Kinome Profiling to Predict Sensitivity to MAPK Inhibition in Melanoma and to Provide New Insights into Intrinsic and Acquired Mechanism of Resistance <break/>Short Title: Sensitivity Prediction to MAPK Inhibitors in Melanoma
Mitogen-activated protein kinase (MAPK) inhibition with the combination of BRAF (Rapidly Accelerated Fibrosarcoma) and MEK (Mitogen-activated protein kinase kinase) inhibitors has become the standard of first-line therapy of metastatic melanoma harbouring BRAF V600 mutations. However, about half of...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-02-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/12/2/512 |
| _version_ | 1797709450285416448 |
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| author | Mohamad Krayem Philippe Aftimos Ahmad Najem Tim van den Hooven Adriënne van den Berg Liesbeth Hovestad-Bijl Rik de Wijn Riet Hilhorst Rob Ruijtenbeek Malak Sabbah Joseph Kerger Ahmad Awada Fabrice Journe Ghanem E. Ghanem |
| author_facet | Mohamad Krayem Philippe Aftimos Ahmad Najem Tim van den Hooven Adriënne van den Berg Liesbeth Hovestad-Bijl Rik de Wijn Riet Hilhorst Rob Ruijtenbeek Malak Sabbah Joseph Kerger Ahmad Awada Fabrice Journe Ghanem E. Ghanem |
| author_sort | Mohamad Krayem |
| collection | DOAJ |
| description | Mitogen-activated protein kinase (MAPK) inhibition with the combination of BRAF (Rapidly Accelerated Fibrosarcoma) and MEK (Mitogen-activated protein kinase kinase) inhibitors has become the standard of first-line therapy of metastatic melanoma harbouring BRAF V600 mutations. However, about half of the patients present with primary resistance while the remaining develop secondary resistance under prolonged treatment. Thus, there is a need for predictive biomarkers for sensitivity and/or resistance to further refine the patient population likely to benefit from MAPK inhibitors. In this study, we explored a top-down approach using a multiplex kinase assay, first, to discover a kinome signature predicting sensitivity, intrinsic and acquired resistance to MAPK inhibitors in melanoma, and second, to understand the mechanism of resistance using cell lines. Pre-dose tissues from patients (four responders and three non-responders to BRAFi monotherapy) were profiled for phosphotyrosine kinase (PTK) and serine-threonine kinase (STK) activities on a PamChip<sup>®</sup> peptide microarray in the presence and absence of ex vivo BRAFi. In addition, molecular studies were conducted on four sensitive parental lines, their offspring with acquired resistance to BRAFi and two lines with intrinsic resistance. PTK and STK activities in cell lysates were measured in the presence and absence of ex vivo BRAFi and/or MEKi. In tissue lysates, concentration-dependent <i>ex vivo</i> inhibition of STK and PTK activities with dabrafenib was stronger in responders than in non-responders. This difference was confirmed in cell lines comparing sensitive and resistant ones. Interestingly, common features of resistance were increased activity of receptor tyrosine kinases, Proto-oncogene tyrosine-protein kinase Src (Src) family kinases and protein kinase B (PKB, AKT) signalling. These latter results were confirmed by Western blots. While dabrafenib alone showed an inhibition of STK and PTK activities in both tissues and cell lines, the combination of dabrafenib and trametinib showed an antagonism on the STK activities and a synergism on PTK activities, resulting in stronger inhibitions of overall tyrosine kinase activities. Altogether; these data reveal that resistance of tumours and cell lines to MAPK inhibitors can be predicted using a multiplex kinase assay and is associated with an increase in specific tyrosine kinase activities and globally to AKT signalling in the patient’s tissue. Thus, such a predictive kinome signature would help to identify patients with innate resistance to MAPK double inhibition in order to propose other therapies. |
| first_indexed | 2024-03-12T06:37:45Z |
| format | Article |
| id | doaj.art-03e5f2913507482d86d31eade456925c |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-12T06:37:45Z |
| publishDate | 2020-02-01 |
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| record_format | Article |
| series | Cancers |
| spelling | doaj.art-03e5f2913507482d86d31eade456925c2023-09-03T01:10:56ZengMDPI AGCancers2072-66942020-02-0112251210.3390/cancers12020512cancers12020512Kinome Profiling to Predict Sensitivity to MAPK Inhibition in Melanoma and to Provide New Insights into Intrinsic and Acquired Mechanism of Resistance <break/>Short Title: Sensitivity Prediction to MAPK Inhibitors in MelanomaMohamad Krayem0Philippe Aftimos1Ahmad Najem2Tim van den Hooven3Adriënne van den Berg4Liesbeth Hovestad-Bijl5Rik de Wijn6Riet Hilhorst7Rob Ruijtenbeek8Malak Sabbah9Joseph Kerger10Ahmad Awada11Fabrice Journe12Ghanem E. Ghanem13Laboratory of Oncology and Experimental Surgery, Institut Jules Bordet, Université Libre de Bruxelles, 1000 Brussels, BelgiumMedical Oncology Clinic, Institut Jules Bordet, Université Libre de Bruxelles, 1000 Brussels, BelgiumLaboratory of Oncology and Experimental Surgery, Institut Jules Bordet, Université Libre de Bruxelles, 1000 Brussels, BelgiumPamGene International BV, 5211HH ’s-Hertogenbosch, The NetherlandsPamGene International BV, 5211HH ’s-Hertogenbosch, The NetherlandsPamGene International BV, 5211HH ’s-Hertogenbosch, The NetherlandsPamGene International BV, 5211HH ’s-Hertogenbosch, The NetherlandsPamGene International BV, 5211HH ’s-Hertogenbosch, The NetherlandsPamGene International BV, 5211HH ’s-Hertogenbosch, The NetherlandsLaboratory of Oncology and Experimental Surgery, Institut Jules Bordet, Université Libre de Bruxelles, 1000 Brussels, BelgiumMedical Oncology Clinic, Institut Jules Bordet, Université Libre de Bruxelles, 1000 Brussels, BelgiumLaboratory of Oncology and Experimental Surgery, Institut Jules Bordet, Université Libre de Bruxelles, 1000 Brussels, BelgiumLaboratory of Oncology and Experimental Surgery, Institut Jules Bordet, Université Libre de Bruxelles, 1000 Brussels, BelgiumLaboratory of Oncology and Experimental Surgery, Institut Jules Bordet, Université Libre de Bruxelles, 1000 Brussels, BelgiumMitogen-activated protein kinase (MAPK) inhibition with the combination of BRAF (Rapidly Accelerated Fibrosarcoma) and MEK (Mitogen-activated protein kinase kinase) inhibitors has become the standard of first-line therapy of metastatic melanoma harbouring BRAF V600 mutations. However, about half of the patients present with primary resistance while the remaining develop secondary resistance under prolonged treatment. Thus, there is a need for predictive biomarkers for sensitivity and/or resistance to further refine the patient population likely to benefit from MAPK inhibitors. In this study, we explored a top-down approach using a multiplex kinase assay, first, to discover a kinome signature predicting sensitivity, intrinsic and acquired resistance to MAPK inhibitors in melanoma, and second, to understand the mechanism of resistance using cell lines. Pre-dose tissues from patients (four responders and three non-responders to BRAFi monotherapy) were profiled for phosphotyrosine kinase (PTK) and serine-threonine kinase (STK) activities on a PamChip<sup>®</sup> peptide microarray in the presence and absence of ex vivo BRAFi. In addition, molecular studies were conducted on four sensitive parental lines, their offspring with acquired resistance to BRAFi and two lines with intrinsic resistance. PTK and STK activities in cell lysates were measured in the presence and absence of ex vivo BRAFi and/or MEKi. In tissue lysates, concentration-dependent <i>ex vivo</i> inhibition of STK and PTK activities with dabrafenib was stronger in responders than in non-responders. This difference was confirmed in cell lines comparing sensitive and resistant ones. Interestingly, common features of resistance were increased activity of receptor tyrosine kinases, Proto-oncogene tyrosine-protein kinase Src (Src) family kinases and protein kinase B (PKB, AKT) signalling. These latter results were confirmed by Western blots. While dabrafenib alone showed an inhibition of STK and PTK activities in both tissues and cell lines, the combination of dabrafenib and trametinib showed an antagonism on the STK activities and a synergism on PTK activities, resulting in stronger inhibitions of overall tyrosine kinase activities. Altogether; these data reveal that resistance of tumours and cell lines to MAPK inhibitors can be predicted using a multiplex kinase assay and is associated with an increase in specific tyrosine kinase activities and globally to AKT signalling in the patient’s tissue. Thus, such a predictive kinome signature would help to identify patients with innate resistance to MAPK double inhibition in order to propose other therapies.https://www.mdpi.com/2072-6694/12/2/512mapk pathwayphosphotyrosine kinaseserine-threonine kinasekinase inhibitorsintrinsic and acquired resistance |
| spellingShingle | Mohamad Krayem Philippe Aftimos Ahmad Najem Tim van den Hooven Adriënne van den Berg Liesbeth Hovestad-Bijl Rik de Wijn Riet Hilhorst Rob Ruijtenbeek Malak Sabbah Joseph Kerger Ahmad Awada Fabrice Journe Ghanem E. Ghanem Kinome Profiling to Predict Sensitivity to MAPK Inhibition in Melanoma and to Provide New Insights into Intrinsic and Acquired Mechanism of Resistance <break/>Short Title: Sensitivity Prediction to MAPK Inhibitors in Melanoma Cancers mapk pathway phosphotyrosine kinase serine-threonine kinase kinase inhibitors intrinsic and acquired resistance |
| title | Kinome Profiling to Predict Sensitivity to MAPK Inhibition in Melanoma and to Provide New Insights into Intrinsic and Acquired Mechanism of Resistance <break/>Short Title: Sensitivity Prediction to MAPK Inhibitors in Melanoma |
| title_full | Kinome Profiling to Predict Sensitivity to MAPK Inhibition in Melanoma and to Provide New Insights into Intrinsic and Acquired Mechanism of Resistance <break/>Short Title: Sensitivity Prediction to MAPK Inhibitors in Melanoma |
| title_fullStr | Kinome Profiling to Predict Sensitivity to MAPK Inhibition in Melanoma and to Provide New Insights into Intrinsic and Acquired Mechanism of Resistance <break/>Short Title: Sensitivity Prediction to MAPK Inhibitors in Melanoma |
| title_full_unstemmed | Kinome Profiling to Predict Sensitivity to MAPK Inhibition in Melanoma and to Provide New Insights into Intrinsic and Acquired Mechanism of Resistance <break/>Short Title: Sensitivity Prediction to MAPK Inhibitors in Melanoma |
| title_short | Kinome Profiling to Predict Sensitivity to MAPK Inhibition in Melanoma and to Provide New Insights into Intrinsic and Acquired Mechanism of Resistance <break/>Short Title: Sensitivity Prediction to MAPK Inhibitors in Melanoma |
| title_sort | kinome profiling to predict sensitivity to mapk inhibition in melanoma and to provide new insights into intrinsic and acquired mechanism of resistance break short title sensitivity prediction to mapk inhibitors in melanoma |
| topic | mapk pathway phosphotyrosine kinase serine-threonine kinase kinase inhibitors intrinsic and acquired resistance |
| url | https://www.mdpi.com/2072-6694/12/2/512 |
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