Pathology - Posters C
Titanilla Dankó1, Dorottya Moldvai1, Péter Sasvári2, Gábor Petővári1, Dániel Sztankovics1, Ildikó Krencz1, Regina Raffay1, Fanni Bugyi3, Lilla Turiák3, Anna Sebestyén1
1 Department of Pathology and Experimental Cancer Research, Semmelweis University, Budapest
2 Department of Physiology, Semmelweis University, Budapest
3 MS Proteomics Research Group, Research Centre for Natural Sciences, Budapest
Introduction. The development of preclinical models is necessary, as it is known that there may be significant differences between the outcomes of preclinical phase tests and the therapeutic effectivity. One of the aims of model developments is to create more complex test systems in order to model living tissue structures more realistically. This includes a new innovative technology, 3D bioprinting, which is a useful tool for creating tissue-like structures that can be cultured and tested in vitro.
Aims. Different models were applied for studying how two-dimensional (2D) and three-dimensional (3D) culturing conditions can affect metabolic features.
Methods. In our experiments, 5 model systems were established using ZR75.1 human breast cancer cell line: traditional 2D and various 3D (hanging drop/gel-embedded spheroids, and 3D bioprinted structures) in vitro cell cultures and in vivo xenografts. Mass spectrometric (MS) measurements and bioinformatic analysis using proteomic data were performed for metabolic comparative studies. Expression changes of proteins predicting metabolic differences in our experimental model systems were also tested using immunohistochemical (IHC) stainings.
Results. Based on the obtained results of proteomic data analysis, the samples have a normal distribution and are suitable for bioinformatic metabolic evaluation. Regarding our previous data on drug sensitivity differences in the 5 studied models, 2 proteins (GPx and ILK) were selected for further evaluation in protein expression studies. Differences in GPx and ILK expressions observed in the MS data analysis were validated in situ with IHC. Accordingly, 2D cell cultures can be characterized by reduced GPx protein expression level, in contrast, its expression was high in cells growing in 3D bioprinted structures and xenograft tumors.
Conclusion. All of these observations draw attention to the significant importance of the applied cell culturing methods, since these can affect the metabolic processes and influence the effects of pharmaceutical agents tested in different model systems. Consequently, the selection and the usage of models in preclinical trials are non-negligible factors from the point of view of drug sensitivity.
Funding. TKP2021-EGA-24, NKFI-FK-128404, NKFI-K-142799 (A.S.); ÚNKP-22-4-I-SE-12 (T.D.); ÚNKP-22-4-II-SE-9 (G.P.); EFOP-3.6.3-VEKOP-16-2017-00009 (D.S., D.M.)