Jørgen Wesche

• Holme, Børge; Bjørnerud, Birgitte; Pedersen, Nina Marie; Rodriguez de la Ballina, Laura; Wesche, Jørgen & Haugsten, Ellen Margrethe (2023). Automated tracking of cell migration in phase contrast images with CellTraxx. Scientific Reports. ISSN 2045-2322. 13(1). doi: 10.1038/s41598-023-50227-9. Fulltekst i vitenarkiv Vis sammendrag

  The ability of cells to move and migrate is required during development, but also in the adult in processes such as wound healing and immune responses. In addition, cancer cells exploit the cells’ ability to migrate and invade to spread into nearby tissue and eventually metastasize. The majority of cancer deaths are caused by metastasis and the process of cell migration is therefore intensively studied. A common way to study cell migration is to observe cells through an optical microscope and record their movements over time. However, segmenting and tracking moving cells in phase contrast time-lapse video sequences is a challenging task. Several tools to track the velocity of migrating cells have been developed. Unfortunately, most of the automated tools are made for fluorescence images even though unlabelled cells are often preferred to avoid phototoxicity. Consequently, researchers are constrained with laborious manual tracking tools using ImageJ or similar software. We have therefore developed a freely available, user-friendly, automated tracking tool called CellTraxx. This software makes it easy to measure the velocity and directness of migrating cells in phase contrast images. Here, we demonstrate that our tool efficiently recognizes and tracks unlabelled cells of different morphologies and sizes (HeLa, RPE1, MDA-MB-231, HT1080, U2OS, PC-3) in several types of cell migration assays (random migration, wound healing and cells embedded in collagen). We also provide a detailed protocol and download instructions for CellTraxx.

• Nakken, Sigve; Gundersen, Sveinung; Bernal, Fabian Leonardo Martinez; Polychronopoulos, Dimitris; Hovig, Eivind & Wesche, Jørgen (2023). Comprehensive interrogation of gene lists from genome-scale cancer screens with oncoEnrichR. International Journal of Cancer. ISSN 0020-7136. 153(10), s. 1819–1828. doi: 10.1002/ijc.34666. Fulltekst i vitenarkiv
• Fiorito, Elisa; Szybowska, Patrycja Maria; Haugsten, Ellen Margrethe; Kostas, Michal Janusz; Øy, Geir Frode & Wiedlocha, Antoni [Vis alle 12 forfattere av denne artikkelen] (2022). Strategies to inhibit FGFR4 V550L-driven rhabdomyosarcoma. British Journal of Cancer. ISSN 0007-0920. 127, s. 1939–1953. doi: 10.1038/s41416-022-01973-6. Fulltekst i vitenarkiv
• Vikan, Aurora; Haugsten, Ellen Margrethe; Kostas, Michal Janusz; Selbo, Pål Kristian & Wesche, Jørgen (2021). Efficacy and selectivity of fgf2-saporin cytosolically delivered by pci in cells overexpressing fgfr1. Cells. ISSN 2073-4409. 10(6), s. 1–16. doi: 10.3390/cells10061476. Fulltekst i vitenarkiv
• Munson, Michael Joseph; Mathai, Benan J.; Ng, Matthew; Trachsel Moncho, Laura Cristina; de la Ballina, Laura R & Schultz, Sebastian W. [Vis alle 13 forfattere av denne artikkelen] (2021). GAK and PRKCD are positive regulators of PRKN-independent mitophagy. Nature Communications. ISSN 2041-1723. 12(1). doi: 10.1038/s41467-021-26331-7. Fulltekst i vitenarkiv Vis sammendrag

  The mechanisms involved in programmed or damage-induced removal of mitochondria by mitophagy remains elusive. Here, we have screened for regulators of PRKN-independent mitophagy using an siRNA library targeting 197 proteins containing lipid interacting domains. We identify Cyclin G-associated kinase (GAK) and Protein Kinase C Delta (PRKCD) as regulators of PRKN-independent mitophagy, with both being dispensable for PRKN- dependent mitophagy and starvation-induced autophagy. We demonstrate that the kinase activity of both GAK and PRKCD are required for efficient mitophagy in vitro, that PRKCD is present on mitochondria, and that PRKCD facilitates recruitment of ULK1/ATG13 to early autophagic structures. Importantly, we demonstrate in vivo relevance for both kinases in the regulation of basal mitophagy. Knockdown of GAK homologue (gakh-1) in C. elegans or knockout of PRKCD homologues in zebrafish led to significant inhibition of basal mitophagy, highlighting the evolutionary relevance of these kinases in mitophagy regulation.

• Szybowska, Patrycja Maria; Kostas, Michal Janusz; Wesche, Jørgen; Haugsten, Ellen Margrethe & Wiedlocha, Antoni (2021). Negative Regulation of FGFR (Fibroblast Growth Factor Receptor) Signaling. Cells. ISSN 2073-4409. 10(6), s. 1–18. doi: 10.3390/cells10061342. Fulltekst i vitenarkiv
• Tadele, Dagim Shiferaw; Robertson, Joseph; Crispin, Richard; Herrera, Carmen; Chlubnová, Markéta & Piechaczyk, Laure Isabelle [Vis alle 14 forfattere av denne artikkelen] (2020). A cell competition-based small molecule screen identifies a novel compound that induces dual c-Myc depletion and p53 activation. Journal of Biological Chemistry. ISSN 0021-9258. 296. doi: 10.1074/jbc.RA120.015285.
• Szybowska, Patrycja Maria; Kostas, Michal Janusz; Wesche, Jørgen; Wiedlocha, Antoni & Haugsten, Ellen Margrethe (2019). Cancer Mutations in FGFR2 Prevent a Negative Feedback Loop Mediated by the ERK1/2 Pathway. Cells. ISSN 2073-4409. 8(6), s. 1–18. doi: 10.3390/cells8060518. Fulltekst i vitenarkiv
• Zhen, Yan; Haugsten, Ellen Margrethe; Singh, Sachin Kumar & Wesche, Jørgen (2018). Proximity Labeling by a Recombinant APEX2-FGF1 Fusion Protein Reveals Interaction of FGF1 with the Proteoglycans CD44 and CSPG4. Biochemistry. ISSN 0006-2960. 57(26), s. 3807–3816. doi: 10.1021/acs.biochem.8b00120. Vis sammendrag

  Fibroblast growth factor 1 (FGF1) binds to specific FGF receptors (FGFRs) at the surface of target cells to initiate intracellular signaling. While heparan sulfate proteoglycans (HSPGs) are well-described coreceptors, it is uncertain whether there are additional binding sites for FGF1 at the cell surface. To address this, we devised and tested a method to identify novel binding sites for FGF1 at the cell surface, which may also be applicable for other protein ligands. We constructed an APEX2–FGF1 fusion protein to perform proximal biotin labeling of proteins following binding of the fusion protein to the cell surface. After functional validation of the fusion protein by a signaling assay, we used this method to identify binding sites for FGF1 on cell surfaces of living cells. We confirmed the feasibility of our approach by detection of FGFR4, a well-known and specific receptor for FGF1. We subsequently screened for novel interactors using RPE1 cells and identified the proteoglycans CSPG4 (NG2) and CD44. We found that FGF1 binds CD44 through its heparin-binding moiety. Moreover, we found that FGF1 was colocalized with both CSPG4 and CD44 at the cell surface, suggesting that these receptors act as storage molecules that create a reservoir of FGF1. Importantly, our data demonstrate that recombinant ligand–APEX2 fusion proteins can be used to identify novel receptor interactions on the cell surface.

• Fafílek, Bohumil; Bálek, Lukáš; Bosakova, Michaela Kunova; Vařecha, Miroslav; Nita, Alexandru & Gregor, Tomas [Vis alle 20 forfattere av denne artikkelen] (2018). The inositol phosphatase SHIP2 enables sustained ERK activation downstream of FGF receptors by recruiting Src kinases. Science Signaling. ISSN 1945-0877. 11:eaap8608(548), s. 1–15. doi: 10.1126/scisignal.aap8608. Fulltekst i vitenarkiv
• Kostas, Michal Janusz; Haugsten, Ellen Margrethe; Zhen, Yan; Sørensen, Vigdis; Szybowska, Patrycja Maria & Fiorito, Elisa [Vis alle 11 forfattere av denne artikkelen] (2018). Protein Tyrosine Phosphatase Receptor Type G (PTPRG) controls Fibroblast Growth Factor Receptor (FGFR) 1 activity and influences sensitivity to FGFR kinase inhibitors. Molecular & Cellular Proteomics. ISSN 1535-9476. 17(5), s. 850–870. doi: 10.1074/mcp.RA117.000538. Fulltekst i vitenarkiv Vis sammendrag

  Recently, FGFR1 was found to be overexpressed in osteosarcoma and represents an important target for precision medicine. However, because targeted cancer therapy based on FGFR inhibitors has so far been less efficient than expected, a detailed understanding of the target is important. We have here applied proximity-dependent biotin labeling combined with label-free quantitative mass spectrometry to identify determinants of FGFR1 activity in an osteosarcoma cell line. Many known FGFR interactors were identified (e.g. FRS2, PLCG1, RSK2, SRC), but the data also suggested novel determinants. A strong hit in our screen was the tyrosine phosphatase PTPRG. We show that PTPRG and FGFR1 interact and colocalize at the plasma membrane where PTPRG directly dephosphorylates activated FGFR1. We further show that osteosarcoma cell lines depleted for PTPRG display increased FGFR activity and are hypersensitive to stimulation by FGF1. In addition, PTPRG depletion elevated cell growth and negatively affected the efficacy of FGFR kinase inhibitors. Thus, PTPRG may have future clinical relevance by being a predictor of outcome after FGFR inhibitor treatment.

• Haugsten, Ellen Margrethe; Sørensen, Vigdis; Kunova Bosakova, Michaela; De Souza, Gustavo Antonio; Krejci, Pavel & Wiedlocha, Antoni [Vis alle 7 forfattere av denne artikkelen] (2016). Proximity Labeling Reveals Molecular Determinants of FGFR4 Endosomal Transport. Journal of Proteome Research. ISSN 1535-3893. 15(10), s. 3841–3855. doi: 10.1021/acs.jproteome.6b00652.
• Nadratowska-Wesolowska, Beata; Haugsten, Ellen Margrethe; Zakrzewska, Malgorzata; Jakimowicz, Piotr; Zhen, Yan & Pajdzik, Dorota [Vis alle 8 forfattere av denne artikkelen] (2014). RSK2 regulates endocytosis of FGF receptor 1 by phosphorylation on serine 789. Oncogene. ISSN 0950-9232. 33(40), s. 4823–4836. doi: 10.1038/onc.2013.425.
• Oppelt, Angela; Haugsten, Ellen Margrethe; Zech, Tobias; Danielsen, Håvard Emil; Sveen, Anita & Lobert, Viola Helene [Vis alle 8 forfattere av denne artikkelen] (2014). PIKfyve, MTMR3 and their product PtdIns5P regulate cancer cell migration and invasion through activation of Rac1. Biochemical Journal. ISSN 0264-6021. 461(3), s. 383–390. doi: 10.1042/BJ20140132.
• Haugsten, Ellen Margrethe; Brech, Andreas; Liestøl, Knut; Norman, Jim C. & Wesche, Jørgen (2014). Photoactivation Approaches Reveal a Role for Rab11 in FGFR4 Recycling and Signalling. Traffic: The moving front of cell biology. ISSN 1398-9219. 15(6), s. 665–683. doi: 10.1111/tra.12168.
• Haugsten, Ellen Margrethe; Oppelt, Angela & Wesche, Jørgen (2013). Phosphatidylinositol 5-phosphate is a second messenger important for cell migration. Communicative & Integrative Biology. ISSN 1942-0889. 6(5). doi: 10.4161/cib.25446.
• Oppelt, Angela; Lobert, Viola Helene; Haglund, Kaisa; Mackey, Ashley M.; Rameh, Lucia E. & Liestøl, Knut [Vis alle 14 forfattere av denne artikkelen] (2013). Production of phosphatidylinositol 5-phosphate via PIKfyve and MTMR3 regulates cell migration. EMBO Reports. ISSN 1469-221X. 14(1), s. 57–64. doi: 10.1038/embor.2012.183.
• Zakrzewska, Malgorzata; Haugsten, Ellen Margrethe; Nadratowska-Wesolowska, Beata; Oppelt, Angela; Hausott, Barbara & Jin, Yixin [Vis alle 9 forfattere av denne artikkelen] (2013). ERK-Mediated Phosphorylation of Fibroblast Growth Factor Receptor 1 on Ser(777) Inhibits Signaling. Science Signaling. ISSN 1945-0877. 6(262). doi: 10.1126/scisignal.2003087.
• Wesche, Jørgen; Haglund, Kaisa & Haugsten, Ellen Margrethe (2011). Fibroblast growth factors and their receptors in cancer. Biochemical Journal. ISSN 0264-6021. 437, s. 199–213. doi: 10.1042/BJ20101603.
• Haugsten, Ellen Margrethe; Zakrzewska, Malgorzata; Brech, Andreas; Pust, Sascha; Olsnes, Sjur & Sandvig, Kirsten [Vis alle 7 forfattere av denne artikkelen] (2011). Clathrin- and Dynamin-Independent Endocytosis of FGFR3-Implications for Signalling. PLOS ONE. ISSN 1932-6203. 6(7). doi: 10.1371/journal.pone.0021708.
• Haugsten, Ellen Margrethe; Wiedlocha, Antoni; Olsnes, Sjur & Wesche, Jørgen (2010). Roles of Fibroblast Growth Factor Receptors in Carcinogenesis. Molecular Cancer Research. ISSN 1541-7786. 8(11), s. 1439–1452. doi: 10.1158/1541-7786.MCR-10-0168.
• Haglund, Kaisa; Nezis, Jonnis P; Lemus Torres, Dafne; Grabbe, C; Wesche, Jørgen & Liestøl, Knut [Vis alle 9 forfattere av denne artikkelen] (2010). Cindr Interacts with Anillin to Control Cytokinesis in Drosophila melanogaster. Current Biology. ISSN 0960-9822. 20(10), s. 944–950. doi: 10.1016/j.cub.2010.03.068.
• Lobert, Viola Helene; Brech, Andreas; Pedersen, Nina Marie; Wesche, Jørgen; Oppelt, Angela & Malerød, Lene [Vis alle 7 forfattere av denne artikkelen] (2010). Ubiquitination of alpha 5 beta 1 lntegrin Controls Fibroblast Migration through Lysosomal Degradation of Fibronectin-lntegrin Complexes. Developmental Cell. ISSN 1534-5807. 19(1), s. 148–159. doi: 10.1016/j.devcel.2010.06.010.
• Zakrzewska, Malgorzata; Zhen, Yan; Wiedlocha, Antoni; Olsnes, Sjur & Wesche, Jørgen (2009). Size Limitation in Translocation of Fibroblast Growth Factor 1 Fusion Proteins across the Endosomal Membrane. Biochemistry. ISSN 0006-2960. 48(30), s. 7209–7218. doi: 10.1021/bi9007353.
• Haugsten, Ellen Margrethe; Malecki, Jedrzej M.; Bjørklund, Sunniva Maria Stordal; Olsnes, Sjur & Wesche, Jørgen (2008). Ubiquitination of fibroblast growth factor receptor 1 is required for its intracellular sorting but not for its endocytosis. Molecular Biology of the Cell. ISSN 1059-1524. 19, s. 3390–3403. doi: 10.1091/mbc.E07-12-1219.
• Nilsen, Trine; Rosendal, Ken; Sørensen, Vigdis; Wesche, Jørgen; Olsnes, Sjur & Widocha, Antoni (2007). A nuclear export sequence located on a beta-strand in fibroblast growth factor-1. Journal of Biological Chemistry. ISSN 0021-9258. 282(36), s. 26245–26256. doi: 10.1074/jbc.M611234200.
• Wesche, Jørgen; Malecki, Jedrzej M.; Wiedlocha, Antoni; Skjerpen, Camilla Skiple; Claus, P & Olsnes, Sjur (2006). FGF-1 and FGF-2 require the cytosolic chaperone Hsp90 for translocation into the cytosol and the cell nucleus. Journal of Biological Chemistry. ISSN 0021-9258. 281, s. 11405–11412.
• Sørensen, Vigdis; Wiedlocha, Antoni; Haugsten, Ellen Margrethe; Khnykin, Denis; Wesche, Jørgen & Olsnes, Sjur (2006). Different abilities of the four FGFRs to mediate FGF-1 translocation are linked to differences in the receptor C-terminal tail. Journal of Cell Science. ISSN 0021-9533. 119, s. 4332–4341.
• Wiedlocha, Antoni; Nilsen, Trine; Wesche, Jørgen; Sørensen, Vigdis; Malecki, M & Marcinkowska, Ewa [Vis alle 7 forfattere av denne artikkelen] (2005). Phosphorylation-regulated nucleocytoplasmic trafficking of internalized fibroblast growth factor-1. Molecular Biology of the Cell. ISSN 1059-1524. s. 794–810.
• Haugsten, Ellen Margrethe; Sørensen, Vigdis; Olsnes, Sjur; Wesche, Jørgen & Brech, Andreas (2005). Different intracellular trafficking of FGF1 endocytosed by the four specific and homologous FGF receptors. Journal of Cell Science. ISSN 0021-9533. 118, s. 3869–3881.
• Wesche, Jørgen; Malecki, Jedrzej M.; Wiedlocha, Antoni; Ehsani, Maryam; Nilsen, Trine & Marcinkowska, Ewa [Vis alle 7 forfattere av denne artikkelen] (2005). Two nuclear localization signals required for transport from the cytosol to the, nucleus of externally added FGF-1 translocated into cells. Biochemistry (Moscow). ISSN 0006-2979. 44.
• Malecki, Jedrzej; Wesche, Jørgen; Skjerpen, Camilla Skiple; Wiedlocha, Antoni & Olsnes, Sjur (2004). Translocation of FGF-1 and FGF-2 across Vesicular Membranes Occurs during G1-Phase by a Common Mechanism. Molecular Biology of the Cell. ISSN 1059-1524. 15, s. 804–814.

Se alle arbeider i Cristin

• Wesche, Jørgen (2022). Activation of FGFR1 by inorganic phosphate and oxidative stress.
• Wesche, Jørgen (2022). Jakten på molekylene som hjelper kreftceller å spre seg til nye steder i kroppen.
• Skjerpen, Camilla Skiple; Olsnes, Sjur & Wesche, Jørgen (2005). Translocation mechanism and cellular interaction partners of FGF-1 and FGF-2. Unipub forlag. ISSN 82-8072-158-4.

Se alle arbeider i Cristin