Immunohistochemical Evaluation of Candidate Biomarkers for Fluorescence-Guided Surgery of Myxofibrosarcoma Using an Objective Scoring Method

Zeger Rijs; Esther Belt; Gijsbert M Kalisvaart; Cornelis F M Sier; Peter J K Kuppen; Arjen H G Cleven; Alexander L Vahrmeijer; Michiel A J van de Sande; Pieter B A A van Driel

doi:10.3390/biomedicines11030982

Immunohistochemical Evaluation of Candidate Biomarkers for Fluorescence-Guided Surgery of Myxofibrosarcoma Using an Objective Scoring Method

Biomedicines. 2023 Mar 22;11(3):982. doi: 10.3390/biomedicines11030982.

Authors

Affiliations

¹ Department of Orthopedic Surgery, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands.
² Department of Radiology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands.
³ Department of Surgery, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands.
⁴ Percuros BV, Zernikedreef 8, 2333 CL Leiden, The Netherlands.
⁵ Department of Pathology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands.
⁶ Department of Pathology, University Medical Center Groningen, 9700 RB Groningen, The Netherlands.
⁷ Department of Orthopedic Surgery, Isala Hospital, 8025 AB Zwolle, The Netherlands.

Abstract

Introduction: Myxofibrosarcoma (MFS) is the most common soft-tissue sarcoma subtype in elderly patients. Local recurrence (LR) remains a major concern as the lack of intraoperative guidance and an infiltrative growth pattern with long, slender tails hamper surgeons' ability to achieve adequate resection margins for MFS. Fluorescence-guided surgery (FGS) could overcome this concern by delineating tumor tissue during surgery. One of the most important steps to successful FGS is to define a tumor-specific biomarker that is highly overexpressed in tumor tissue while low or absent in adjacent healthy tissue. The aim of this study is to evaluate the expression of eight previously selected promising biomarkers for FGS in MFS tissue samples with adjacent healthy tissue using immunohistochemistry (IHC).

Methods: The following eight biomarkers were stained in seventeen paraffin-embedded MFS samples: tumor endothelial marker-1 (TEM-1, also known as endosialin/CD248), vascular endothelial growth factor receptor-1 (VEGFR-1, also known as Flt-1), vascular endothelial growth factor receptor-2 (VEGFR-2, also known as Flk1), vascular endothelial growth factor-A (VEGF-A), epidermal growth factor receptor (EGFR), insulin-like growth factor-1 receptor (IGF-1R), platelet derived growth factor receptor-α (PDGFR-α), and cluster of differentiation 40 (CD40, also known as TNFRSF5). A pathologist specializing in sarcoma annotated the margin between the tumor and adjacent healthy tissue in each MFS tissue sample. Subsequently, we used an objective IHC scoring method to assess and compare the difference in staining intensity between the tumor and adjacent healthy tissue, which is crucial for the use of FGS.

Results: TEM-1, VEGF-A, and PDGFR-α stained all MFS tumors, while the other biomarkers did not show expression in all MFS tumors. Ultimately, TEM-1 was identified as the most suitable biomarker for FGS in MFS based on higher tumor-to-background (TBR) staining intensity compared to VEGF-A and PDGFR-α, regardless of preoperative therapy.

Conclusion: TEM-1-targeted FGS tracers should be further investigated to optimize MFS treatment.

Keywords: fluorescence-guided surgery; immunohistochemistry; molecular imaging; soft tissue sarcoma; tumor endothelial marker-1.

Grants and funding

C.F.M.S. was supported by the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreements No. 734684 (CHARMED), 872391 (CONCRETE), and PRISAR2 (872860). Part of the PhD program of Z.R. is funded by donations from Stichting Team Westland (STW).