A Novel Role for NUAK1 in Promoting Ovarian Cancer Metastasis through Regulation of Fibronectin Production in Spheroids
Epithelial ovarian cancer (EOC) has a unique mode of metastasis, where cells shed from the primary tumour, form aggregates called spheroids to evade anoikis, spread through the peritoneal cavity, and adhere to secondary sites. We previously showed that the master kinase Liver kinase B1 (LKB1) is req...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-05-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/12/5/1250 |
| _version_ | 1797567918317240320 |
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| author | Jamie Lee Fritz Olga Collins Parima Saxena Adrian Buensuceso Yudith Ramos Valdes Kyle E. Francis Kevin R. Brown Brett Larsen Karen Colwill Anne-Claude Gingras Robert Rottapel Trevor G. Shepherd |
| author_facet | Jamie Lee Fritz Olga Collins Parima Saxena Adrian Buensuceso Yudith Ramos Valdes Kyle E. Francis Kevin R. Brown Brett Larsen Karen Colwill Anne-Claude Gingras Robert Rottapel Trevor G. Shepherd |
| author_sort | Jamie Lee Fritz |
| collection | DOAJ |
| description | Epithelial ovarian cancer (EOC) has a unique mode of metastasis, where cells shed from the primary tumour, form aggregates called spheroids to evade anoikis, spread through the peritoneal cavity, and adhere to secondary sites. We previously showed that the master kinase Liver kinase B1 (LKB1) is required for EOC spheroid viability and metastasis. We have identified novel (nua) kinase 1 (NUAK1) as a top candidate LKB1 substrate in EOC cells and spheroids using a multiplex inhibitor beads-mass spectrometry approach. We confirmed that LKB1 maintains NUAK1 phosphorylation and promotes its stabilization. We next investigated NUAK1 function in EOC cells. Ectopic NUAK1-overexpressing EOC cell lines had increased adhesion, whereas the reverse was seen in OVCAR8-<i>NUAK1</i>KO cells. In fact, cells with NUAK1 loss generate spheroids with reduced integrity, leading to increased cell death after long-term culture. Following transcriptome analysis, we identified reduced enrichment for cell interaction gene expression pathways in OVCAR8-<i>NUAK1</i>KO spheroids. In fact, the <i>FN1</i> gene, encoding fibronectin, exhibited a 745-fold decreased expression in <i>NUAK1</i>KO spheroids. Fibronectin expression was induced during native spheroid formation, yet this was completely lost in <i>NUAK1</i>KO spheroids. Co-incubation with soluble fibronectin restored the compact spheroid phenotype to OVCAR8-<i>NUAK1</i>KO cells. In a xenograft model of intraperitoneal metastasis, NUAK1 loss extended survival and reduced fibronectin expression in tumours. Thus, we have identified a new mechanism controlling EOC metastasis, through which LKB1-NUAK1 activity promotes spheroid formation and secondary tumours via fibronectin production. |
| first_indexed | 2024-03-10T19:48:58Z |
| format | Article |
| id | doaj.art-ca28bd71215d4173b440653d15c3393e |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-10T19:48:58Z |
| publishDate | 2020-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-ca28bd71215d4173b440653d15c3393e2023-11-20T00:37:08ZengMDPI AGCancers2072-66942020-05-01125125010.3390/cancers12051250A Novel Role for NUAK1 in Promoting Ovarian Cancer Metastasis through Regulation of Fibronectin Production in SpheroidsJamie Lee Fritz0Olga Collins1Parima Saxena2Adrian Buensuceso3Yudith Ramos Valdes4Kyle E. Francis5Kevin R. Brown6Brett Larsen7Karen Colwill8Anne-Claude Gingras9Robert Rottapel10Trevor G. Shepherd11The Mary & John Knight Translational Ovarian Cancer Research Unit, London Regional Cancer Program, London, ON N6A 4L6, CanadaThe Mary & John Knight Translational Ovarian Cancer Research Unit, London Regional Cancer Program, London, ON N6A 4L6, CanadaThe Mary & John Knight Translational Ovarian Cancer Research Unit, London Regional Cancer Program, London, ON N6A 4L6, CanadaThe Mary & John Knight Translational Ovarian Cancer Research Unit, London Regional Cancer Program, London, ON N6A 4L6, CanadaThe Mary & John Knight Translational Ovarian Cancer Research Unit, London Regional Cancer Program, London, ON N6A 4L6, CanadaPrincess Margaret Cancer Centre, Toronto, ON M5G 2C1, CanadaTerrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON M5S 3E1, CanadaLunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON M5G 1X5, CanadaLunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON M5G 1X5, CanadaLunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON M5G 1X5, CanadaPrincess Margaret Cancer Centre, Toronto, ON M5G 2C1, CanadaThe Mary & John Knight Translational Ovarian Cancer Research Unit, London Regional Cancer Program, London, ON N6A 4L6, CanadaEpithelial ovarian cancer (EOC) has a unique mode of metastasis, where cells shed from the primary tumour, form aggregates called spheroids to evade anoikis, spread through the peritoneal cavity, and adhere to secondary sites. We previously showed that the master kinase Liver kinase B1 (LKB1) is required for EOC spheroid viability and metastasis. We have identified novel (nua) kinase 1 (NUAK1) as a top candidate LKB1 substrate in EOC cells and spheroids using a multiplex inhibitor beads-mass spectrometry approach. We confirmed that LKB1 maintains NUAK1 phosphorylation and promotes its stabilization. We next investigated NUAK1 function in EOC cells. Ectopic NUAK1-overexpressing EOC cell lines had increased adhesion, whereas the reverse was seen in OVCAR8-<i>NUAK1</i>KO cells. In fact, cells with NUAK1 loss generate spheroids with reduced integrity, leading to increased cell death after long-term culture. Following transcriptome analysis, we identified reduced enrichment for cell interaction gene expression pathways in OVCAR8-<i>NUAK1</i>KO spheroids. In fact, the <i>FN1</i> gene, encoding fibronectin, exhibited a 745-fold decreased expression in <i>NUAK1</i>KO spheroids. Fibronectin expression was induced during native spheroid formation, yet this was completely lost in <i>NUAK1</i>KO spheroids. Co-incubation with soluble fibronectin restored the compact spheroid phenotype to OVCAR8-<i>NUAK1</i>KO cells. In a xenograft model of intraperitoneal metastasis, NUAK1 loss extended survival and reduced fibronectin expression in tumours. Thus, we have identified a new mechanism controlling EOC metastasis, through which LKB1-NUAK1 activity promotes spheroid formation and secondary tumours via fibronectin production.https://www.mdpi.com/2072-6694/12/5/1250ovarian cancerspheroidLKB1NUAK1metastasisfibronectin |
| spellingShingle | Jamie Lee Fritz Olga Collins Parima Saxena Adrian Buensuceso Yudith Ramos Valdes Kyle E. Francis Kevin R. Brown Brett Larsen Karen Colwill Anne-Claude Gingras Robert Rottapel Trevor G. Shepherd A Novel Role for NUAK1 in Promoting Ovarian Cancer Metastasis through Regulation of Fibronectin Production in Spheroids Cancers ovarian cancer spheroid LKB1 NUAK1 metastasis fibronectin |
| title | A Novel Role for NUAK1 in Promoting Ovarian Cancer Metastasis through Regulation of Fibronectin Production in Spheroids |
| title_full | A Novel Role for NUAK1 in Promoting Ovarian Cancer Metastasis through Regulation of Fibronectin Production in Spheroids |
| title_fullStr | A Novel Role for NUAK1 in Promoting Ovarian Cancer Metastasis through Regulation of Fibronectin Production in Spheroids |
| title_full_unstemmed | A Novel Role for NUAK1 in Promoting Ovarian Cancer Metastasis through Regulation of Fibronectin Production in Spheroids |
| title_short | A Novel Role for NUAK1 in Promoting Ovarian Cancer Metastasis through Regulation of Fibronectin Production in Spheroids |
| title_sort | novel role for nuak1 in promoting ovarian cancer metastasis through regulation of fibronectin production in spheroids |
| topic | ovarian cancer spheroid LKB1 NUAK1 metastasis fibronectin |
| url | https://www.mdpi.com/2072-6694/12/5/1250 |
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