Quantitative response assessment of combined immunotherapy in a murine melanoma model using multiparametric MRI

Abstract Background We assessed immunotherapy response in a murine melanoma model using multiparametric magnetic resonance imaging (mpMRI) features with ex vivo immunohistochemical validation. Methods Murine melanoma cells (B16-F10) were inoculated into the subcutaneous flank of n = 28 C57BL/6 mice...

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Main Authors: Maurice M. Heimer, Amra Cimic, Sandra Kloiber-Langhorst, Melissa J. Antons, Jennifer Stueckl, Heidrun Hirner-Eppeneder, Wolfgang G. Kunz, Olaf Dietrich, Jens Ricke, Felix L. Herr, Clemens C. Cyran
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Language:English
Published: SpringerOpen 2025-06-01
Series:European Radiology Experimental
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Online Access:https://doi.org/10.1186/s41747-025-00597-8
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author Maurice M. Heimer
Amra Cimic
Sandra Kloiber-Langhorst
Melissa J. Antons
Jennifer Stueckl
Heidrun Hirner-Eppeneder
Wolfgang G. Kunz
Olaf Dietrich
Jens Ricke
Felix L. Herr
Clemens C. Cyran
author_facet Maurice M. Heimer
Amra Cimic
Sandra Kloiber-Langhorst
Melissa J. Antons
Jennifer Stueckl
Heidrun Hirner-Eppeneder
Wolfgang G. Kunz
Olaf Dietrich
Jens Ricke
Felix L. Herr
Clemens C. Cyran
author_sort Maurice M. Heimer
collection DOAJ
description Abstract Background We assessed immunotherapy response in a murine melanoma model using multiparametric magnetic resonance imaging (mpMRI) features with ex vivo immunohistochemical validation. Methods Murine melanoma cells (B16-F10) were inoculated into the subcutaneous flank of n = 28 C57BL/6 mice (n = 14 therapy; n = 14 control). Baseline mpMRI was acquired on day 7 at 3 T. The immunotherapy group received three intraperitoneal injections of anti-PD-L1 and anti-CTLA-4 antibodies on days 7, 9, and 11 after inoculation. Controls received a volume equivalent placebo. Follow-up mpMRI was performed on day 12. We assessed tumor volume, diffusion-weighted imaging parameters, including the apparent diffusion coefficient (ADC), and dynamic-contrast-enhanced metrics, including plasma volume and plasma flow. Tumor-infiltrating lymphocytes (TIL; CD8+), cell proliferation (Ki-67), apoptosis (terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling, TUNEL), and microvascular density (CD31+) were assessed in a validation cohort of n = 24 animals for time-matched ex vivo validation. Results An increase in tumor volume was observed in both groups (p ≤ 0.004) without difference at follow-up (p = 0.630). A lower ADC value was observed in the immunotherapy group at follow-up (p = 0.001). Immunohistochemistry revealed higher TUNEL values (p < 0.001) and CD8+ TILs (p = 0.048) following immunotherapy, as well as lower tumor cell Ki-67 values (p < 0.001) and microvascular density/CD31+ (p < 0.001). Conclusion Lower tumor ADC, paired with higher intratumoral expression of CD8+ TIL, was observed five days after immunotherapy, suggestive of early immunological response. Ex vivo immunohistochemistry confirmed the antitumoral efficacy of immunotherapy. Relevance statement Compared to tumor size, diffusion-weighted MRI demonstrated potential for early response assessment to immunotherapy in a murine melanoma model, which could reflect changes in the tumor microenvironment and immune cell infiltration. Key Points No difference in tumor volume was observed between groups before and after therapy. Lower ADC values paired with increased CD8+ TILs were observed following immunotherapy. Ex vivo immunohistochemistry confirmed antitumoral efficacy of anti-PD-L1 and anti-CTLA-4 immunotherapy. Graphical Abstract
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spelling doaj-art-b523e15bf7924c22932b84a15714b2372025-08-20T03:20:59ZengSpringerOpenEuropean Radiology Experimental2509-92802025-06-019111310.1186/s41747-025-00597-8Quantitative response assessment of combined immunotherapy in a murine melanoma model using multiparametric MRIMaurice M. Heimer0Amra Cimic1Sandra Kloiber-Langhorst2Melissa J. Antons3Jennifer Stueckl4Heidrun Hirner-Eppeneder5Wolfgang G. Kunz6Olaf Dietrich7Jens Ricke8Felix L. Herr9Clemens C. Cyran10Department of Radiology, LMU University Hospital, LMU MunichDepartment of Radiology, LMU University Hospital, LMU MunichDepartment of Radiology, LMU University Hospital, LMU MunichDepartment of Radiology, LMU University Hospital, LMU MunichDepartment of Radiology, LMU University Hospital, LMU MunichDepartment of Radiology, LMU University Hospital, LMU MunichDepartment of Radiology, LMU University Hospital, LMU MunichDepartment of Radiology, LMU University Hospital, LMU MunichDepartment of Radiology, LMU University Hospital, LMU MunichDepartment of Radiology, LMU University Hospital, LMU MunichDepartment of Radiology, LMU University Hospital, LMU MunichAbstract Background We assessed immunotherapy response in a murine melanoma model using multiparametric magnetic resonance imaging (mpMRI) features with ex vivo immunohistochemical validation. Methods Murine melanoma cells (B16-F10) were inoculated into the subcutaneous flank of n = 28 C57BL/6 mice (n = 14 therapy; n = 14 control). Baseline mpMRI was acquired on day 7 at 3 T. The immunotherapy group received three intraperitoneal injections of anti-PD-L1 and anti-CTLA-4 antibodies on days 7, 9, and 11 after inoculation. Controls received a volume equivalent placebo. Follow-up mpMRI was performed on day 12. We assessed tumor volume, diffusion-weighted imaging parameters, including the apparent diffusion coefficient (ADC), and dynamic-contrast-enhanced metrics, including plasma volume and plasma flow. Tumor-infiltrating lymphocytes (TIL; CD8+), cell proliferation (Ki-67), apoptosis (terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling, TUNEL), and microvascular density (CD31+) were assessed in a validation cohort of n = 24 animals for time-matched ex vivo validation. Results An increase in tumor volume was observed in both groups (p ≤ 0.004) without difference at follow-up (p = 0.630). A lower ADC value was observed in the immunotherapy group at follow-up (p = 0.001). Immunohistochemistry revealed higher TUNEL values (p < 0.001) and CD8+ TILs (p = 0.048) following immunotherapy, as well as lower tumor cell Ki-67 values (p < 0.001) and microvascular density/CD31+ (p < 0.001). Conclusion Lower tumor ADC, paired with higher intratumoral expression of CD8+ TIL, was observed five days after immunotherapy, suggestive of early immunological response. Ex vivo immunohistochemistry confirmed the antitumoral efficacy of immunotherapy. Relevance statement Compared to tumor size, diffusion-weighted MRI demonstrated potential for early response assessment to immunotherapy in a murine melanoma model, which could reflect changes in the tumor microenvironment and immune cell infiltration. Key Points No difference in tumor volume was observed between groups before and after therapy. Lower ADC values paired with increased CD8+ TILs were observed following immunotherapy. Ex vivo immunohistochemistry confirmed antitumoral efficacy of anti-PD-L1 and anti-CTLA-4 immunotherapy. Graphical Abstracthttps://doi.org/10.1186/s41747-025-00597-8ImmunotherapyMelanomaMice (inbred C57BL)Multiparametric magnetic resonance imagingTumor microenvironment
spellingShingle Maurice M. Heimer
Amra Cimic
Sandra Kloiber-Langhorst
Melissa J. Antons
Jennifer Stueckl
Heidrun Hirner-Eppeneder
Wolfgang G. Kunz
Olaf Dietrich
Jens Ricke
Felix L. Herr
Clemens C. Cyran
Quantitative response assessment of combined immunotherapy in a murine melanoma model using multiparametric MRI
European Radiology Experimental
Immunotherapy
Melanoma
Mice (inbred C57BL)
Multiparametric magnetic resonance imaging
Tumor microenvironment
title Quantitative response assessment of combined immunotherapy in a murine melanoma model using multiparametric MRI
title_full Quantitative response assessment of combined immunotherapy in a murine melanoma model using multiparametric MRI
title_fullStr Quantitative response assessment of combined immunotherapy in a murine melanoma model using multiparametric MRI
title_full_unstemmed Quantitative response assessment of combined immunotherapy in a murine melanoma model using multiparametric MRI
title_short Quantitative response assessment of combined immunotherapy in a murine melanoma model using multiparametric MRI
title_sort quantitative response assessment of combined immunotherapy in a murine melanoma model using multiparametric mri
topic Immunotherapy
Melanoma
Mice (inbred C57BL)
Multiparametric magnetic resonance imaging
Tumor microenvironment
url https://doi.org/10.1186/s41747-025-00597-8
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