Učinki antagonistov tirozin kinaznega receptorja TGF-ßRI in kemokinskega receptorja CCR5 v 3D celičnih modelih glioblastoma

Pinter, Eva (avtor), Breznik Vittori, Barbara (mentor), Novak, Metka (komentor)

Pri možganskem tumorju glioblastomu (GBM) se srečujemo s številnimi izzivi, med drugim z odpornostjo na standardno zdravljenje z obsevanjem in kemoterapijo ter imunosupresivnim tumorskim mikrookoljem (TME). Rakave celice izločajo imunosupresivne molekule kot sta transformirajoči rastni dejavnik beta (TGF-β) in C-C kemokinski ligand 5 (CCL5), ki privabljata in reprogramirata imunske celice, da delujejo v prid rasti tumorja. Razvoj novih terapevtskih strategij gre v smeri kombinacije različnih pristopov, ki ciljajo TME. Namen magistrske naloge je bil ovrednotiti učinke antagonistov serin/treonin kinaznega receptorja TGF-βRI in kemokinskega receptorja CCR5 na viabilnost ter invazijo tridimenzionalnih (3D) celičnih modelov GBM. Poleg tega smo želeli ugotoviti, ali ciljanje teh citokinskih poti poveča občutljivost sferoidov in organoidov glioblastoma (GBO) na standardno terapijo (ST). Primerjava viabilnosti in invazije sferoidov bolnikov z GBM je pokazala izrazito heterogenost odziva na inhibitor TGF-βRI kot monoterapijo in v kombinaciji s ST, tako med bolniki kot tudi znotraj istega bolnika med sferoidi, pripravljenimi iz diferenciranih celic glioblastoma (dGBM) in glioblastomskih matičnih celic (GSC). Pri dveh od šestih sferoidov se je po inhibiciji TGF-βRI z galunisertibom (GAL) povečala občutljivost na ST. Analiza prenosa western je v sferoidih GBM pokazala okrnjeno od SMAD (angl. suppressor of mother against decapentaplegic)-odvisno signalno pot TGF-β, kar nakazuje na obstoj kompenzacijske poti, ki uravnava proces invazije. Izražanje TGF-βRI se je po inhibiciji z GAL pri nekaterih sferoidih povečalo, pri drugih zmanjšalo, medtem ko je inhibicija signalne poti TGF-β v vseh sferoidih dGBM/GSC povzročila aktivacijo od SMAD-neodvisne signalne poti ERK. Maravirok (MRV), antagonist CCR5, kot monoterapija ni imel učinka na viabilnost GBO, je pa povečal invazijo GBO, kar bi lahko bila posledica aktivacije kompenzacijske poti za uravnavanje procesa invazije. Ciljanje CCL5/CCR5 ni povečalo občutljivosti GBO na ST. V nadaljnjih raziskavah bi uporaba kompleksnejših predkliničnih modelov dodatno pojasnila mehanizme kompenzacijskih signalnih poti ter natančneje opredelila učinek antagonistov in s tem prispevala k večji učinkovitosti celičnih terapij preko krepitve bolnikovega imunskega sistema.

• Jezik: Slovenski jezik
• Ključne besede: glioblastom, glioblastomski organoidi, sferoidi, imunosupresivno tumorsko mikrookolje, standardno zdravljenje, galunisertib, maravirok
• Vrsta gradiva: Magistrsko delo/naloga
• Tipologija: 2.09 - Magistrsko delo
• Organizacija: BF - Biotehniška fakulteta
• Založnik: [E. Pinter]
• Leto izida: 2026
• PID: 20.500.12556/RUL-184741
• UDK: 616.714.1-006:577.2(043.2)
• COBISS.SI-ID: 285037571
• Datum objave v RUL: 15.07.2026

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: The effects of antagonists of tyrosine kinase receptor TGF-βRI and chemokine receptor CCR5 in 3D cell models of glioblastoma

Izvleček

In glioblastoma (GBM), we encounter many challenges, including resistance to radiotherapy and chemotherapy, as well as an immunosuppressive tumor microenvironment (TME). Tumor cells secrete immunosuppressive molecules, such as transforming growth factor beta (TGF-β) and C-C chemokine ligand 5 (CCL5), which recruit and reprogram immune cells to support tumor growth. Novel therapeutic strategies therefore focus on combining different approaches targeting the TME. The aim of this master's thesis was to evaluate the effects of antagonists targeting the serine/threonine kinase receptor TGF-βRI and the chemokine receptor CCR5 on the viability and invasion of three-dimensional (3D) GBM cell models. Furthermore, we investigated whether targeting these cytokine pathways increased the sensitivity of spheroids and glioblastoma organoids (GBOs) to standard therapy (ST). Comparison of the viability and invasion of spheroids derived from GBM patients demonstrated pronounced heterogeneity in response to the TGF-βRI inhibitor, as monotherapy and in combination with ST, between patients and between spheroids generated from differentiated glioblastoma cells (dGBM) and glioblastoma stem cells (GSCs). In two out of six spheroids, inhibition of TGF-βRI with galunisertib (GAL) increased sensitivity to ST. Western blot analysis revealed an impaired SMAD (suppressor of mother against decapentaplegic)-dependent TGF-β pathway, suggesting the presence of a compensatory pathway regulating invasion. Following inhibition with GAL, TGF-βRI expression increased in some spheroids and decreased in others, while inhibition of TGF-β signaling in all dGBM/GSC spheroids activated the SMAD-independent ERK pathway. Maraviroc (MRV), a CCR5 antagonist, had no effect on GBO viability when used as monotherapy. However, it increased GBO invasion, which may be a consequence of activation of a compensatory pathway regulating invasion. Targeting the CCL5/CCR5 axis did not increase GBO sensitivity to ST. In future studies, the use of more complex preclinical models could further elucidate compensatory signaling mechanisms and more precisely define the effects of antagonists, thereby improving the efficacy of cell-based therapies through enhancement of the patient's immune system.

• Ključne besede: glioblastoma, glioblastoma organoids, spheroids, immunosuppressive tumor microenvironment, standard therapy, galunisertib, maraviroc