Effect of gene electrotransfer on expression of cytosolic DNA sensors in tumor and normal cells in vitro and in vivo

Za vnos plazmidne DNA (pDNA) v celice ali tkiva lahko uporabimo različne dostavne metode. Ena izmed teh je elektroporacija, oz. genski elektroprenos (GEP) oz. elektrogenska terapija. Pri GEP celice izpostavimo zunanjemu električnemu polju z določeno jakostjo in trajanjem ter pri tem vplivamo na prepustnost membrane. Pri zdravljenju rakavih sprememb se pri elektrogenski terapiji uporabljajo predvsem geni, ki vplivajo na imunski odgovor ali angiogenezo tumorja. Pri predhodnjih poskusih smo na mišjem modelu z GEP uspeli regresijo tumorja doseči tudi pri kontrolni skupini, kjer smo uporabili le plazmidno DNA, brez terapevtskega gena. Upočasnjeno rast in regresijo tumorja po GEP plazmidne DNA brez terapevskega gena so opazili tudi v drugih raziskavah. Ker plazmidna DNA brez terapevtskega gena ne nosi zapisa za terapevtski protein, do regresije tumorja ni prišlo zaradi terapevtskega učinka. Predvidevamo, da plazmidna DNA z GEP vstopi v celico z endocitozo ali skozi pore v membrani, ki nastanejo po izpostavitvi celice električnim pulzom. Celice citosolno DNA zaznajo s pomočjo citosolnih senzorjev DNA v celici. Po vstopu pDNA v celico se ob njeni vezavi na senzorje DNA sproži kaskada reakcij znotrajceličnega signaliziranja, ki vodi do prepisa genov, ki kodirajo vnetne citokine in interferone. Poleg prepisovanja genov lahko pride tudi do sprožitve različnih oblik celične smrti in sicer piroptoze, nekroptoze in apotoze. Prisotnost senzorjev DNA so dokazali v imunskih in v nekaterih neimunskih celicah.For the entry of pDNA into the cells and tissues we can use different delivery methodes. One of them is electroporation, eg. gene electrotransfer (GET), eg. electrogene therapy in which the cells are exposed to electric pulses with specific intensities and durations in order to increase the permeability of the cell membrane. In cancer electrogene therapy, genes that affect the immune system or angioginesis of the tumor are used. In our laboratory, we noticed a regression of tumors after electrogene therapy of mice in control groups, where we used only pDNA without any therapeutic gene. In other laboratories tumor growth delay and regression of tumors after GET of pDNA was also determined. Because pDNA has no terapeutic genes, regression is not due to therapeutic effect. It was predicted that after GET, pDNA enters the cell with endocytosis or through pores, that are established in the membrane after exposure to the electric field. After the entery of pDNA into the cells, the DNA sensors can bind to it and this leads to dfferent signal pathways, resulting in transcription of genes coding proimmlamatory cytokines, interferons and can also lead to different triggered cell deaths, such as piroptosis and necroptosis. The presence of DNA sensors was shown in immune cells and some of the non-immune cells