Manipulacja oporności lekowej w komórkach nowotworowych poprzez nanosekundowe, asymetryczne sekwencje impulsów

Dofinansowano ze Środków Budżetu Państwa

  • Granty Narodowego Centrum Nauki

Nazwa programu: 


Całkowita wartość zadania: 

1 004 370,00 zł

Wartość finansowania: 

1 004 370,00 zł

Opis projektu: 

Electroporation (EP) is a promising method of wide applications. This technology enables effective drug and gene delivery which can be implemented in therapeutic protocols, food preservation, cosmetology, or genetic engineering. Up to now, there are few types of electroporation determined by the range of used electric pulses. In this project, asymmetric nanosecond pulsed electric field (AnsPEF) will be used and carefully examined. AnsPEF is a completely novel method used here for the first time. The project will involve the development of a new platform for high power sub-microsecond asymmetric bipolar pulse forming based on the latest silicon-carbide solid-state electronic technologies. The newly developed generator supporting loads up to 100 A and capable of inducing up to 50 kV/cm electric fields in the sample will enable the study of the new electroporation phenomena in the whole sub-microsecond (100 – 900 ns) asymmetric pulse range, which was not performed before. Lastly, to ensure multiparametric coverage the new platform will support the high frequency (MHz range) operation mode and the resulting pulse distortion problems due to transient processes will be effectively solved indicating the transdisciplinary novelty of the objectives. There will be verified what is the mechanism of action of this new technology in the cancer cell model including also cells with drug resistance. There is expected that the high energy asymmetric ns pulses elicit proportionally stronger effects on intracellular membranes than longer and standard pulses and in consequence may be more effective in drug and gene delivery. The drug resistance in vitro model was intentionally included in the research, cause the last data proved that this type of cell is more sensitive to electrical pulses. Moreover, gaining insight of the mechanisms altering drug resistance is important to develop more effective future therapeutic approaches. The application of ultra-short asymmetric electrical pulses may affect drug resistance phenomena, activate mechanisms responsible for oxidative stress, and intracellular pathways inducing increased permeabilization of cell membranes and finally cell death. Although the electroporation with longer unipolar pulses, used in standard protocols, was well described, the mechanisms of asymmetric nsPEF were not investigated and require thorough evaluation. The purpose of the project is an understanding of the mechanisms of action of AnsPEF method in the cancer cell model. Moreover, protocols AnsPEF based will be optimized for the effective drug and gene delivery for effective future applications. In the project the following research hypotheses will be addressed: The application of asymmetrical waveform of electric nanosecond pulses: • induce effective cancer cell membrane permeabilization for drug and gene delivery • inhibit the activity of drug resistance pumps • alternate viability of cancer cells and induce free radical processes • will be efficient in electrotransfer of large molecules.