Monitoring Early Glycolytic Flux Alterations Following Radiotherapy in Cancer and Immune Cells: Hyperpolarized Carbon-13 Magnetic Resonance Imaging Study

Ying-Chieh Lai^{1

2}, Ching-Yi Hsieh^{2

3

4}, Kuan-Ying Lu^{1

2}, Cheng-Hsuan Sung^{3

4}, Hung-Yao Ho^{2

5

6

7}, Mei-Ling Cheng^{2

5

6

8}, Albert P Chen⁹, Shu-Hang Ng^{1

4}, Fang-Hsin Chen^{4

10

11}, Gigin Lin^{1

2

3

4}

Affiliations

¹ Department of Medical Imaging and Intervention, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan.
² Clinical Metabolomics Core Laboratory, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan.
³ Medical Imaging Research Center, Institute for Radiological Research, Chang Gung University, Taoyuan 333, Taiwan.
⁴ Department of Medical Imaging and Radiological Sciences, Chang Gung University, Taoyuan 333, Taiwan.
⁵ Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
⁶ Metabolomics Core Laboratory, Healthy Aging Research Center, Chang Gung University, Taoyuan 333, Taiwan.
⁷ Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
⁸ Department of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
⁹ GE Healthcare, Toronto, ON M5V 3Y3, Canada.
¹⁰ Radiation Biology Research Center, Institute for Radiological Research, Chang Gung University, Taoyuan 333, Taiwan.
¹¹ Department of Radiation Oncology, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan.

PMID: 34436459
PMCID: PMC8398834
DOI: 10.3390/metabo11080518

Monitoring Early Glycolytic Flux Alterations Following Radiotherapy in Cancer and Immune Cells: Hyperpolarized Carbon-13 Magnetic Resonance Imaging Study

Ying-Chieh Lai et al. Metabolites. 2021.

. 2021 Aug 6;11(8):518.

doi: 10.3390/metabo11080518.

Authors

Affiliations

¹ Department of Medical Imaging and Intervention, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan.
² Clinical Metabolomics Core Laboratory, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan.
³ Medical Imaging Research Center, Institute for Radiological Research, Chang Gung University, Taoyuan 333, Taiwan.
⁴ Department of Medical Imaging and Radiological Sciences, Chang Gung University, Taoyuan 333, Taiwan.
⁵ Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
⁶ Metabolomics Core Laboratory, Healthy Aging Research Center, Chang Gung University, Taoyuan 333, Taiwan.
⁷ Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
⁸ Department of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 333, Taiwan.
⁹ GE Healthcare, Toronto, ON M5V 3Y3, Canada.
¹⁰ Radiation Biology Research Center, Institute for Radiological Research, Chang Gung University, Taoyuan 333, Taiwan.
¹¹ Department of Radiation Oncology, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, Taiwan.

PMID: 34436459
PMCID: PMC8398834
DOI: 10.3390/metabo11080518

Abstract

Alterations in metabolism following radiotherapy affect therapeutic efficacy, although the mechanism underlying such alterations is unclear. A new imaging technique-named dynamic nuclear polarization (DNP) carbon-13 magnetic resonance imaging (MRI)-probes the glycolytic flux in a real-time, dynamic manner. The [1-¹³C]pyruvate is transported by the monocarboxylate transporter (MCT) into cells and converted into [1-¹³C]lactate by lactate dehydrogenase (LDH). To capture the early glycolytic alterations in the irradiated cancer and immune cells, we designed a preliminary DNP ¹³C-MRI study by using hyperpolarized [1-¹³C]pyruvate to study human FaDu squamous carcinoma cells, HMC3 microglial cells, and THP-1 monocytes before and after irradiation. The pyruvate-to-lactate conversion rate (k_PL [Pyr.]) calculated by kinetic modeling was used to evaluate the metabolic alterations. Western blotting was performed to assess the expressions of LDHA, LDHB, MCT1, and MCT4 proteins. Following irradiation, the pyruvate-to-lactate conversion rates on DNP ¹³C-MRI were significantly decreased in the FaDu and the HMC3 cells but increased in the THP-1 cells. Western blot analysis confirmed the similar trends in LDHA and LDHB expression levels. In conclusion, DNP ¹³C-MRI non-invasively captured the different glycolytic alterations among cancer and immune systems in response to irradiation, implying its potential for clinical use in the future.

Keywords: cancer metabolism; dynamic nuclear polarization; glycolysis; immune system; magnetic resonance imaging; radiation.

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Conflict of interest statement

The authors declare that there is no conflicts of interest.

Figures

**Figure 1**
Diagram of the metabolic fates of [1-¹³C]pyruvate that were detected by hyperpolarized ¹³C-MR spectroscopy in this study. Note—MCT, monocarboxylate transporter; LDH, lactate dehydrogenase; ALT, alanine transaminase.

**Figure 2**
A schematic plot of the delivery timeline of hyperpolarized [1-¹³C]pyruvate. Three stages described the delivery steps: (I) dissolution and export of hyperpolarized [1-¹³C]pyruvate from the polarizer; (II) mixing of hyperpolarized [1-¹³C]pyruvate with the cell suspensions; (**III**) start of ¹³C-MR spectroscopy acquisition. Step (II) was defined as “t = 0”.

**Figure 3**
Representative data from an in vitro study. (A) Orange dots represent the data for [1-¹³C]pyruvate (I) and [1-¹³C]lactate (II) obtained from the spectroscopy-based ¹³C-MRI acquisition. Blue dots represent best fit lines calculated by kinetic modeling. (B) Imaging-based ¹³C-MRI acquisition of the investigated metabolites displayed at a temporal resolution of 2 s. Each row represents one metabolite: lactate (chemical shift, 392 Hz), pyruvate–hydrate (267 Hz), alanine (177 Hz), pyruvate (0 Hz), and bicarbonate (−324 Hz). Hyperpolarized ¹³C signals of lactate, pyruvate–hydrate, and pyruvate were visually observed.

**Figure 4**
Comparison of the conversion rates (k_PL [Pyr.]) of hyperpolarized [1-¹³C]pyruvate to [1-¹³C]lactate between the non-irradiated (non-RT) and irradiated (RT) FaDu, HMC3, and THP-1 cells (* p < 0.05; ** p < 0.01). Note—k_PL [Pyr.] unit: nM/s/10⁶ cells.

**Figure 5**
Analysis of the expressions of LDHA and LDHB in the non-irradiated (non-RT) and irradiated (RT) cancer and immune cells. (A) Western blot analyses of LDHA and LDHB in FaDu, HMC3, and THP-1 cells. GAPDH was used as a loading control. (B) Comparison of the expressions of LDHA and LDHB between the non-irradiated and irradiated cells (* p < 0.05; ** p < 0.01). Note—LDH, lactate dehydrogenase; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

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Monitoring Early Glycolytic Flux Alterations Following Radiotherapy in Cancer and Immune Cells: Hyperpolarized Carbon-13 Magnetic Resonance Imaging Study

Affiliations

Monitoring Early Glycolytic Flux Alterations Following Radiotherapy in Cancer and Immune Cells: Hyperpolarized Carbon-13 Magnetic Resonance Imaging Study

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

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