SP8 - Pre-Clinical Validation

B. Lange (MPI-Molecular Genetics, Berlin)

We are applying a low passage mamma primary cell model to elucidate molecular changes of estrogen receptor (ER) signaling through systematically dissecting the pathway components by protein and mRNA profiling in combination with functional cellular assays. A second goal of this project is the evaluation of therapeutics on this low passage primary cell model that has the advantage of an in vitro system that is “as close as possible” to the patient.

• to generate of primary breast cancer cell lines
• to analyze the aberrations of estrogen receptor pathway in the primary cell model by mRNA and protein profiling in combination with functional cellular assays.
• to evaluate the relevance of anti-cancer therapies “as close as to the patient possible” through detailed analysis of the response to drug treatment in ER positive and ER negative cell lines (e.g., tamoxifen, parthenolide, rapomycin, gefitinib).
• to structurally characterize protein complexes relevant for oestrogen receptor signaling.

3D culture of established (MCF10A) und primary breast cancer cell lines (e.g., BCCL1) in basement membrane cultures, show formation of cyst like spheroids (acini) of MCF10A cells while the tested primary breast cancer cell lines show little or no tendency to organize spheroids under the same conditions. Confocal microscopy shows only few cells in the luminal space of the spheroid produced by the MCF10A cells (first image). In contrast, BCCL1 show (third image) single, spatially separated cells and nuclei (DNA staining with DAPI, blue). 1st image enlarged MCF10A spheroid, 2ed image MCF10A spheroids overview, 3rd image enlargement BCCL1 that form incoherent cellular 3D network 4thimage phase contrast microscopy overview over 3D BCCL1 culture that form incoherent cellular 3D network.