PO_I_P: Pathology and Oncology I. Posters
Titanilla Dankó1, Gábor Petővári1, Dániel Sztankovics1, Dorottya Moldvai1, Regina Raffay1, Patrícia Kóczán1, Péter Lőrincz2, Tamás Visnovitz3, Viktória Zsiros4, Gábor Barna1, Ágnes Márk1, Ildikó Krencz1, Anna Sebestyén1
1 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, H-1085 Budapest, Üllői út 26. Hungary
2 Department of Anatomy, Cell and Developmental Biology, Eotvos Lorand University, H-1117, Budapest Pázmány Péter sétány 1/c. Hungary
3 Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, H-1089, Nagyvárad tér 4. Hungary
4 Department of Anatomy, Histology and Embryology, Semmelweis University, Budapest, H-1094, Tűzoltó utca 58. Hungary
Metabolic remodeling is a characteristic of cancer cells as a response to treatments. Besides promoting cell growth and survival, the activation of mTOR pathway can also lead to developing resistance mechanism or metastasis formation. In addition, autophagy has a double-edged role as it can either maintain cellular homeostasis or stimulate tumorigenesis in stress.
As doxycycline (antibiotic agent) has an off-target effect on mitochondrial processes, we aimed to study whether its combination with rapamycin (mTOR inhibitor) is potentially more effective in treatments.
Rapamycin + doxycycline was tested in several cell lines in vitro, and the mechanism of action was examined using human breast cancer models. Alamar blue and sulforhodamine B tests were applied and the impact of long-term mono- and combination treatments on cell and tumor growth was also monitored in in vitro and in vivo. The induced cell death was detected by flow cytometry and caspase-3 activity measurements. mTOR-, autophagy-, and necroptosis-related proteins were analyzed by WES, Western blot and IHC. Treatment-induced morphological alterations were examined by fluoresce and transmission electron microscopy.
Rapamycin + doxycycline combination could decrease the cell proliferation capacity in the majority of the investigated cell lines in vitro. Continuous rapamycin + doxycycline addition could reduce cell and tumor growth significantly. The studied combination did not cause apoptosis, necrosis or necroptosis, but altered the expressions of LC3 and p62 which were associated with autophagy induction. Lowered intensity of MitoTracker and TOM20 staining suggested that mitophagy could be induced after rapamycin + doxycycline treatment. In parallel, rapamycin + doxycycline combination increased the sign of autophagy (mitophagy)-related alterations at tissue level.
A novel therapeutic strategy could be to co-target mitochondria and mTOR pathway to influence metabolic rewiring and inhibit metabolic adaptation in tumor cells.
NKFI-FK-128404, National Bionics program-ED_17-1-2017-0009, STIA-KFI2020, VEKOP-2.3.2-16-2016-00002, OTKA120237, PPD-222/2018
Semmelweis University, Doctoral School of Pathological Sciences