20.500.12556/RUL-110025 Imunoterapija raka: regulacija T-celic z dizajniranim transkripcijskim faktorjem NFAT Cancer immunotherapy: T-cell regulation via engineered transcription factor NFAT Razvoj terapevtskih postopkov rakavih obolenj se zaradi kompleksnosti slednjih nagiba k usmerjenim in personaliziranim adaptivnim celičnim terapijam. Mednje sodijo celice CD19 CAR-T, pri čemer pacientovim limfocitom T vstavimo receptor CAR, specifičen za B-limfocitni antigen CD19. Terapija je uspešna pri B-celičnih malignih obolenjih, vendar so prisotni stranski učinki (SIRS, CRS) zaradi katerih je potrebno uravnavanje aktivnosti celic CAR-T. Z namenom regulacije celic CAR-T smo na osnovi transkripcijskega faktorja NFAT in heterodimerizacijskih domen ABI-PYL1, DmrA-DmrC in GAI-GID1 pripravili regulatorni sistem, ki celice aktivira ali represira. V celicah HEK293T in Jurkat smo potrdili, da ob dodatku ustreznih induktorjev (abscizinska kislina, rapamicin in giberelin) pride do heterodimerizacije konstruktov in da se NFAT učinkovito veže na vezavna mesta za DNA, pri čemer zanje tekmuje z endogenim NFAT. Hkrati smo pokazali, da heterodimerizacijski sistemi robustno aktivirajo celice Jurkat z receptorjem CD19-CAR ob prisotnosti celic, ki izražajo tarčni antigen (celice Raji). In recent years, cancer immunotherapy has been leaning towards personalized, T-cell mediated therapy, due to the complex background of the disease. CD19 CAR-T cells represent a cure for hematologic malignancies. The therapy, albeit successful, does not avoid side effects such as SIRS and CRS and thus requires a development of a robust and reliable regulation of CAR-T cells. With CAR-T regulation in mind, we developed a regulatory system, based on transcription factor NFAT and heterodimerization domains ABI-PYL1, DmrA-DmrC, GAI-GID1, which either activates or represses the cells in question. We show that in the presence of chemical inductors (abscisic acid, rapamycin and gibberellin) the constructs undergo heterodimerization and that engineered NFAT, competing with endogenic NFAT, binds to its regulatory site of both reporter gene (shown in HEK293-T cells) or IL-2 (shown in Jurkat cells). We also confirm, that heterodimerization systems work in cells with expressed CD19-CAR and in presence of target antigen on the surface of Raji cells. CAR-T imunoterapija raka heterodimerizacija rapamicin giberelin abscizinska kislina CAR-T cancer immunotherapy heterodimerization rapamycin gibberellin abscisic acid true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2019-09-11 08:55:06 2019-09-11 08:55:07 2022-08-22 03:41:10 0000-00-00 00:00:00 2019 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 6674 1538428355 Malensek_Spela_-_Imunoterapija_raka-_regulacija_T-celic_z_dizajniranim_transkripcijskim_faktorje.pdf Malensek_Spela_-_Imunoterapija_raka-_regulacija_T-celic_z_dizajniranim_transkripcijskim_faktorje.pdf 1 7BEF6AAD9EBFBD8DBFB4D9A6335C3B48 5ebbb80deed584a727527611da6a0b7830c32a8ba8496de95f3fb1c814786cc7 03e30f5c-a1b7-11eb-a523-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=121642 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0