Patient-derived scaffolds as a drug-testing platform for endocrine therapies in breast cancer

Three-dimensional cell culture platforms based on decellularised patient-based microenvironments provide in vivo-like growth conditions allowing cancer cells to interact with intact structures and components of the surrounding tissue. A patient-derived scaffold (PDS) model was therefore evaluated as a testing platform for the endocrine therapies (Z)-4-Hydroxytamoxifen (4OHT) and fulvestrant as well as the CDK4/6-inhibitor palbociclib, monitoring the treatment responses in breast cancer cell lines MCF7 and T47D adapted to the patient-based microenvironments. MCF7 cells growing in PDSs showed increased resistance to 4OHT and fulvestrant treatment (100- and 20-fold) compared to 2D cultures. Quantitative PCR analyses of endocrine treated cancer cells in PDSs revealed upregulation of pluripotency markers further supported by increased self-renewal capacity in sphere formation assays.

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FIG: Size optimization of patient-derived scaffolds to enable maximum number of downstream experiments. (a) A schematic overview depicting the different steps from tumor collection, experimental procedures to data analysis. (b) Viability of MCF7 cells after three weeks of growth in patient-derived scaffolds (PDSs) with three different thickness (50, 100 and 150 µm) using alamar blue. Data is represented relative to alamar blue levels of 50 µm slices. Mean + SEM is shown, n = 3 PDSs. One-Way ANOVA with Tukey’s multiple comparison test was done between all the groups combinations (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001). (c) Gene expression of MCF7 cells cultured in PDSs with different thicknesses relative to the expression in two-dimensional (2D) cultures and expressed in log2-scale. Mean + SEM is shown, n = 3 PDSs. (d) Image showing a representative hematoxylin and eosin staining of a 4 µm cross-section from a 150 µm PDS slice cultured with MCF7 cells after three weeks of growth. Bar represents 100 µm. (e) Bar graph demonstrating gene expression analysis of MCF7 cells after growth in 6 different PDSs compared to 2D cultures and expressed in log2-scale. Individual PDSs are indicated by colors and each dot represent a PDS slice. Mean + SEM is shown, n = 5–6. Two-Way ANOVA with Tukey’s multiple comparison test was done between all the groups combinations (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001). Additional genes are included in Supplementary Fig.

When comparing different 3D growth platforms including PDS, matrigel, gelatin sponges and 3D-printed hydrogels, 3D based cultures showed slightly varying responses to fulvestrant and palbociclib whereas PDS and matrigel cultures showed more similar gene expression profiles for 4OHT treatment compared to the other platforms. The results support that the PDS technique maximized to provide a multitude of smaller functional PDS replicates from each primary breast cancer, is an up-scalable patient-derived drug-testing platform available for gene expression profiling and downstream functional assays.

Gustafsson, A., Garre, E., Leiva, M.C. et al. Patient-derived scaffolds as a drug-testing platform for endocrine therapies in breast cancer. Sci Rep 11, 13334 (2021). https://doi.org/10.1038/s41598-021-92724-9

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