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. 2023 Jul 13;15(14):3602.
doi: 10.3390/cancers15143602.

Advancing Cancer Therapy Predictions with Patient-Derived Organoid Models of Metastatic Breast Cancer

Cansu E Önder  1 , Teresa J Ziegler  1 , Ronja Becker  1 , Sara Y Brucker  1   2 , Andreas D Hartkopf  2 , Tobias Engler  2 , André Koch  1   2
Affiliations

Advancing Cancer Therapy Predictions with Patient-Derived Organoid Models of Metastatic Breast Cancer

Cansu E Önder et al. Cancers (Basel). .

Abstract

The poor outcome of metastasized breast cancer (BC) stresses the need for reliable personalized oncology and the significance of models recapitulating the heterogeneous nature of BC. Here, we cultured metastatic tumor cells derived from advanced BC patients with malignant ascites (MA) or malignant pleural effusion (MPE) using organoid technology. We identified the characteristics of tumor organoids by applying immunohistochemistry and mutation analysis. Tumor organoids preserved their expression patterns and hotspot mutations when compared to their original metastatic counterpart and are consequently a well-suited in vitro model for metastasized BC. We treated the tumor organoids to implement a reliable application for drug screenings of metastasized cells. Drug assays revealed that responses are not always in accord with expression patterns, pathway activation, and hotspot mutations. The discrepancy between characterization and functional testing underlines the relevance of linking IHC stainings and mutational analysis of metastasized BC with in vitro drug assays. Our metastatic BC organoids recapitulate the characteristics of their original sample derived from MA and MPE and serve as an invaluable tool that can be utilized in a preclinical setting for guiding therapy decisions.

Keywords: ascites; breast cancer; cancer biology; drug response; metastasis; organoid culture; patient-derived organoids; personalized medicine; pleural effusion.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Figures

Figure 1
Figure 1
Establishing a biobank of breast cancer (BC) organoids derived from pleural effusion and ascites. (A) Brightfield images of metastatic BC patient-derived organoid (MBC-PDO) #07 in passage (P)0, P1, P5, and P10. Morphology was preserved throughout the whole culturing period. Scale bar: 500 µm. (B) Brightfield images of organoid lines displaying different phenotypes. The first image with grape-like morphology illustrates the ILC sample MBC-PDO #02, and images with dense, smooth, and rough structures illustrate NST samples of MBC-PDO #03–#07. Scale bar: 100 µm.
Figure 2
Figure 2
Histological characterization of pleural BC cells and organoids derived from pleural effusion. Histology (H&E staining) and receptor status of metastasized liver tissue, pleural cells, and organoids (P3) of MBC-PDO #03 are shown. Organoids consist exclusively of epithelial cells, while tissues often show tumor epithelium ringed by mesenchymal cells. The receptor status is maintained in the organoids. Scale bar: 100 µm.
Figure 3
Figure 3
In vitro drug response assays of organoid lines. (A) Overview of drugs used in the assays and their targets. Inhibitors Afatinib, Lapatinib, Neratinib, and Tucatinib target the HER2 receptor intracellularly. Alpelisib and Pictilisib target PI3K, while Ipatasertib and Capivasertib aim for AKT. Everolimus inhibits mTORC1, whereas Abemaciclib and Palbociclib target CDK4/6 in the nucleus. Olaparib, which is administered to BC patients with a BRCA1/2 mutation/deletion, inhibits PARP. Gemcitabine enters the cell through nucleoside transporters and is phosphorylated in three steps into Gemcitabine triphosphate, which is incorporated into the DNA and leads to the termination of DNA synthesis. Paclitaxel binds and stabilizes β-tubulin, upon which depolymerization is blocked. This affects the mitotic spindle assembly, chromosome segregation, and mitosis. (B) Heat map of relative cell viability of organoids treated with various drugs at specific concentrations for 4 days (10 µM Afatinib, 1 µM Neratinib, 4 µM Lapatinib, 10 µM Tucatinib, 10 µM Alpelisib, 1 µM Pictilisib, 1 µM Ipatasertib, 1 µM Capivasertib, 100 nM Everolimus, 4 µM Abemaciclib, 10 µM Palbociclib, 10 µM Olaparib, 100 nM Gemcitabine, 10 nM Paclitaxel). Values were normalized to control treatment with 0.1% DMSO (set to 100%). Red fields indicate sensitive responses, while blue fields present less-responding lines. Experiments were performed in technical triplicates. (C) Drug response curves depict organoid viabilities after 4 days of treatment with Lapatinib, Alpelisib, Capivasertib, Abemaciclib, and Paclitaxel. Error bars representing standard deviation (SD) of three independent experiments were removed for better presentation. IC50 values of drug response curves are shown in the adjacent table.
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