Intraoperative radiotherapy for breast cancer treatment efficiently targets the tumor bed preventing breast adipose stromal cell outgrowth

Stefanie Uhlig, Anne Wuhrer, Sebastian Berlit, Benjamin Tuschy, Marc Sütterlin, Karen Bieback, Stefanie Uhlig, Anne Wuhrer, Sebastian Berlit, Benjamin Tuschy, Marc Sütterlin, Karen Bieback

Abstract

Objectives: Mesenchymal stromal cells (MSC) in bone marrow have been shown to be radioresistant, which is related to pronounced DNA repair mechanisms. Intraoperative radiotherapy (IORT) during breast-conserving surgery for early breast cancer is an innovative technique applying low energy x‑ray to the tumor bed immediately after removal of the tumor. IORT is considered to reduce the risk of local tumor recurrence by directly targeting cells of the tumor bed and altering the local microenvironment. Aim of this study was to investigate whether IORT affects the outgrowth potential of breast adipose tissue-derived MSC (bASC) as part of the tumor bed.

Materials and methods: After surgical tumor resection, biopsies of the tumor bed were taken before (pre IORT) and after IORT (post IORT) and processed applying well-established protocols for ASC isolation and characterization.

Results: In all, 95% of pre IORT tumor bed samples yielded persistently outgrowing bASC with typical ASC characteristics: fibroblastoid morphology, proliferation, adipogenic and osteogenic differentiation and ASC surface marker expression. However, none of the post IORT samples yielded persistent outgrowth of bASC.

Conclusions: After breast-conserving surgery, approximately 90% of local recurrences emerge in close proximity to the initial tumor bed, potentially reflecting a significant contribution of the tumor bed to relapse. Our data show that IORT, besides the proven effect on breast cancer cells, efficiently modifies the tumor environment by having an impact on tumor bed bASC. This effect on tumor bed stromal cells might contribute to reduce the risk of tumor relapse and metastases.

Keywords: Adipose stromal cells; Breast cancer; Intraoperative radiotherapy; Mesenchymal stromal cells; Tumor bed.

Conflict of interest statement

S. Uhlig, A. Wuhrer, S. Berlit, B. Tuschy, M. Sütterlin and K. Bieback declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Characterization of breast-derived adipose stromal cells (bASC). a Phase contrast photomicrographs in p0 of a pre and post IORT sample. The typical mesenchymal stromal cells(MSC)-like morphology is apparent in the pre intraoperative radiotherapy (IORT) sample. All cells attaching from the post IORT sample show a senescent phenotype: cells lose their fibroblastoid shape, become flat and huge with bundles of stress fibers. To allow for better comparison, contrast and brightness were slightly adjusted. b Exemplary figures of adipogeneic (ADA, Oil Red O stain) and osteogenic (ODA, von Kossa stain) differentiation results displaying the negative controls (top rows) and the adipogenic/osteogenic-differentiated samples (bottom rows). c, d Flow cytometric assessment of MSC markers and e, f markers indicating contamination and other markers used to characterize putative subpopulations of MSC. Data are expressed as percent positive cells or mean fluorescence values MFI (median FL-A). No differences were seen; thus data are merged from all conditions. MFI mean fluorescence intensity, ADA adipogenic differntiation assay, ODA osteogneic differentiation assay

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Source: PubMed

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