The Hedgehog signaling pathway has been implicated in development of various tumors, but the exact roles and molecular mechanisms are still not understood. The focus of this research are the tumors harboring BRAF or NRAS mutations, and their differential response to Hedgehog pathway inhibition. Both BRAF and NRAS non-canonically activate the three GLI proteins, but the activity and interplay of this interaction is still unclear. With this project we propose to determine the exact transcriptional targets of GLI1, GLI2 and GLI3 proteins in melanoma with different genetic backgrounds, either harboring a BRAF mutation, a NRAS mutation, or no mutation in these two genes. For this purpose, we will first perform ChIP-seq to determine the transcriptional targets, then knock-out each of the three GLI proteins and sequence the transcriptomes of the knock-out cell lines. After the knock-outs have been generated, a series of cell assays will be performed to determine the role of each of the three GLI proteins in BRAF/NRAS dependent tumors, and to elucidate the mechanism of their action in melanoma. Combined treatments with Hedgehog inhibitors and specific BRAF/NRAS inhibitors, followed by generation and testing of resistant cell lines, will demonstrate if there is a potential benefit in the clinical management of melanoma. Finally, in the final year of the project, a set of clinical samples will be analyzed for these potential biomarkers to determine their possible use in the clinic. In summary, this project plans a complete analysis of the GLI code in BRAF/NRAS dependent melanoma, from determining the transcriptional targets and finding molecular mechanisms, to potential translation to the clinic.

Journal articles

1. Kurtović, Matea; Piteša, Nikolina; Bartoniček, Nenad; Ozretić, Petar; Musani, Vesna; Čonkaš, Josipa; Petrić, Tina; King, Cecile; Sabol, Maja
RNA-seq and ChIP-seq Identification of Unique and Overlapping Targets of GLI Transcription Factors in Melanoma Cell Lines.  // Cancers, 14 (2022), 18; 4540, 20 doi:10.3390/cancers14184540 (international peer review, article, scholarly)
2. Trnski, Diana; Sabol, Maja; Tomić, Sanja; Štefanac, Ivan; Mrčela, Milanka; Musani, Vesna; Rinčić, Nikolina; Kurtović, Matea; Petrić, Tina; Levanat, Sonja; Ozretić, Petar
SHH-N non-canonically sustains androgen receptor activity in androgen-independent prostate cancer cells.  // Scientific reports, 11 (2021), 1; 14880, 11 doi:10.1038/s41598-021-93971-6 (international peer review, article, scholarly)
3. Zubčić*, Vedran; Rinčić*, Nikolina; Kurtović, Matea; Trnski, Diana; Musani, Vesna; Ozretić, Petar; Levanat, Sonja; Leović, Dinko; Sabol, Maja
GANT61 and Lithium Chloride Inhibit the Growth of Head and Neck Cancer Cell Lines Through the Regulation of GLI3 Processing by GSK3beta.  // International journal of molecular sciences, 21 (2020), 6410, 13 doi:10.3390/ijms21176410 (international peer review, article, scholarly)

Conference abstracts

1. Čonkaš, Josipa; Josić, Janja; Piteša, Nikolina; Kurtović, Matea; Petrić, Tina; Musani, Vesna; Sabol, Maja; Vugrinec, Ozren; Leović, Dinko; Ozretić, Petar
The role of androgen and androgen receptors in metastasis of head and neck tumors.  // 6. simpozij studenata doktorskih studija PMF-a : knjiga sažetaka = 6th Faculty of Science PhD student symposium : book of abstracts / Schneider, Petra (ed.).
Zagreb: Prirodoslovno-matematički fakultet, Sveučilište u Zagrebu, 2022. pp. 192-193 (poster, local peer review, abstract, scholarly)
2. Kurtović, Matea; Piteša, Nikolina; Bartoniček, Nenad; Ozretić, Petar; Musani, Vesna; Petrić, Tina; Čonkaš, Josipa; Sabol, Maja
Unique and overlapping transcriptional targets of GLI1, GLI2 and GLI3 in melanoma cell lines.  // “HDIR-6: Targeting Cancer” The 6th Meeting of the Croatian Association for Cancer Research with International Participation / Ozretić, Petar - Zagreb : Croatian Association for Cancer Research, 2022
Zagreb, Hrvatska, 2022. pp. 25-25 (lecture, international peer review, abstract, scholarly)
3. Petrić, Tina; Kurtović, Matea; Piteša, Nikolina; Čonkaš, Josipa; Musani, Vesna; Ozretić, Petar; Sabol, Maja
3D CULTURE OF PROSTATE CANCER AS A MODEL TO STUDY HEDGEHOG-GLI SIGNALING PATHWAY.  // “HDIR-6: Targeting Cancer” The 6th Meeting of the Croatian Association for Cancer Research with International Participation / Ozretić, Petar (ed.).
Zagreb: Croatian Association for Cancer Research, 2022. pp. 44-44 (poster, abstract, scholarly)
4. Petrić, Tina; Kurtović, Matea; Piteša, Nikolina; Čonkaš, Josipa; Musani, Vesna; Ozretić, Petar; Sabol, Maja
Hedgehog-GLI signaling pathway in 3D model of prostate cancer.  // From Science to Knowledge / Dulić, Morana ; Sinčić, Nino ; Vrhovac Madunić, Ivana (ed.).
Zagreb: HDBMB, 2022. pp. 117-117 (poster, abstract, scholarly)
5. Piteša, Nikolina; Bartoniček, Nenad; Kurtović, Matea; Petrić, Tina; Čonkaš, Josipa; Musani, Vesna; Ozretić, Petar; Sabol, Maja
Signaling Switch from Hedgehog-GLI to MAPK Potentially Drives Primary Cilia Loss inNRAS Mutated GANT61-Resistant Melanoma Cell Line.  // “HDIR-6: Targeting Cancer” - The 6th Meeting of the Croatian Association for Cancer Research with International Participation – Book of Abstracts / Ozretić, Petar (ed.).
Zagreb: Croatian Association for Cancer Research, Zagreb, Croatia, 2022. pp. 45-45 (poster, international peer review, abstract, scholarly)
6. Čonkaš, Josipa; Josić, Janja; Piteša, Nikolina; Kurtović, Matea; Petrić, Tina; Musani, Vesna; Sabol, Maja; Vugrinec, Ozren; Leović, Dinko; Ozretić, Petar
Membrane androgen receptor OXER1 indicates apotential role in metastasis of head and neck tumors.  // “HDIR-6: Targeting Cancer” - The 6th Meeting of the Croatian Association for Cancer Research with International Participation – Book of Abstracts / Ozretić, Petar (ed.).
Zagreb: Croatian Association for Cancer Research, 2022. pp. 36-36 (poster, international peer review, abstract, scholarly)
7. Piteša, Nikolina; Bartoniček, Nenad; Kurtović, Matea; Petrić, Tina; Čonkaš, Josipa; Musani, Vesna; Ozretić, Petar; Sabol, Maja
Primary cilia - a potential link between hedgehog- GLI and MAPK signaling in GANT61 -resistant  melanoma cell lines.  // Book of Abstracts of the Congress of the Croatian Society of Biochemistry and Molecular Biology HDBMB22: From Science to Knowledge / Dulić, Morana ; Sinčić, Nino ; Vrhovac Madunić Ivana (ed.).
Zagreb: Croatian Society of Biochemistry and Molecular Biology, 2022. pp. 119-119 (poster, international peer review, abstract, scholarly)
8. Piteša, N; Kurtović, M; Bartoniček, N; Petrić, T; Čonkaš, J; Musani, V; Ozretić, P; Sabol, M
Hedgehog-GLI and MAPK signalling pathway activity in GANT61-resistant melanoma cells.  // Online abstracts
Seville, Španjolska, 2022. EACR22-0296, 1 (poster, information about peer review not available, abstract, other)
9. Kurtović, Matea; Rinčić, Nikolina; Trnski, Diana; Musani, Vesna; Ozretić, Petar; Sabol, Maja
CRISPR/Cas9 mediated knockout of GLI1, GLI2 and GLI3 genes in melanoma cell lines.  // 45th FEBS Congress Abstracts
Virtualno, 2021. 38528, 1 (poster, abstract, scholarly)
10. Piteša, Nikolina; Bartoniček, Nenad; Kurtović, Matea; Musani, Vesna; Trnski, Diana; Ozretić, Petar; Sabol, Maja
Genes for competing endogenous RNAs as targets of transcription factors GLI in melanoma cell lines.  // Serbian Association for Cancer Research-5th Congress of SDIR: Translational Potential of Cancer Research in Serbia (Abstract book) / Đorđić Crnogorac, Marija ; Nedeljković, Milica (ed.).
Beograd: Srpsko društvo istraživača raka, 2021. pp. 9-9 (lecture, international peer review, abstract, scholarly)
11. Križanac, Marinela; Leović, Matko; Rinčić, Nikolina; Trnski, Diana; Musani, Vesna; Ozretić, Petar; Levanat, Sonja; Zubčić, Vedran; Leović, Dinko; Sabol, Maja
Hedgehog-GLI signaling controls proliferation and invasiveness of head and neck squamous cell carcinoma.  // HDBMB2019 Crossroads in Life Sciences / Katalinić, Maja ; Dulić, Morana ; Stuparević, Igor (ed.).
Zagreb: Croatian Society of Biochemistry and Molecular Biology, 2019. pp. 115-115 (poster, information about peer review not available, abstract, scholarly)

Other conference submissions

1. Kurtović, Matea; Bartoniček, Nenad; Piteša, Nikolina; Ozretić, Petar; Musani, Vesna; Petrić, Tina; Čonkaš, Josipa; Sabol, Maja
RNA sequencing reveals unique and overlapping transcriptional targets of GLI1, GLI2 and GLI3 in melanoma cell lines.  // EACR 2022 Congress - Innovative Cancer Science: Translating Biology to Medicine
Sevilla, Španjolska, 2022. (poster, other, scholarly)
2. Petrić, Tina
Let's Go 3D: The Future Of Prostate Cancer Models.  // THE KEMOMIND SCIENCE CONFERENCE / Kokondoska Grgič, Vesna ; Vuga, Andrej (ed.).
Ljubljana: Kemomed d.o.o., 2022. pp. 32-32 (lecture, abstract, scholarly)
3. Rinčić, Nikolina; Bartoniček, Nenad; Kurtović, Matea; Musani, Vesna; Trnski, Diana; Ozretić, Petar; Sabol, Maja
Specific and overlapping GLI transcription targets in melanoma.  // EACR 2021 : : Innovative Cancer Science
virtualni kongres, 2021. (poster, information about peer review not available, unpublished, scholarly)
4. Kurtović, Matea; Rinčić, Nikolina; Trnski, Diana; Musani, Vesna; Ozretić, Petar; Sabol, Maja
CRISPR/Cas9 mediated knockout of GLI1, GLI2 and GLI3 genes in melanoma cell lines.  // 20th FEBS Young Scientists´Forum
Virtualno, 2021. 38528, 1 (poster, abstract, scholarly)
5. Kurtović, Matea; Rinčić, Nikolina; , Trnski, Diana; Musani, Vesna; Ozretić, Petar; Sabol, Maja
Generation of GLI1, GLI2 and GLI3 knock-out melanoma cell lines via CRISPR/Cas9 gene editing.  // EACR 2021 Virtual Congress
Virtual, 2021. EACR21v 0165, 1 (poster, abstract, scholarly)
6. Petrić, Tina; Trnski, Diana; Kurtović, Matea; Rinčić, Nikolina; Musani, Vesna; Ozretić, Petar; Sabol, Maja
Hedgehog-GLI signaling pathway in 3D model of prostate cancer.  // Goodbye Flat Biology: Next Generation Cancer Models
Virtual event, 2021. (poster, international peer review, unpublished, scholarly)
7. Rinčić, Nikolina; Nenad, Bartoniček; Matea, Kurtović; Vesna, Musani; Diana, Trnski; Petar, Ozretić; Maja, Sabol
GLI transcription targets in melanoma cell lines.  // 45th FEBS Virtual Congress “Molecules of Life: Towards New Horizons“
virtualni kongres, 2021. (poster, peer review, unpublished, scholarly)

Theses

1. Piteša, Nikolina. Interaction of RAS/RAF/MAPK and Hedgehog-GLI signaling pathways in therapy resistance in human melanoma cell lines., 2023, Doctoral dissertation, JJ Strossmayer University of Osijek, Osijek (Mentor: Maja Sabol)
2. Vučemilo, Nikolina. Establishing a three-dimensional spheroid culture of head and neck tumor cells., 2022, Diploma thesis, Department of Biology, Faculty of Science, University of Zagreb, Zagreb (Mentor: Maja Sabol)
3. Šimatović, Domagoj. Role of p16 expression in head and neck carcinoma., 2021, Diploma thesis, Department of Biology, Faculty of Science, University of Zagreb, Zagreb (Mentor: Vesna Musani)
4. Brtan, Ana. Characterization and validation of SHH knock-in melanoma cell lines., 2021, Diploma thesis, Department of Biology, Faculty of Science, University of Zagreb, Zagreb (Mentor: Maja Sabol)
5. Čonkaš, Josipa. Construction of vectors for SHH protein expression in CHL-1 human melanoma cell line., 2021, Diploma thesis, Department of Biology, Faculty of Science, University of Zagreb, Zagreb (Mentor: Maja Sabol)
6. Rinčić, Nikolina. CRISPR/Cas9-mediated generation of GLI1 knock-out cell line., 2018, Diploma thesis, Department of Biology, Faculty of Science, University of Zagreb, Zagreb (Mentor: Maja Sabol)

Prizes and Awards

1. Čonkaš, Josipa: Annual award of the Croatian Association of Genetic Engineers "Zoran Zgaga" for the best graduate thesis in which genetic engineering methods were applied, defended in the academic year 2020/2021
2. Rinčić, Nikolina: The Annual Award of the Ruđer Bošković Institute for the scientific paper published in 2020, entitled "GANT61 and Lithium Chloride Inhibit the Growth of Head and Neck Cancer Cell Lines Through the Regulation of GLI3 Processing by GSK3beta"