PhD Scientific Days 2022

Budapest, 6-7 July 2022

Pathology and Oncology I. (Poster discussion will take place in the Aula during the Coffee Break)

Drug Sensitivity Differences in In Vitro Model Systems

Titanilla Dankó1, Gábor Petővári1, Regina Raffay1, Dorottya Moldvai1, Dániel Sztankovics1, Ildikó Krencz1, Krisztina Sipos1, Enikő Vetlényi1, Anna Sebestyén1
1 Semmelweis University, Department of Pathology and Experimental Cancer Research, Budapest

Text of the abstract

Introduction
Traditional 2D cell culturing could not represent the heterogeneity and complexity of the in vivo growing tumors, especially the metabolic heterogeneity of cancer tissues. This could be one reason for the failures of potential therapeutic drugs which are usually started to be selected and developed in in vitro models. There are several 3D culturing possibilities and new 3D bioprinted in vitro tissue mimetic cultures which are being developed for tissue, cell culturing experimental tests.
Aims
For comparing the effects of culturing conditions on drug sensitivity screening effectivity and metabolic profiles/characteristics, we examined 2D and 3D cell maintaining methods using breast cancer cell lines in vitro and in vivo.
Methods
We applied traditional hanging drop technique, ultra-low-attachment plates and established a 3D bioprinted model for culturing human breast cancer cell lines. In vitro cellular proliferation was monitored using cell counting, Alamar blue and sulforhodamine B tests after rapamycin, doxycycline and doxorubicin mono- and co-treatments. The in vitro sensitivity differences of 2D and 3D cultures were compared to the in vivo detected tumor growth. Wes Simple technique and immunohistochemical stainings were used to analyze the mTOR and metabolic enzyme activities of 2D, 3D and in vivo models.
Results
We verified that our developed 3D bioprinting method is suitable for creating a more complex, tissue-like environment for maintaining living cells as well as this could be applied for in vitro drug sensitivity testing. Using different models of breast cell lines, we showed that changes in metabolic activity and spatial heterogeneity of tumor tissue are important in therapy resistance. Additionally, we highlighted that rapamycin+doxycycline/doxorubicin multi-targeting may be effective in the treatment of malignancies.
Conclusion
The applied model systems of drug pre-selection processes could modify the cellular response. Therefore, there is a need for developing accurate model systems to mimic the living conditions more preciously in experimental systems/early phase of drug screening. 3D bioprinting is a new innovation that could also support the comprehension of the background adaptation processes and help find potential better drug candidates.
Funding
NKFI-FK128404, EFOP-3.6.3-VEKOP-16-2017-00009, Stephen W. Kuffler Foundation