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A7103 - Characterization and Pharmacological Intervention of Tumor-Microenvironmental Factors and Cancer Cell Growth in Co-Cultures of Fresh Human Lung Tissue and Patient-Derived, Disseminated Cancer Cells
Author Block: S. Konzok1, S. Dehmel1, C. Werno2, P. Braubach3, G. Warnecke3, P. Zardo3, D. Jonigk3, B. Polzer2, K. Weidele2, C. Klein4, A. Braun1, K. Sewald1; 1Fraunhofer ITEM, Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Centre for Lung Research (DZL), Hannover, Germany, 2Fraunhofer ITEM, Regensburg, Germany, 3Hannover Medical School (MHH), Biomedical Research in Endstage and Obstructive Lung Disease Hannover (BREATH), Member of the German Centre for Lung Research (DZL), Hannover, Germany, 4University Hospital Regensburg, Regensburg, Germany.
The main cause of death in cancer is not the primary tumor itself but the transgression towards new organs as part of the invasive metastasis cascade. Metastases are characterized by high cellular heterogeneity and genetic disparity, leading to complex processes, which can only partly be reflected in most in-vitro models due to a lacking human microenvironment.
Anti-tumor drugs cisplatin, bevacizumab and vemurafenib were used to modulate different aspects of tumorigenesis. Effects on tumor growth reduction and modulation within its natural microenvironment in solid tumors were determined by preparing fresh human tumor tissue slices. Co-cultures of fresh human lung tissue slices with either allogenic GFP-labeled MDA-MB-231 or patient-derived, disseminated and GFP-labeled melanoma cells were treated analogously to the tumor slices to gain insight into first steps in the metastatic process.
Cancer cells integrate into the healthy lung tissue and proliferate within the first 24 hours of co-culture [6fold increase]. Neoangiogenetic biomarker VEGF was elevated 3.5fold in lung tissue co-cultures with MDA-MB-231 cells and 5.4fold in lung tumor slices after 48h. Treatment with bevacizumab [200 μg/mL] suppressed VEGF-release up to 24fold in co-cultures and up to 25fold in lung tumor tissue slices after 48h. Consequently, supernatants of these treatments also showed impaired endothelial cells migration by up to 81% in co-cultures and up to 83% in tumor slices. Other tumor biomarkers like GM-CSF were also highly elevated [13fold after 24h] in co-cultures compared to healthy tissue controls. Cisplatin treatment [50μM] led to a decline of viability and reduced cancer cell number in co-cultures by up to 37.5% and in tumor slices up to 48.7% after 72h. Cancer cell-invaded lung tissue and tumor slices were less sensitive in regards to their respective anti-cancer drug efficacy than the 2D culture. To address patient-specific genetic disparities, co-cultures of lung tissue and disseminated melanoma cells were performed with both patient-derived cells carrying driver BRAF mutation V600E and non-mutated melanoma cells. Treatment with vemurafenib [50µM], which interrupts the B-Raf/MEK/ERK pathway in V600E mutants, led to a 71% decrease of V600E cancer cells after 48 hours whereas non-mutated cells showed no significant cancer cell decrease.
Here we modulate cancer cell proliferation, growth and mediator concentrations in human lung tissue, showing that tumor cells in solid tumors and freshly seeded into healthy lung tissue are sensitive to tumor treatments ex vivo. This work aims to translate cancer drug efficacy data from animals towards humans to optimize clinical trial outcome.