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Evaluating Tumor Hypoxia Radiosensitization Via Electron Paramagnetic Resonance Oxygen Imaging (EPROI)

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Abstract

Purpose

Tumor hypoxia contributes to aggressive phenotypes and diminished therapeutic responses to radiation therapy (RT) with hypoxic tissue being 3-fold less radiosensitive than normoxic tissue. A major challenge in implementing hypoxic radiosensitizers is the lack of a high-resolution imaging modality that directly quantifies tissue-oxygen. The electron paramagnetic resonance oxygen-imager (EPROI) was used to quantify tumor oxygenation in two murine tumor models: E0771 syngeneic transplant breast cancers and primary p53/MCA soft tissue sarcomas, with the latter autochthonous model better recapitulating the tumor microenvironment in human malignancies. We hypothesized that tumor hypoxia differs between these models. We also aimed to quantify the absolute change in tumor hypoxia induced by the mitochondrial inhibitor papaverine (PPV) and its effect on RT response.

Procedures

Tumor oxygenation was characterized in E0771 and primary p53/MCA sarcomas via EPROI, with the former model also being quantified indirectly via diffuse reflectance spectroscopy (DRS). After confirming PPV’s effect on hypoxic fraction (via EPROI), we compared the effect of 0 versus 2 mg/kg PPV prior to 20 Gy on tumor growth delay and survival.

Results

Hypoxic sarcomas were more radioresistant than normoxic sarcomas (p=0.0057, 2-way ANOVA), and high baseline hypoxic fraction was a significant (p=0.0063, Cox Regression Model) hazard in survivability regardless of treatment. Pre-treatment with PPV before RT did not radiosensitize tumors in the sarcoma or E0771 model. In the sarcoma model, EPROI successfully identified baseline hypoxic tumors. DRS quantification of total hemoglobin, saturated hemoglobin, changes in mitochondrial potential and glucose uptake showed no significant difference in E0771 tumors pre- and post-PPV.

Conclusion

EPROI provides 3D high-resolution pO2 quantification; EPR is better suited than DRS to characterize tumor hypoxia. PPV did not radiosensitize E0771 tumors nor p53/MCA sarcomas, which may be related to the complex pattern of vasculature in each tumor. Additionally, understanding model-dependent tumor hypoxia will provide a much-needed foundation for future therapeutic studies with hypoxic radiosensitizers.

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Data Availability

Research data are stored in an institutional repository and will be shared upon request to the corresponding authors.

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Acknowledgements

We thank Ketan Ghaghada, PhD (Texas Children’s Hospital) for providing liposomal iodine and protocols for cone-beam computed tomography acquisition. We also thank O2M team members, Eliyas Siddiqui, and University of Chicago’s senior technician Eugene Barth for their support with EPROI experiment training and data processing.

Funding

This work is funded by the O2M Technologies and National Institutes of Health (NCI R44CA224840, PI: Kotecha; NCI R41OD026688, PI: Ramanujam). YMM received support from a K08 Award from National Institute of Dental and Craniofacial Research (K08-DE029887). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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Authors and Affiliations

  1. Department of Radiation Oncology, Duke University Medical Center, Durham, NC, USA

    Ashlyn G. Rickard, Yvonne M. Mowery, Alex Bassil, Nerissa T. Williams, Theresa Charity, Rafaela Belloni, Rico Castillo & Gregory M. Palmer

  2. Department of Head and Neck Surgery & Communication Sciences, Duke University Medical Center, Durham, NC, USA

    Yvonne M. Mowery

  3. Division of Laboratory Animal Resources, Duke University School of Medicine, Durham, NC, USA

    Douglas C. Rouse

  4. Department of Biomedical Engineering, Duke University, Durham, NC, USA

    Brian Crouch & Nimmi Ramanujam

  5. Zenalux Biomedical, Inc, Durham, NC, USA

    Daniel Stevenson

  6. Department of Radiology, Duke University Medical Center, Durham, NC, USA

    Stephanie Blocker

  7. Department of Radiation and Cellular Oncology, University of Chicago, Chicago, IL, USA

    Boris Epel

  8. O2M Technologies LLC, Chicago, IL, USA

    Boris Epel & Mrignayani Kotecha

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  1. Ashlyn G. Rickard
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  2. Yvonne M. Mowery
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Contributions

YMM, BE, MK, and GMP contributed to the conception and design of the experiment. AGR, YMM, AB, NTW, TC, RB, SB, and RC acquired and analyzed animal data. AGR and GMP were responsible for the statistical analyses. BE provided oversight in EPROI acquisition and analysis. DCR trained and oversaw jugular vein cannulation. BC, NR, DS, and GMP oversaw and analyzed DRS data. All authors contributed to the final manuscript.

Corresponding authors

Correspondence to Yvonne M. Mowery, Mrignayani Kotecha or Gregory M. Palmer.

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Conflict of Interest

GMP, BC, NR, DS, and Duke University have financial interest in Zenalux Biomedical, Inc. which is commercializing the Zenascope. BE and MK have financial interest in O2M Technologies, LLC. All other authors declare no competing interests.

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Rickard, .G., Mowery, Y.M., Bassil, A. et al. Evaluating Tumor Hypoxia Radiosensitization Via Electron Paramagnetic Resonance Oxygen Imaging (EPROI). Mol Imaging Biol (2023). https://doi.org/10.1007/s11307-023-01855-0

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