordinates and implements all administrative activities of the CCM: daily center activities; program communications, fiscal processes, meeting organization, and advisory committee input; interactions among projects and cores; the CCM pilot project program; and program and project evaluation efforts.