C-NMR: a simple yet comprehensive method for analysis of intermediary metabolism

Jeffrey, F. M H; Rajagopal, Arun; Malloy, Craig R.; Sherry, A. Dean

doi:10.1016/0968-0004(91)90004-f

Cited by 106 publications

(90 citation statements)

References 26 publications

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“…Lactate labeling results from the reaction catalyzed by the enzyme lactate dehydrogenase (LDH) and alanine labeling from a transamination reaction catalyzed by alanine transaminase. 13 C-labeled carbon dioxide is formed by the irreversible decarboxylation of [1][2][3][4][5][6][7][8][9][10][11][12][13] C]pyruvate in the reaction catalyzed by the mitochondrial enzyme, pyruvate dehydrogenase (20). This 13 CO 2 rapidly In many cases, higher unpublished levels of polarization have been achieved with these molecules, and in some cases a gadolinium chelate has also been added to improve polarization (47).…”

Section: [1-13 C]pyruvatementioning

confidence: 99%

“…Intravenous injection of [1-13 C]pyruvate can result in the appearance of [1-13 C]lactate, [1][2][3][4][5][6][7][8][9][10][11][12][13] C]alanine, and 13 CO 2 resonances, depending on the tissue probed and its metabolic state. Lactate labeling results from the reaction catalyzed by the enzyme lactate dehydrogenase (LDH) and alanine labeling from a transamination reaction catalyzed by alanine transaminase.…”

Section: [1-13 C]pyruvatementioning

confidence: 99%

“…To date, [1][2][3][4][5][6][7][8][9][10][11][12][13] C]pyruvate has been the most commonly used metabolite for DNP. Intravenous injection of [1-13 C]pyruvate can result in the appearance of [1-13 C]lactate, [1][2][3][4][5][6][7][8][9][10][11][12][13] C]alanine, and 13 CO 2 resonances, depending on the tissue probed and its metabolic state.…”

Section: [1-13 C]pyruvatementioning

confidence: 99%

“…In contrast, 13 C-MRS measurements of cellular metabolism in systems incubated with 13 C-labeled cell substrates give a dynamic, and therefore potentially more useful, measurement of tissue metabolism (6)(7)(8). For example, dynamic measurements of 13 C-labeled glucose incorporation into muscle glycogen have been used to dissect the relative importance of glucose transport and hexokinase and glycogen synthase activity in controlling muscle glycogen synthesis (9).…”

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Tumor imaging using hyperpolarized ¹³C magnetic resonance spectroscopy

Brindle¹,

Bohndiek²,

Gallagher³

et al. 2011

Magnetic Resonance in Med

236

355

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Dynamic nuclear polarization is an emerging technique for increasing the sensitivity of magnetic resonance imaging and spectroscopy, particularly for low-g nuclei. The technique has been applied recently to a number of 13 C-labeled cell metabolites in biological systems: the increase in signal-to-noise allows the spatial distribution of an injected molecule to be imaged as well as its metabolic product or products. This review highlights the most significant molecules investigated to date in preclinical cancer models, either in terms of their demonstrated metabolism in vivo or the biological processes that they can probe. Key words: hyperpolarized carbon-13; dynamic nuclear polarization; imaging; metabolism; tumor; pyruvateThe application of magnetic resonance in medicine has been dominated by imaging of tissue anatomy. However, one of the great strengths of magnetic resonance (MR) is spectroscopy, which allows imaging of tissue biochemistry; this was recognized in the earliest applications of MR to intact biological systems (1). Although magnetic resonance imaging (MRI) has become a routine clinical tool, the use of magnetic resonance spectroscopy (MRS) in patients has lagged far behind; this has been primarily due to a lack of sensitivity, which leads to long measurement times and poor image resolution (2-5). In spite of this, 1 H-MRS measurements of cellular metabolites in a variety of tumor types have been shown to provide a sensitive means to diagnose disease and detect response to treatment (2-4). However, these measurements generally give a static picture of cellular metabolism.In contrast, 13 C-MRS measurements of cellular metabolism in systems incubated with 13 C-labeled cell substrates give a dynamic, and therefore potentially more useful, measurement of tissue metabolism (6-8). For example, dynamic measurements of 13 C-labeled glucose incorporation into muscle glycogen have been used to dissect the relative importance of glucose transport and hexokinase and glycogen synthase activity in controlling muscle glycogen synthesis (9). However, the problem of low sensitivity is even more acute in the case of 13 C-MRS. Dynamic nuclear polarization (DNP or hyperpolarization) of 13 C-labeled cell substrates enhances their sensitivity to detection by >10 4 -fold (10,11). Subsequent spectroscopic imaging of their metabolism following intravenous injection offers a solution to the problem of low sensitivity and has the potential to make dynamic metabolic imaging using MRS a routine clinical application.There have been a number of recent review articles and book chapters on this subject (12-17), and therefore, the purpose of this brief review is to: highlight those DNP substrates that have shown the greatest promise for oncological applications in vivo; summarize the biochemical mechanisms responsible for label transfer from pyruvate to other metabolites in tumors; and finally provide an overview of the main challenges in label detection and imaging and how these may be addressed. PROMISING SUBSTRATES FOR HYPERPOLA...

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Section: [1-13 C]pyruvatementioning

confidence: 99%

Section: [1-13 C]pyruvatementioning

confidence: 99%

Section: [1-13 C]pyruvatementioning

confidence: 99%

mentioning

confidence: 99%

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Tumor imaging using hyperpolarized ¹³C magnetic resonance spectroscopy

Brindle¹,

Bohndiek²,

Gallagher³

et al. 2011

Magnetic Resonance in Med

236

355

View full text Add to dashboard Cite

show abstract

“…The pO 2 difference and measure of coronary flow were used to evaluate MVO 2 (30). Hearts were then freeze-clamped, ground to a fine powder, extracted, and finally dissolved in D 2 O for 13 C and 1 H analysis (31,32). Total substrate oxidation or citric acid cycle flux was determined by using the reported relationship (33,34), 13 C 4 ]acetoacetate under condition ii).…”

Section: Determination Of Lpl Activity In Tissues and Postheparin Plamentioning

confidence: 99%

Inhibition of cardiac lipoprotein utilization by transgenic overexpression of Angptl4 in the heart

Burgess

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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To investigate the role of Angptl4, an inhibitor of lipoprotein lipase that is induced by >3-fold in the heart after rosiglitazone treatment, we generated transgenic mice that overexpress Angptl4 in the heart (MHC-Angptl4). We show that MHC-Angptl4 mice exhibit cardiacrestricted expression of the transgene and inhibition of cardiac lipoprotein lipase (LPL) activity. However, LPL activities in other tissues or that released into plasma by heparin are not affected. In addition, MHC-Angptl4 mice also exhibit hypertriglyceridemia after 6 h of fasting. We use echocardiography to show that MHC-Angptl4 mice develop left-ventricular dysfunction. Comparison of the metabolic profiles of isolated working hearts demonstrates that cardiac impairment in MHC-Angptl4 mice is positively associated with decreased triglyceride (TG) utilization. When bred to transgenic mice that overexpress acyl-CoA synthetase in the heart, a strain that exhibits elevated cardiac TG accumulation, cardiac TG content in double transgenic mice is reversed to that of wild-type mice. Taken together, our data support the hypothesis that induction of Angptl4 in the heart inhibits lipoprotein-derived fatty acid delivery. This mouse model will be useful to elucidate the role of reduced fatty acid supply in the pathogenesis of heart failure and related disorders.lipase ͉ nuclear receptor ͉ cardiomyopathy

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Metabolic flux analysis of hybridoma cells in different culture media using mass balances

Bonarius

Hatzimanikatis

Meesters

et al. 1996

Biotechnol. Bioeng.

242

174

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The estimation of the intracellular fluxes of mammalian cells using only the mass balances of the relevant metabolites is not possible because the set of linear equations defined by these mass balances is underdetermined. Either additional experimental flux data or additional theoretical constraints are required to find one unique flux distribution out of the solution space that is bound by the mass balances. Here, a method is developed using the latter approach. The uptake and production rates of amino acids, glucose, lactate, O2, CO2, NH4, MAB, and the intracellular amino acid pools have been determined for two different steady‐states. The cellular composition {total protein and protein composition, total lipids and fatty acid distribution, total carbohydrates, DNA and RNA} has been measured to calculate the requirements for biosynthesis. It is shown to be essential to determine the uptake/production rates of ammonia and either carbon dioxide or oxygen. In mammalian cells these are cometabolites of cyclic metabolic pathways. The flux distribution that is found using the Euclidean minimum norm as the additional theoretical constraint and taking either the CO2 or the NAD(P)H mass balance into account is shown to be in agreement with the measured O2 and CO2 metabolic rates. The metabolic fluxes in hybridoma cells in continuous culture at a specific growth rate of 0.83 day−1 are estimated for a medium with (optimal medium) and without (suboptimal medium) Primatone RL, an enzymatic hydrolysate of animal tissue that causes a more than twofold increase in cell density. It is concluded that The majority of the consumed glucose (>90%) is channeled through the pentose‐phosphate pathway in rapidly proliferating cells. Pyruvate oxidation and tricarboxylic acid (TCA) cycle activity are relatively low, i.e., 8% of the glucose uptake in suboptimal and 14% in optimal medium, respectively. Under both conditions, only a small fraction of pyruvate is further oxidized to CO2. The flux from glutamate to α‐ketoglutarate (catalyzed by glutamate dehydrogenase) is almost zero in medium with and even slightly reversed in medium without Primatone RL. Almost all glutamate enters the TCA cycle due to the action of transaminases. Transhydrogenation plays a significant role in hybridoma cells under our experimental conditions. NADPH is produced at relatively high rates (11 × 10−12 to 13 × 10−12 mol · cell−1 · day−1) compared to other fluxes in both culture media. © 1996 John Wiley & Sons, Inc.

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C-NMR: a simple yet comprehensive method for analysis of intermediary metabolism

Cited by 106 publications

References 26 publications

Tumor imaging using hyperpolarized ¹³C magnetic resonance spectroscopy

Tumor imaging using hyperpolarized ¹³C magnetic resonance spectroscopy

Inhibition of cardiac lipoprotein utilization by transgenic overexpression of Angptl4 in the heart

Metabolic flux analysis of hybridoma cells in different culture media using mass balances

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C-NMR: a simple yet comprehensive method for analysis of intermediary metabolism

Cited by 106 publications

References 26 publications

Tumor imaging using hyperpolarized 13C magnetic resonance spectroscopy

Tumor imaging using hyperpolarized 13C magnetic resonance spectroscopy

Inhibition of cardiac lipoprotein utilization by transgenic overexpression of Angptl4 in the heart

Metabolic flux analysis of hybridoma cells in different culture media using mass balances

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Tumor imaging using hyperpolarized ¹³C magnetic resonance spectroscopy

Tumor imaging using hyperpolarized ¹³C magnetic resonance spectroscopy