Naslov
Characterization of GANT61 Resistant Cell Lines reveals new insights of Hedgehog-GLI and RAS/RAF/MAPK interplay in melanoma

Autori
Piteša, Nikolina ; Bartoniček, Nenad ; Kurtović, Matea ; Petrić, Tina ; Čonkaš, Josipa ; Musani, Vesna ; Ozretić, Petar ; Sabol, Maja

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
EACR-Boehringer Ingelheim Drugging and Regulating the MAP Kinase Pathway

Mjesto i datum
Online event, 21.02.2023. - 22.02.2023

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
melanoma ; HH-GLI ; RAS/RAF/MAPK ; primary cilia ; therapy resistance

Sažetak
Hedgehog-GLI (HH-GLI) is an important embryonal signaling pathway often deregulated in melanoma. Beside its canonical activity, which is highly dependent on the primary cilia, it can be activated noncanonically through other signaling pathways like RAS/RAF/MAPK. Although some mechanisms have been previously described, mediators of these interactions and final outcomes are still poorly understood. Better understanding of these interaction may be crucial in establishment and maintenance of drug resistance, a known issue for metastatic melanoma treatment. To investigate in more depth these interactions, we used two experimental approaches, first based on the identification of GLI transcriptional targets connected to RAS/RAF/MAPK and second, the establishment of resistant cell lines. We successfully established two melanoma cell lines with different NRAS mutation background resistant to GANT61, a specific GLI protein inhibitor. The characterization followed after cell line validation showed that by gaining GANT61 resistance, cell lines changed their response to HH-GLI and RAS/RAF/MAPK inhibitors, migration and colony formation capacity. Beside these changes, established cell lines also modulated HH-GLI and RAS/RAF/MAPK signaling, autophagy activity and cell cycle. Our previous study (Kurtović et al. 2022), using a combined ChIP-seq and RNA-seq approach identified novel GLI transcription targets involved in RAS/RAF/MAPK signaling, therefore we examined if any of these potential target genes are changed in established cell lines. A newly identified GLI2 transcription target and potential MAPK substrate, RAB34 essential for ciliogenesis, was downregulated in NRAS mutated resistant cell line which after all showed also primary cilia loss. Our results suggest that although established cell lines share certain molecular features, they potentially depend on different MAPK pathways. We believe that primary cilia represent a potential link between HH-GLI and MAPK signaling in GANT61 resistant NRAS mutated melanoma. Therefore, our future studies will focus on ciliogenesis regulation via RAS/RAF/MAPK signaling and its function in drug resistance.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Temeljne medicinske znanosti

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