The c-Met receptor is a therapeutically actionable target in non-small-cell lung cancer (NSCLC), ... more The c-Met receptor is a therapeutically actionable target in non-small-cell lung cancer (NSCLC), with one approved drug and several agents in development. Most suitable biomarkers for patient selection include c-Met amplification and exon-14 skipping. Our retrospective study focused on the frequency of different c-Met aberrations (overexpression, amplification and mutations) in 153 primary, therapy-naïve resection samples and their paired metastases, from Biobank@UZA. Furthermore, we determined the correlation of c-Met expression with clinicopathological factors, Epidermal Growth Factor Receptor (EGFR)-status and TP53 mutations. Our results showed that c-Met expression levels in primary tumors were comparable to their respective metastases. Five different mutations were detected by deep sequencing: three (E168D, S203T, N375S) previously described and two never reported (I333T, G783E). I333T, a new mutation in the Sema(phorin) domain of c-Met, might influence the binding of antibodie...
Several oncogenic drivers have been identified in non-small cell lung cancer. Targeted therapies ... more Several oncogenic drivers have been identified in non-small cell lung cancer. Targeted therapies for these aberrations have already been successfully developed and implemented in clinical practice. Owing to improved sensitivity in genetic testing, more and more tumors with multiple driver mutations are identified, resulting in dilemmas for treating physicians whether and which targeted therapy to use. In this case series, we provide an overview of patients with intrinsic double mutations in oncogenic drivers and their reported response to targeted therapies, with a focus on epidermal growth factor receptor, anaplastic lymphoma kinase, cMET, and Kirsten rat sarcoma viral oncogene. We also include an unpublished case report on a patient with an epidermal growth factor receptor L858R and cMET exon 14 skipping.
Erlotinib is commonly used as a second line treatment in non-small cell lung cancer patients with... more Erlotinib is commonly used as a second line treatment in non-small cell lung cancer patients with sensitizing EGFR mutations. In EGFR-wild type patients, however the results are limited. Therefore we evaluated whether the combination of the Protein kinase C-β inhibitor enzastaurin with erlotinib could enhance the effect in the A549 and H1650 cell lines. Cytotoxicity of erlotinib, enzastaurin and their 72-h simultaneous combination was assessed with the MTT assay. The pharmacologic interaction was studied using the method of Chou and Talalay, cell cycle perturbations were assessed by flow cytometry and modulation of ERK1/2 and AKT phosphorylation was determined with ELISA. For protein phosphorylation of GSK3β we performed Western Blot analysis and a Pamgene phosphorylation array, while RT-PCR was used to investigate VEGF and VEGFR-2 expression before and after drug treatments. A synergistic interaction was found in both cell lines with mean CI of 0.58 and 0.63 in A549 and H1650 cells...
Advances in experimental medicine and biology, 2017
Lung cancer is the leader malignancy worldwide accounting 1.5 millions of deaths every year. In t... more Lung cancer is the leader malignancy worldwide accounting 1.5 millions of deaths every year. In the United States the 5 year-overall survival is less than 20% for all the newly diagnosed patients. Cisplatin-based cytotoxic chemotherapy for unresectable or metastatic NSCLC patients in the first line of treatment, and docetaxel in the second line, have achieved positive results but with limited benefit in overall survival. Targeted therapies for EGFR and ALK mutant patients have showed better results when compared with chemotherapy, nevertheless most of patients will fail and need to be treated with chemotherapy if they still have a good performance status.Immunotherapy recently has become the most revolutionary treatment in solid tumors patients. First results in unresectable and metastatic melanoma patients treated with an anti CTLA-4 monoclonal antibody showed an unexpected 3-year overall survival of at least 25%.Lung cancer cells have multiple immunosuppressive mechanisms that all...
Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, Sep 17, 2016
The abnormal stimulation of the multiple signal transduction pathways downstream of the receptor ... more The abnormal stimulation of the multiple signal transduction pathways downstream of the receptor tyrosine kinase mesenchymal-epithelial transition factor (cMET) promotes cellular transformation, tumor motility and invasion. Therefore, cMET has been the focus of prognostic and therapeutic studies in different tumor types, including non-small cell lung cancer (NSCLC). In particular, several cMET-inhibitors have been developed as innovative therapeutic candidates, and are currently under investigation in clinical trials. However, one of the challenges in establishing effective targeted treatments against cMET, remains the accurate identification of biomarkers for the selection of responsive subsets of patients. Recently, splice site mutations have been discovered in cMET that lead to the skipping of exon 14, impairing the break-down of the receptor. NSCLC-patients carrying this splice variant typically overexpress the cMET receptor and show a response to small molecule inhibitors of cM...
Introduction There are numerous causes triggering CRC. 25-80% of CRC shown a deregulation in Epid... more Introduction There are numerous causes triggering CRC. 25-80% of CRC shown a deregulation in Epidermal Growth Factor Receptor (EGFR) pathway. Two signaling pathways downstream of the EGFR are dysregulated in CRC the mitogen-activated protein kinase (MAPK) and the phosphoinositide-3-kinase (PI3K) pathway. Activating mutations in KRAS and BRAF (MAPK pathway) and PIK3CA affect prognosis and/or response to anti-EGFR MoAb. PTEN is a downstream effector of EGFR pathway and is involved in PI3K pathway. Loss of PTEN protein expression can occur through epigenetic silencing and mutation or allelic loss. Immunohistochemistry (IHC) is the most effective way to assay for loss of PTEN expression. IHC lack of reproducibility and this has lead to discordant results regarding the concordant in paired CRC metastases and primary tumors.The aim of the work was to asses the feasibility of PTEN gene expression analysis from FFPE tissue in CRC samples. Materials and methods We selected a series of 33 pat...
The last few decades have witnessed a silent revolution in the war against NSCLC, thanks to the d... more The last few decades have witnessed a silent revolution in the war against NSCLC, thanks to the discovery of "oncogenic drivers" and the subsequent development of targeted therapies. The discovery of the EML4-ALK fusion gene in a subgroup of patients with NSCLC and the subsequent clinical development of crizotinib has been an amazing success story in lung cancer translational-research, and its accelerated approval [only 4 years from the discovery of ALK rearrangement in NSCLC to the approval by the Food and Drug Administration (FDA)] marked the beginning of the new decade of targeted therapy. However, common to all targeted therapies, despite an initial benefit, patients inevitably experience tumor progression, due to the development of resistance. Several molecular mechanisms are responsible for acquired resistance, such as secondary mutations of ALK kinase domain or amplification of ALK fusion gene, or the activation of other oncogenic drivers, which may cause resistance...
Journal of the American Board of Family Medicine : JABFM
In the past decade the advent of target therapy has led to a silent revolution in the treatment o... more In the past decade the advent of target therapy has led to a silent revolution in the treatment of lung cancer. Thanks to the specificity of their target, new tailored drugs are able to achieve a larger benefit and lower toxicity and provide better quality of life than cytotoxic drugs in a limited number of patients, selected by molecular profile. Nowadays, the epidermal growth factor receptor tyrosine kinase inhibitors erlotinib and gefitinib, and the anaplastic lymphoma kinase inhibitor crizotinib, are targeted agents approved for treatment of non-small-cell lung cancer. Family physicians play an important role in the treatment, detection, and management of common toxicities and in providing emotional support. Therefore this review integrates molecular profile assessment with evidence of the efficacy and toxicity of tyrosine kinase inhibitors to provide an updated overview of the treatment of non-small-cell lung cancer, which radically changed after the advent of targeted therapie...
ABSTRACT Introduction: Despite recent advances, lung cancer shows a poor prognosis and a high inc... more ABSTRACT Introduction: Despite recent advances, lung cancer shows a poor prognosis and a high incidence of recurrence. Today, targeted therapies against the epidermal growth factor receptor (EGFR) are frequently used in the clinic. cMET amplification is a known resistance mechanism against these EGFR-targeted therapies. The current study determines the influence of cMET amplification, expression and autocrine stimulation, as well as EGFR mutational status and expression on the cytotoxicity of small molecule inhibitors.Conclusion: These results show that cMET amplification confers the greatest sensitivity to cMET-TKIs and to Hsp90 inhibition, where a sensitizing mutation in EGFR leads to great sensitivity to erlotinib. The EGFR-dependent cell lines HCC827 and H1975 also show moderate sensitivity to Hsp90 inhibition. HGF-secretion does not lead to great sensitivity to cMET-inhibition or Hsp90-inhibition. Based on these results, combination therapy can be tested with cMET- and EGFR-TKI.
The c-Met receptor is a therapeutically actionable target in non-small-cell lung cancer (NSCLC), ... more The c-Met receptor is a therapeutically actionable target in non-small-cell lung cancer (NSCLC), with one approved drug and several agents in development. Most suitable biomarkers for patient selection include c-Met amplification and exon-14 skipping. Our retrospective study focused on the frequency of different c-Met aberrations (overexpression, amplification and mutations) in 153 primary, therapy-naïve resection samples and their paired metastases, from Biobank@UZA. Furthermore, we determined the correlation of c-Met expression with clinicopathological factors, Epidermal Growth Factor Receptor (EGFR)-status and TP53 mutations. Our results showed that c-Met expression levels in primary tumors were comparable to their respective metastases. Five different mutations were detected by deep sequencing: three (E168D, S203T, N375S) previously described and two never reported (I333T, G783E). I333T, a new mutation in the Sema(phorin) domain of c-Met, might influence the binding of antibodie...
Several oncogenic drivers have been identified in non-small cell lung cancer. Targeted therapies ... more Several oncogenic drivers have been identified in non-small cell lung cancer. Targeted therapies for these aberrations have already been successfully developed and implemented in clinical practice. Owing to improved sensitivity in genetic testing, more and more tumors with multiple driver mutations are identified, resulting in dilemmas for treating physicians whether and which targeted therapy to use. In this case series, we provide an overview of patients with intrinsic double mutations in oncogenic drivers and their reported response to targeted therapies, with a focus on epidermal growth factor receptor, anaplastic lymphoma kinase, cMET, and Kirsten rat sarcoma viral oncogene. We also include an unpublished case report on a patient with an epidermal growth factor receptor L858R and cMET exon 14 skipping.
Erlotinib is commonly used as a second line treatment in non-small cell lung cancer patients with... more Erlotinib is commonly used as a second line treatment in non-small cell lung cancer patients with sensitizing EGFR mutations. In EGFR-wild type patients, however the results are limited. Therefore we evaluated whether the combination of the Protein kinase C-β inhibitor enzastaurin with erlotinib could enhance the effect in the A549 and H1650 cell lines. Cytotoxicity of erlotinib, enzastaurin and their 72-h simultaneous combination was assessed with the MTT assay. The pharmacologic interaction was studied using the method of Chou and Talalay, cell cycle perturbations were assessed by flow cytometry and modulation of ERK1/2 and AKT phosphorylation was determined with ELISA. For protein phosphorylation of GSK3β we performed Western Blot analysis and a Pamgene phosphorylation array, while RT-PCR was used to investigate VEGF and VEGFR-2 expression before and after drug treatments. A synergistic interaction was found in both cell lines with mean CI of 0.58 and 0.63 in A549 and H1650 cells...
Advances in experimental medicine and biology, 2017
Lung cancer is the leader malignancy worldwide accounting 1.5 millions of deaths every year. In t... more Lung cancer is the leader malignancy worldwide accounting 1.5 millions of deaths every year. In the United States the 5 year-overall survival is less than 20% for all the newly diagnosed patients. Cisplatin-based cytotoxic chemotherapy for unresectable or metastatic NSCLC patients in the first line of treatment, and docetaxel in the second line, have achieved positive results but with limited benefit in overall survival. Targeted therapies for EGFR and ALK mutant patients have showed better results when compared with chemotherapy, nevertheless most of patients will fail and need to be treated with chemotherapy if they still have a good performance status.Immunotherapy recently has become the most revolutionary treatment in solid tumors patients. First results in unresectable and metastatic melanoma patients treated with an anti CTLA-4 monoclonal antibody showed an unexpected 3-year overall survival of at least 25%.Lung cancer cells have multiple immunosuppressive mechanisms that all...
Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer, Sep 17, 2016
The abnormal stimulation of the multiple signal transduction pathways downstream of the receptor ... more The abnormal stimulation of the multiple signal transduction pathways downstream of the receptor tyrosine kinase mesenchymal-epithelial transition factor (cMET) promotes cellular transformation, tumor motility and invasion. Therefore, cMET has been the focus of prognostic and therapeutic studies in different tumor types, including non-small cell lung cancer (NSCLC). In particular, several cMET-inhibitors have been developed as innovative therapeutic candidates, and are currently under investigation in clinical trials. However, one of the challenges in establishing effective targeted treatments against cMET, remains the accurate identification of biomarkers for the selection of responsive subsets of patients. Recently, splice site mutations have been discovered in cMET that lead to the skipping of exon 14, impairing the break-down of the receptor. NSCLC-patients carrying this splice variant typically overexpress the cMET receptor and show a response to small molecule inhibitors of cM...
Introduction There are numerous causes triggering CRC. 25-80% of CRC shown a deregulation in Epid... more Introduction There are numerous causes triggering CRC. 25-80% of CRC shown a deregulation in Epidermal Growth Factor Receptor (EGFR) pathway. Two signaling pathways downstream of the EGFR are dysregulated in CRC the mitogen-activated protein kinase (MAPK) and the phosphoinositide-3-kinase (PI3K) pathway. Activating mutations in KRAS and BRAF (MAPK pathway) and PIK3CA affect prognosis and/or response to anti-EGFR MoAb. PTEN is a downstream effector of EGFR pathway and is involved in PI3K pathway. Loss of PTEN protein expression can occur through epigenetic silencing and mutation or allelic loss. Immunohistochemistry (IHC) is the most effective way to assay for loss of PTEN expression. IHC lack of reproducibility and this has lead to discordant results regarding the concordant in paired CRC metastases and primary tumors.The aim of the work was to asses the feasibility of PTEN gene expression analysis from FFPE tissue in CRC samples. Materials and methods We selected a series of 33 pat...
The last few decades have witnessed a silent revolution in the war against NSCLC, thanks to the d... more The last few decades have witnessed a silent revolution in the war against NSCLC, thanks to the discovery of "oncogenic drivers" and the subsequent development of targeted therapies. The discovery of the EML4-ALK fusion gene in a subgroup of patients with NSCLC and the subsequent clinical development of crizotinib has been an amazing success story in lung cancer translational-research, and its accelerated approval [only 4 years from the discovery of ALK rearrangement in NSCLC to the approval by the Food and Drug Administration (FDA)] marked the beginning of the new decade of targeted therapy. However, common to all targeted therapies, despite an initial benefit, patients inevitably experience tumor progression, due to the development of resistance. Several molecular mechanisms are responsible for acquired resistance, such as secondary mutations of ALK kinase domain or amplification of ALK fusion gene, or the activation of other oncogenic drivers, which may cause resistance...
Journal of the American Board of Family Medicine : JABFM
In the past decade the advent of target therapy has led to a silent revolution in the treatment o... more In the past decade the advent of target therapy has led to a silent revolution in the treatment of lung cancer. Thanks to the specificity of their target, new tailored drugs are able to achieve a larger benefit and lower toxicity and provide better quality of life than cytotoxic drugs in a limited number of patients, selected by molecular profile. Nowadays, the epidermal growth factor receptor tyrosine kinase inhibitors erlotinib and gefitinib, and the anaplastic lymphoma kinase inhibitor crizotinib, are targeted agents approved for treatment of non-small-cell lung cancer. Family physicians play an important role in the treatment, detection, and management of common toxicities and in providing emotional support. Therefore this review integrates molecular profile assessment with evidence of the efficacy and toxicity of tyrosine kinase inhibitors to provide an updated overview of the treatment of non-small-cell lung cancer, which radically changed after the advent of targeted therapie...
ABSTRACT Introduction: Despite recent advances, lung cancer shows a poor prognosis and a high inc... more ABSTRACT Introduction: Despite recent advances, lung cancer shows a poor prognosis and a high incidence of recurrence. Today, targeted therapies against the epidermal growth factor receptor (EGFR) are frequently used in the clinic. cMET amplification is a known resistance mechanism against these EGFR-targeted therapies. The current study determines the influence of cMET amplification, expression and autocrine stimulation, as well as EGFR mutational status and expression on the cytotoxicity of small molecule inhibitors.Conclusion: These results show that cMET amplification confers the greatest sensitivity to cMET-TKIs and to Hsp90 inhibition, where a sensitizing mutation in EGFR leads to great sensitivity to erlotinib. The EGFR-dependent cell lines HCC827 and H1975 also show moderate sensitivity to Hsp90 inhibition. HGF-secretion does not lead to great sensitivity to cMET-inhibition or Hsp90-inhibition. Based on these results, combination therapy can be tested with cMET- and EGFR-TKI.
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