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PD1 blockade alters cell-cycle distribution and affects 3′-deoxy-3′-[18F]fluorothymidine uptake in a mouse CT26 tumor model

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Abstract

Objective

We previously reported that alterations of the tumor microenvironment (TME) by programmed death receptor-1 (PD1) blockade affected tumor glucose metabolism and tumor 2-deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) uptake. In cancer cells, high glycolysis allows cells to sustain rapid proliferation since glycolysis is closely related to the proliferation of cancer cells. Therefore, imaging of cellular proliferation may provide more detail of TME alterations. In this study, we investigated how TME alterations by PD1 blockade affects the uptake of 3′-deoxy-3′-[18F]fluorothymidine ([18F]FLT), which is a 18F-radiolabeled thymidine derivative and is taken up by proliferating cells.

Methods

Mice inoculated with murine colon carcinoma CT26 cells were intraperitoneally administered an anti-PD1 antibody on Day 0, when the tumor volume exceeded 50 mm3, and Day 5. [18F]FLT-PET imaging was performed pre-treatment (Day 0) and post treatment (Day 7). Tumor infiltrating lymphocytes (TILs) were identified by flow cytometry. [18F]FLT accumulation and localization in tumor tissue was evaluated by autoradiography and immunohistochemistry. The cell-cycle distribution of tumors and CT26 cells exposed to cytokines (interleukin-2, interferon [INF]-γ, and tumor necrosis factor [TNF]-α) was analyzed by flow cytometry.

Results

PD1 blockade increased CD8+ and CD4+ T cells in tumor tissue and significantly suppressed tumor proliferation; however, tumor [18F]FLT uptake remained unchanged. Autoradiography and immunohistochemistry showed that [18F]FLT was mainly taken up by cancer cells, but not TILs. Flow cytometric analysis demonstrated that the population of cells in G2/M phase increased after PD1 blockade. Moreover, INF-γ and TNF-α significantly increased cells in G2/M phase in vitro.

Conclusion

PD1 blockade-induced alteration of the TME increased CT26 tumor cells in the G2/M phase, which have high thymidine kinase 1 activity. Therefore, [18F]FLT is taken up by tumor cells even if tumor proliferation is suppressed. This observation may be useful for evaluating the response to immunotherapy.

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Abbreviations

[18F]FDG:

2-Deoxy-2-[18F]fluoro-D-glucose

[18F]FLT:

3′-Deoxy-3′-[18F]fluorothymidine

IFN-γ:

Interferon gamma; IL-2, interleukin-2

PET:

Positron emission tomography

PD1:

Programmed cell death-1

RT:

Room temperature

TME:

Tumor-microenvironment

TIL:

Tumor infiltrating lymphocyte

TK1:

Thymidine kinase 1

TNF-α:

Tumor necrosis factor alpha

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Acknowledgements

This work was supported, in part, by JST CREST Grant Number JPMJCR1902; JSPS KAKENHI Grant Number 19H03593, 20K16813; AMED Grant Number JP21zf0127004; the Photo-excitonix Project in Hokkaido University; Takeda Science Foundation. We would like to thank the staff of the Hokkaido University Hospital Cyclotron Facility for synthesis of [18F]FLT.

Funding

Core Research for Evolutional Science and Technology, JPMJCR1902, Mikako Ogawa, Japan Society for the Promotion of Science, 19H03593, Mikako Ogawa, 20K16813, Motofumi Suzuki, Takeda Science Foundation.

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Authors and Affiliations

  1. Laboratory of Bioanalysis and Molecular Imaging, Graduate School of Pharmaceutical Sciences, Hokkaido University, North 12, West 6, Kita-ku, Sapporo, Hokkaido, 060-0812, Japan

    Motofumi Suzuki, Takuma Matsuda, Kohei Nakajima, Yuta Yokouchi & Mikako Ogawa

  2. Central Institute of Isotope Science, Hokkaido University, Sapporo, Hokkaido, 060-0815, Japan

    Yuji Kuge

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  1. Motofumi Suzuki
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  2. Takuma Matsuda
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  3. Kohei Nakajima
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  4. Yuta Yokouchi
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  6. Mikako Ogawa
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Correspondence to Mikako Ogawa.

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Suzuki, M., Matsuda, T., Nakajima, K. et al. PD1 blockade alters cell-cycle distribution and affects 3′-deoxy-3′-[18F]fluorothymidine uptake in a mouse CT26 tumor model. Ann Nucl Med 36, 931–940 (2022). https://doi.org/10.1007/s12149-022-01782-0

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