Dual-tracer autoradiographic analysis of glucose metabolism and hypoxia in orthotopic and PDX tumor models

Authors

  • Morten Busk Department of Experimental Clinical Oncology, Aarhus University Hospital, Denmark; Danish Centre for Particle Therapy, Aarhus University Hospital, Denmark
  • Martin K Thomsen Department of Biomedicine, Aarhus University, Aarhus, Denmark https://orcid.org/0000-0002-5055-7531
  • Jens Overgaard Department of Experimental Clinical Oncology, Aarhus University Hospital, Denmark https://orcid.org/0000-0002-0814-8179
  • Martin F Berthelsen Department of Clinical Medicine, Aarhus University, Denmark
  • Henrik Hager Department of Pathology, Aarhus University Hospital, Denmark https://orcid.org/0000-0002-3164-9190
  • Johan Bussink Department of Radiation Oncology, Radboud University, Nijmegen Medical Centre, Nijmegen, The Netherlands https://orcid.org/0000-0002-5751-4796
  • Kim V Hansen Department of Nuclear Medicine and PET Centre, Aarhus University Hospital, Denmark https://orcid.org/0000-0002-5665-3284
  • Steen Jakobsen Department of Nuclear Medicine and PET Centre, Aarhus University Hospital, Denmark
  • Jacob Kinggaard Lilja-Fischer Department of Experimental Clinical Oncology, Aarhus University Hospital, Denmark https://orcid.org/0000-0002-6400-6650
  • Ebbe Boedtkjer Department of Biomedicine, Aarhus University, Aarhus, Denmark
  • Mikkel H Vendelbo Department of Biomedicine, Aarhus University, Aarhus, Denmark; Department of Nuclear Medicine and PET Centre, Aarhus University Hospital, Denmark https://orcid.org/0000-0003-0431-2522

DOI:

https://doi.org/10.2340/1651-226X.2025.44002

Keywords:

hypoxia, radioresistance, image-guided radiotherapy, PET, FDG, FAZA

Abstract

Background and purpose: Quantification/mapping of tumor hypoxia may guide pretreatment decision-making in radiation oncology. Hypoxia-selective positron emission tomography (PET) tracers, like 18F-fluoroazomycin arabinoside (FAZA), allow assessment of hypoxia, but since hypoxia stimulates glycolysis, fluorodeoxyglucose (FDG) and hypoxia-PET may provide overlapping/similar information. Clinical dual-tracer PET studies are highly complex and remain inconclusive. Accordingly, we developed dual-tracer autoradiography techniques to allow high-resolution assessment of the spatial coupling of FAZA and 14C-2DG (FDG-analogue), without the time-separation and co-registration-related inaccuracies intrinsic to PET.

Patient/material and methods: Orthotopic lung adenocarcinomas were induced in CRISPR/Cas9 knock-in mice. Mammary adenocarcinomas developed spontaneously in transgenic mice overexpressing ErbB2 (Her2). Patient-derived-xenografts (PDX) were established in immunocompromised mice using biopsies from oropharyngeal cancer patients. Tumor growth was followed by MRI/Caliper measurements. Mice were administered with FAZA (~40 MBq)/14C-2DG (37 kBq)/pimonidazole and sacrificed. Tumor cryosections were analyzed for FAZA/14C-2DG using dual-tracer autoradiography followed by histological stainings. Complementary autoradiograms were co-registered and covered by a square-grid (0.5 × 0.5 mm), and Pearson correlation coefficients (R) were calculated.

Results/Interpretation: Hypoxic sub-volumes (FAZA/pimonidazole) were commonly present. A reasonable spatial overlap between FAZA and 14C-2DG was observed in most lung and oropharyngeal tumors with R typically exceeding 0.55. In the breast tumor model, the extent of overlap between FAZA and
14C-2DG varied widely with R ranging from 0.03 to 0.82, which may relate to intertumor mutational differences in this Her2+ oncogene-driven model. Our results suggest a putative role for FDG-PET to identify hypoxic foci and guide dose-escalation.

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Published

2025-08-30

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Busk, M., Thomsen, M. K., Overgaard, J., Berthelsen, M. F., Hager, H., Bussink, J., … Vendelbo, M. H. (2025). Dual-tracer autoradiographic analysis of glucose metabolism and hypoxia in orthotopic and PDX tumor models. Acta Oncologica, 64, 1152–1159. https://doi.org/10.2340/1651-226X.2025.44002

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Vol. 64 (2025): Acta Oncologica

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Copyright (c) 2025 Morten Busk, Martin K Thomsen, Jens Overgaard, Martin F Berthelsen, Henrik Hager, Johan Bussink, Kim V Hansen, Steen Jakobsen, Jacob Kinggaard Lilja-Fischer, Ebbe Boedtkjers, Mikkel H Vendelbo

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