Metabolism of alternative substrates and the bioenergetic status of EMT6 tumor cell spheroids

Wehrle, Janna P.; Ng, Cheng E.; McGovern, Kathy Ann; Aiken, Nanci R.; Shungu, Dikoma C.; Chance, Edwin M.; Glickson, Jerry D.

doi:10.1002/1099-1492(200010)13:6<349::aid-nbm652>3.0.co;2-x

Cited by 25 publications

(30 citation statements)

References 23 publications

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“…3D culture or spheroids can be generated from established cell lines by growing them on an appropriate matrix (such as collagen ± laminin synthetic extracellular matrix, hydrogel scaffolds, among many others (Lee J et al 2008; Ravi et al 2014)), such that they spontaneously form 3D structures that in many cases mimic those found in natural tissue (Bissell et al 2003) (G. Y. Lee et al 2007; Wehrle et al 2000). It is also possible to generate spheroids or whole organoids containing multiple cell types (G.…”

Section: Preclinical Modelsmentioning

confidence: 99%

“…It is also possible to generate spheroids or whole organoids containing multiple cell types (G. Y. Lee et al 2007; Willyard 2015; McCracken et al 2014; Spence et al 2011), which much more closely mimic the natural tissue, while retaining the flexibility for experimental manipulation in much the same way as 2D cultures (Wehrle et al 2000; Pawlik et al 2000). Such systems have been shown to display altered gene expression profiles and drug response compared with the corresponding 2D cell culture systems (Ravi et al 2014).…”

Section: Preclinical Modelsmentioning

confidence: 99%

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Preclinical models for interrogating drug action in human cancers using Stable Isotope Resolved Metabolomics (SIRM)

2016

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Aims In this review we compare the advantages and disadvantages of different model biological systems for determining the metabolic functions of cells in complex environments, how they may change in different disease states, and respond to therapeutic interventions. Background All preclinical drug-testing models have advantages and drawbacks. We compare and contrast established cell, organoid and animal models with ex vivo organ or tissue culture and in vivo human experiments in the context of metabolic readout of drug efficacy. As metabolism reports directly on the biochemical state of cells and tissues, it can be very sensitive to drugs and/or other environmental changes. This is especially so when metabolic activities are probed by stable isotope tracing methods, which can also provide detailed mechanistic information on drug action. We have developed and been applying Stable Isotope-Resolved Metabolomics (SIRM) to examine metabolic reprogramming of human lung cancer cells in monoculture, in mouse xenograft/explant models, and in lung cancer patients in situ (Lane et al. 2011; T. W. Fan et al. 2011; T. W-M. Fan et al. 2012; T. W. Fan et al. 2012; Xie et al. 2014b; Ren et al. 2014a; Sellers et al. 2015b). We are able to determine the influence of the tumor microenvironment using these models. We have now extended the range of models to fresh human tissue slices, similar to those originally described by O. Warburg (Warburg 1923), which retain the native tissue architecture and heterogeneity with a paired benign versus cancer design under defined cell culture conditions. This platform offers an unprecedented human tissue model for preclinical studies on metabolic reprogramming of human cancer cells in their tissue context, and response to drug treatment (Xie et al. 2014a). As the microenvironment of the target human tissue is retained and individual patient's response to drugs is obtained, this platform promises to transcend current limitations of drug selection for clinical trials or treatments. Conclusions and Future Work Development of ex vivo human tissue and animal models with humanized organs including bone marrow and liver show considerable promise for analyzing drug responses that are more relevant to humans. Similarly using stable isotope tracer methods with these improved models in advanced stages of the drug development pipeline, in conjunction with tissue biopsy is expected significantly to reduce the high failure rate of experimental drugs in Phase II and III clinical trials.

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Section: Preclinical Modelsmentioning

confidence: 99%

Section: Preclinical Modelsmentioning

confidence: 99%

Preclinical models for interrogating drug action in human cancers using Stable Isotope Resolved Metabolomics (SIRM)

2016

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“…One study used NMR on murine mammary carcinoma/sarcoma EMT6 tumor cell spheroids to obtain evidence of fatty acid degradation in cancer cells. 13 C-propionate was used to monitor metabolic changes in the spheroids (with glucose and glutamine present) and the radiolabel was incorporated into succinate, malate and glutamate, supporting direct fatty acid degradation (Wehrle et al, 2000).…”

Section: Fatty Acid Degradation By B-oxidationmentioning

confidence: 99%

“…When several carcinomas (HeLa, AS-30D) were cultured in glucose-rich but Chang et al (1991), Mazurek et al (1997) glutamine-depleted medium, tumor growth became significantly diminished, indicating that glutamine oxidation supplies ATP, and intermediates for anabolic pathways Drogat et al, 2007;Donadio et al, 2008). Several other studies have also shown the adaptability of cancer cells to switch to using either glucose or glutamine interchangeably as alternative substrates for oxidative growth (Reitzer et al, 1979;Baggetto, 1992;Wehrle et al, 2000;Rossignol et al, 2004).…”

Section: Glutamate Dehydrogenasementioning

confidence: 99%

“…Propionate was shown to be readily utilized as a substrate by Caco-2 colon cancer cells (Malaisse et al, 1996) and was also readily used by the murine mammary carcinoma/sarcoma EMT6 spheroids as an anapleurotic substrate. In the tumor spheroids, propionate was converted to methylmalonate and succinate even in the presence of normal levels of glucose and glutamine in the media (Wehrle et al, 2000). Hence, although use of propionate by tumors in vivo has not been extensively studied, the above data indicate that tumor cells are highly likely to metabolize propionate.…”

Section: Ketone Body (Kb) Metabolismmentioning

confidence: 99%

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Bioenergetic pathways in tumor mitochondria as targets for cancer therapy and the importance of the ROS-induced apoptotic trigger

Ralph

Rodrı́guez-Enrı́quez

Neužil

et al. 2010

Molecular Aspects of Medicine

143

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Mitochondria are emerging as idealized targets for anti-cancer drugs. One reason for this is that although these organelles are inherent to all cells, drugs are being developed that selectively target the mitochondria of malignant cells without adversely affecting those of normal cells. Such anti-cancer drugs destabilize cancer cell mitochondria and these compounds are referred to as mitocans, classified into several groups according to their mode of action and the location or nature of their specific drug targets. Many mitocans selectively interfere with the bioenergetic functions of cancer cell mitochondria, causing major disruptions often associated with ensuing overloads in ROS production leading to the induction of the intrinsic apoptotic pathway. This in-depth review describes the bases for the bioenergetic differences found between normal and cancer cell mitochondria, focussing on those essential changes occurring during malignancy that clinically may provide the most effective targets for mitocan development. A common theme emerging is that mitochondrially mediated ROS activation as a trigger for apoptosis offers a powerful basis for cancer therapy. Continued research in this area is likely to identify increasing numbers of novel agents that should prove highly effective against a variety of cancers with preferential toxicity towards malignant tissue, circumventing tumor resistance to the other more established therapeutic anti-cancer approaches.

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Real‐time detection of ¹³C NMR labeling kinetics in perfused EMT6 mouse mammary tumor cells and βHC9 mouse insulinomas

Mancuso

Beardsley

Wehrli

et al. 2004

Biotech & Bioengineering

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A method was developed for obtaining high signal-to-noise 13C NMR spectra of intracellular compounds in metabolically active cultured cells. The method allows TCA cycle labeling kinetics to be determined in real time without significant oxygen transport limitations. Cells were immobilized on the surface of nonporous microcarriers that were either uncoated or coated with polypeptides and used in a 12-cm3 packed bed. The methods were tested with two EMT6 mouse mammary tumor cell lines, one strongly adherent and the other moderately adherent, and a weakly adherent mouse insulinoma line (betaHC9). For both EMT6 lines, NTP and oxygen consumption measurements indicated that the number of cells in the spectrometer ranged from 6 x 10(8) to 1 x 10(9). During infusion of [1-13C]glucose, labeling in C-4 glutamate (indicative of flux into the first half of the TCA cycle) could be detected with 15-min resolution. However, labeling for C-3 and C-2 glutamate (indicative of complete TCA cycle activity) was fivefold lower and difficult to quantify. To increase TCA cycle labeling, cells were infused with medium containing [1,6-13C2]glucose. A 2.5-fold increase was observed in C-4 glutamate labeling and C-3 and C-2 glutamate labeling could be monitored with 30-min resolution. Citrate synthase activity was indirectly detected in real time, as [3,4-13C2]glutamate was formed from [2-13C]oxaloacetate and [2-13C]acetate (of acetyl-CoA). Cell mass levels observed with betaHC9 cells were somewhat lower. However, the 13C S/N was sufficient to allow real-time monitoring of the response of intracellular metabolite labeling to a step change in glucose and a combined glutamine/serum pulse.

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Metabolism of alternative substrates and the bioenergetic status of EMT6 tumor cell spheroids

Cited by 25 publications

References 23 publications

Preclinical models for interrogating drug action in human cancers using Stable Isotope Resolved Metabolomics (SIRM)

Preclinical models for interrogating drug action in human cancers using Stable Isotope Resolved Metabolomics (SIRM)

Bioenergetic pathways in tumor mitochondria as targets for cancer therapy and the importance of the ROS-induced apoptotic trigger

Real‐time detection of ¹³C NMR labeling kinetics in perfused EMT6 mouse mammary tumor cells and βHC9 mouse insulinomas

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Metabolism of alternative substrates and the bioenergetic status of EMT6 tumor cell spheroids

Cited by 25 publications

References 23 publications

Preclinical models for interrogating drug action in human cancers using Stable Isotope Resolved Metabolomics (SIRM)

Preclinical models for interrogating drug action in human cancers using Stable Isotope Resolved Metabolomics (SIRM)

Bioenergetic pathways in tumor mitochondria as targets for cancer therapy and the importance of the ROS-induced apoptotic trigger

Real‐time detection of 13C NMR labeling kinetics in perfused EMT6 mouse mammary tumor cells and βHC9 mouse insulinomas

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Product

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About

Real‐time detection of ¹³C NMR labeling kinetics in perfused EMT6 mouse mammary tumor cells and βHC9 mouse insulinomas