1. Academic Validation
  2. 13C metabolic flux analysis clarifies distinct metabolic phenotypes of cancer cell spheroid mimicking tumor hypoxia

13C metabolic flux analysis clarifies distinct metabolic phenotypes of cancer cell spheroid mimicking tumor hypoxia

  • Metab Eng. 2022 Sep;73:192-200. doi: 10.1016/j.ymben.2022.07.008.
Shingo Noguchi 1 Kenichi Wakita 2 Fumio Matsuda 3 Hiroshi Shimizu 4
Affiliations

Affiliations

  • 1 Translational Science Department I, Daiichi Sankyo Co., Ltd., 1-2-58, Hiromachi, Shinagawa-ku, Tokyo, 140-8710, Japan; Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5, Yamadaoka, Suita, Osaka, 565-0871, Japan. Electronic address: shingo.noguchi.ni@daiichisankyo.co.jp.
  • 2 Translational Science Department I, Daiichi Sankyo Co., Ltd., 1-2-58, Hiromachi, Shinagawa-ku, Tokyo, 140-8710, Japan. Electronic address: wakita.kenichi.e2@daiichisankyo.co.jp.
  • 3 Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5, Yamadaoka, Suita, Osaka, 565-0871, Japan. Electronic address: fmatsuda@ist.osaka-u.ac.jp.
  • 4 Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5, Yamadaoka, Suita, Osaka, 565-0871, Japan. Electronic address: shimizu@ist.osaka-u.ac.jp.
Abstract

Cancer cells adapt their intracellular energy metabolism to the oxygen-deprived tumor microenvironment (TME) to ensure tumor progression. This adaptive mechanism has focused attention on the metabolic phenotypes of tumor cells under hypoxic TME for developing novel Cancer therapies. Although widely used monolayer (2D) culture does not fully reflect in vivo hypoxic TME, spheroid (3D) culture can produce a milieu similar to the TME in vivo. However, how different metabolic phenotypes are expressed in 3D cultures mimicking tumor hypoxia compared with 2D cultures under hypoxia remains unclear. To address this issue, we investigated the metabolic phenotypes of 2D- and 3D-cultured Cancer cells by 13C-metabolic flux analysis (13C-MFA). Principal component analysis of 13C mass isotopomer distributions clearly demonstrated distinct metabolic phenotypes of 3D-cultured cells. 13C-MFA clarified that 3D culture significantly upregulated pyruvate carboxylase flux in line with the pyruvate carboxylase protein expression level. On the Other hand, 3D culture downregulated glutaminolytic flux. Consistent with our findings, 3D-cultured cells are more resistant to a Glutaminase Inhibitor than 2D-cultured cells. This study suggests the importance of considering the metabolic characteristics of the particular in vitro model used for research on Cancer metabolism.

Keywords

(13)C-metabolic flux analysis; 3D culture; Cancer metabolism; Hypoxic tumor microenvironment; Spheroid.

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