Lung cancer remains the leading cause of cancer deaths worldwide. International societies have pr... more Lung cancer remains the leading cause of cancer deaths worldwide. International societies have promoted the molecular analysis of MET proto-oncogene, receptor tyrosine kinase (MET) exon 14 skipping for the clinical stratification of non-small cell lung cancer (NSCLC) patients. Different technical approaches are available to detect MET exon 14 skipping in routine practice. Here, the technical performance and reproducibility of testing strategies for MET exon 14 skipping carried out in various centers were evaluated. In this retrospective study, each institution received a set (n = 10) of a customized artificial formalin-fixed paraffin-embedded (FFPE) cell line (Custom METex14 skipping FFPE block) that harbored the MET exon 14 skipping mutation (Seracare Life Sciences, Milford, MA, USA), which was previously validated by the Predictive Molecular Pathology Laboratory at the University of Naples Federico II. Each participating institution managed the reference slides according to their ...
A range of different techniques are available for predictive biomarker testing for non-small-cell... more A range of different techniques are available for predictive biomarker testing for non-small-cell lung cancer (NSCLC) clinical management. International guidelines suggest next-generation sequencing (NGS) as the preferred procedure, but other reverse transcriptase-polymerase chain reaction (RT-PCR)-based methods are rapidly evolving. In this study, we evaluated the reliability and accuracy of the IdyllaTM GeneFusion assay, a rapid and fully automated platform able to simultaneously detect ALK, ROS1, RET and NTRK1/2/3 and MET ex14 skipping mutations and compared its performance with routine reference methods. The cohort included thirty-seven NSCLCs plus two parotid gland carcinomas, previously characterized for the above alterations through either IHC, FISH, RT-PCR or NGS. In 36 of 39 cases, the Idylla GeneFusion assay and the reference methods were concordant (overall agreement: 92.3%). Tumor sections stored at room temperature for up to 60 days and 17 cases older than 2 years were ...
e20628 Background: Hyperprogressive disease (HPD) incidence ranges from 8% to 21% in patients tre... more e20628 Background: Hyperprogressive disease (HPD) incidence ranges from 8% to 21% in patients treated with anti-PD-1/PD-L1 mAbs in NSCLC and is associated with poor survival. Previously published data underlined a link between HPD across different cancers types and specific genetic alterations, such as MDM2 amplification and EGFR aberrations. We present a single-center cohort of patients with NSCLC and PD-L1 > 50% treated with 1st-line pembrolizumab. We performed NGS, IHC and FISH analysis to evaluate genetic correlations with the clinical phenotype. Methods: Clinical data from 20 patients with diagnosis of advanced NSCLC treated with 1stline immunotherapy pembrolizumab were retrospectively collected. HPD was defined by Time to Treatment Failure ≤2 months and raising in Tumor Burden ≥50% compared with basal CT-scan. MDM2 amplification was investigated by FISH on FFPE tissue sections using the MDM2/CON12 break apart FISH Probe. Positive cases were defined as those with > 10% po...
The Identification of reliable Biomarkers able to predict the outcome after nephrectomy of patien... more The Identification of reliable Biomarkers able to predict the outcome after nephrectomy of patients with clear cell renal cell carcinoma (ccRCC) is an unmet need. The gene expression analysis in tumor tissues represents a promising tool for better stratification of ccRCC subtypes and patients’ evaluation. Methods: In our study we retrospectively analyzed using Next-Generation expression analysis (NanoString), the expression of a gene panel in tumor tissue from 46 consecutive patients treated with nephrectomy for non-metastatic ccRCC at two Italian Oncological Centres. Significant differences in expression levels of selected genes was sought. Additionally, we performed a univariate and a multivariate analysis on overall survival according to Cox regression model. Results: A 17-gene expression signature of patients with a recurrence-free survival (RFS) < 1 year (unfavorable genomic signature (UGS)) and of patients with a RFS > 5 years (favorable genomic signature (FGS)) was iden...
Background: OSAS is characterized by a chronic intermittent hypoxia (CIH) and sleep fragmentation... more Background: OSAS is characterized by a chronic intermittent hypoxia (CIH) and sleep fragmentation. Polysonnography is the standard diagnostic test. New biomarkers have been proposed. The BDNF is a key mediator of neuronal and synaptic plasticity in cognitive functions. Experimental studies demonstrate changes of serum levels of BDNF following CIH Objectives: To determine serum level of BDNF in patients with severe OSAS compared with a control group and its relationship to neurocognitive function. Changes in BDNF serum levels in OSAS patients after CPAP treatment. Methods: Serum BDNF and the Montreal Cognitive Assestment (MoCA) are evaluated in 12 patients with newly diagnosed severe OSAS and in 8 healthy patients (control group). In OSAS patients serum BDNF and MoCA were reassessed after 45 days of CPAP treatment. Results: BDNF in OSAS patients was significantly higher compared to control group (p=0.01) and showed a positive correlation with ODI (p=0.01, r=0.6). Serum BDNF had a decreasing trend after CPAP treatment. Before the treatment, MoCA scores were lower in OSAS patients than controls and are positively correlated with serum BDNF levels (p=0.02, r=0.57). MoCA is normalized by CPAP therapy Conclusions: OSAS patients showed higher serum BDNF levels and lower MoCA scores compared to controls. The positive correlation between BDNF values and MoCA might reveal a peculiar BDNF-mediated neuroprotective mechanism against neuroinflammation induced by hypoxia.
Background: OSAS is characterized by a chronic intermittent hypoxia (CIH) and sleep fragmentation... more Background: OSAS is characterized by a chronic intermittent hypoxia (CIH) and sleep fragmentation. PSG is the gold standard for diagnosis. New biomarkers have been proposed. BDNF is a key mediator of cognitive functions. Changes of BDNF serum levels follow CIH. OSAS patients have mucosal upper airways inflammation. BDNF has been identified in various chronic inflammatory airways diseases. Objectives: BDNF serum levels and mucosal expression in severe OSAS patients compared to controls. Relationship of serum BDNF with neurocognitive function. Changes in serum BDNF after CPAP. Methods: Serum BDNF, upper airways mucosal biopsy and Montreal Cognitive Assestment (MoCA) are performed in 12 naive severe OSAS patients and in 8 controls. In OSAS patients serum BDNF and MoCA were reassessed after 45 days of CPAP treatment. Results: BDNF in OSAS was significantly higher than controls (p=0.01) and showed a positive correlation with ODI (p=0.01, r=0.6). Serum BDNF had a decreasing trend after CPAP treatment. Before the treatment, MoCA scores were lower in OSAS than controls and are positively correlated with serum BDNF levels (p=0.02, r=0.57). MoCA is normalized by CPAP therapy. Immunohistochemistry of mucosal biopsy in OSAS patients showed higher inflammatory infiltrate CD3+ CD79a+ (BDNF positive) than controls. Conclusions: OSAS patients showed higher serum BDNF levels and lower MoCA scores than controls. The positive correlation between BDNF values and MoCA might reveal a peculiar BDNF-mediated neuroprotective mechanism against neuroinflammation induced by hypoxia. OSAS have mucosal upper airway lymphocytic inflammation BDNF positive.
2659 Background: Gene-expression-profiling defined at least two main groups within Diffuse-large-... more 2659 Background: Gene-expression-profiling defined at least two main groups within Diffuse-large-B-cell Lymphoma (DLBCL) patients who have substantially different outcomes: Activated-B-cell (ABC-type) and Germinal-Center-B-cell (GCB-type). The translation of gene-expression-profiling arrays into robust algorithm useful for clinical purposes is still in progress. The detection of IgM monoclonal component (IgM MC) in DLBCL has been previously described in a few reports, mainly because it was associated with autoimmune hemolytic anaemia. To our knowledge this is the first report describing the incidence and prognosis of a series of DLBCL with IgM MC. Aims: In this report we compared clinical and biological features of DLBCL patients with and without secretory IgM MC . Patients…
Hypomethylation has been reported to be responsible for the activation of several oncogenes. The ... more Hypomethylation has been reported to be responsible for the activation of several oncogenes. The possibility that hypomethylation is involved in the regulation of MET transcription was investigated through the analysis of the methylation status of one CpG island containing 43 CpGs in six cases of papillary carcinoma, in the corresponding normal thyroid tissue, and in two cases of hyperplastic goitre. Evidence of methylation was not found in any of the analysed CpG.
Met protein, the high affinity receptor for hepatocyte growth factor (HGF), was highly expressed ... more Met protein, the high affinity receptor for hepatocyte growth factor (HGF), was highly expressed by the tumour cells of 64 well-differentiated papillary carcinomas of the thyroid. The p145 mature form and the p170 precursor form of the protein were both isolated from the tumours. Enhanced expression of Met protein was associated with a 9.5 +/- 5-fold increase in MET RNA transcript levels, suggesting increased transcription of the gene. In the same tumours, the levels of RNA transcripts for hypoxia inducible factor-1 (HIF-1), a potent stimulator of met gene transcription, were 4.5 +/- 3-fold higher than those present in the surrounding normal thyroid tissues. HIF-1 is generally induced by hypoxia. Histological features suggestive of a hypoxia were observed in 37 of 50 tumours and included coagulative necrosis, psammoma bodies, cystic changes, intratumoural haemorrhage, and hyalinization of the fibrous stroma. Immunostaining for Met protein was particularly intense in some cells located at the tumour periphery which were characterized by an invasive phenotype. Microdissection of tumour cell nests from the invading front revealed that the levels of RNA transcripts for MET/HIF were higher than in the centre of the tumour in four of nine cases. Taken together, the findings of this study suggest that HIF-1, perhaps driven by hypoxia, may be one of the factors leading to the increased transcription of met gene in papillary carcinoma and that this event is often more pronounced at the tumour periphery.
The present study has investigated the functional role of the Met receptor in primary cultures of... more The present study has investigated the functional role of the Met receptor in primary cultures of 20 papillary carcinomas and of normal thyroid cells obtained from the same patients. Normal and tumour cells grew as adherent cells, formed a confluent monolayer after 10-20 days, had epithelial morphology, and were immunoreactive for cytokeratin, vimentin, and thyroglobulin. The potential effect of hepatocyte growth factor (HGF) on cell invasiveness was investigated in Boyden chambers, using a nucleopore filter coated with Matrigel as the barrier and HGF as the chemoattractant. Tumour cells of five out of seven cases of papillary carcinoma were more responsive to HGF than the corresponding normal cells in terms of the number of migrated cells per mm(2). Involvement of the Met receptor in the HGF-induced migratory response was suggested by the observation that the agonistic anti-Met monoclonal antibody (MAb) DO-24 was equally effective. HGF did not affect the proliferative activity of thyroid cells. Under the same experimental conditions, 10 per cent fetal bovine serum (FBS) induced a two-fold increase in [(3)H]thymidine incorporation into normal cells and tumour cells. These findings are consistent with the possibility that HGF plays a crucial role in determining the invasiveness of tumour cells in papillary carcinoma of the thyroid.
Lung cancer remains the leading cause of cancer deaths worldwide. International societies have pr... more Lung cancer remains the leading cause of cancer deaths worldwide. International societies have promoted the molecular analysis of MET proto-oncogene, receptor tyrosine kinase (MET) exon 14 skipping for the clinical stratification of non-small cell lung cancer (NSCLC) patients. Different technical approaches are available to detect MET exon 14 skipping in routine practice. Here, the technical performance and reproducibility of testing strategies for MET exon 14 skipping carried out in various centers were evaluated. In this retrospective study, each institution received a set (n = 10) of a customized artificial formalin-fixed paraffin-embedded (FFPE) cell line (Custom METex14 skipping FFPE block) that harbored the MET exon 14 skipping mutation (Seracare Life Sciences, Milford, MA, USA), which was previously validated by the Predictive Molecular Pathology Laboratory at the University of Naples Federico II. Each participating institution managed the reference slides according to their ...
A range of different techniques are available for predictive biomarker testing for non-small-cell... more A range of different techniques are available for predictive biomarker testing for non-small-cell lung cancer (NSCLC) clinical management. International guidelines suggest next-generation sequencing (NGS) as the preferred procedure, but other reverse transcriptase-polymerase chain reaction (RT-PCR)-based methods are rapidly evolving. In this study, we evaluated the reliability and accuracy of the IdyllaTM GeneFusion assay, a rapid and fully automated platform able to simultaneously detect ALK, ROS1, RET and NTRK1/2/3 and MET ex14 skipping mutations and compared its performance with routine reference methods. The cohort included thirty-seven NSCLCs plus two parotid gland carcinomas, previously characterized for the above alterations through either IHC, FISH, RT-PCR or NGS. In 36 of 39 cases, the Idylla GeneFusion assay and the reference methods were concordant (overall agreement: 92.3%). Tumor sections stored at room temperature for up to 60 days and 17 cases older than 2 years were ...
e20628 Background: Hyperprogressive disease (HPD) incidence ranges from 8% to 21% in patients tre... more e20628 Background: Hyperprogressive disease (HPD) incidence ranges from 8% to 21% in patients treated with anti-PD-1/PD-L1 mAbs in NSCLC and is associated with poor survival. Previously published data underlined a link between HPD across different cancers types and specific genetic alterations, such as MDM2 amplification and EGFR aberrations. We present a single-center cohort of patients with NSCLC and PD-L1 > 50% treated with 1st-line pembrolizumab. We performed NGS, IHC and FISH analysis to evaluate genetic correlations with the clinical phenotype. Methods: Clinical data from 20 patients with diagnosis of advanced NSCLC treated with 1stline immunotherapy pembrolizumab were retrospectively collected. HPD was defined by Time to Treatment Failure ≤2 months and raising in Tumor Burden ≥50% compared with basal CT-scan. MDM2 amplification was investigated by FISH on FFPE tissue sections using the MDM2/CON12 break apart FISH Probe. Positive cases were defined as those with > 10% po...
The Identification of reliable Biomarkers able to predict the outcome after nephrectomy of patien... more The Identification of reliable Biomarkers able to predict the outcome after nephrectomy of patients with clear cell renal cell carcinoma (ccRCC) is an unmet need. The gene expression analysis in tumor tissues represents a promising tool for better stratification of ccRCC subtypes and patients’ evaluation. Methods: In our study we retrospectively analyzed using Next-Generation expression analysis (NanoString), the expression of a gene panel in tumor tissue from 46 consecutive patients treated with nephrectomy for non-metastatic ccRCC at two Italian Oncological Centres. Significant differences in expression levels of selected genes was sought. Additionally, we performed a univariate and a multivariate analysis on overall survival according to Cox regression model. Results: A 17-gene expression signature of patients with a recurrence-free survival (RFS) < 1 year (unfavorable genomic signature (UGS)) and of patients with a RFS > 5 years (favorable genomic signature (FGS)) was iden...
Background: OSAS is characterized by a chronic intermittent hypoxia (CIH) and sleep fragmentation... more Background: OSAS is characterized by a chronic intermittent hypoxia (CIH) and sleep fragmentation. Polysonnography is the standard diagnostic test. New biomarkers have been proposed. The BDNF is a key mediator of neuronal and synaptic plasticity in cognitive functions. Experimental studies demonstrate changes of serum levels of BDNF following CIH Objectives: To determine serum level of BDNF in patients with severe OSAS compared with a control group and its relationship to neurocognitive function. Changes in BDNF serum levels in OSAS patients after CPAP treatment. Methods: Serum BDNF and the Montreal Cognitive Assestment (MoCA) are evaluated in 12 patients with newly diagnosed severe OSAS and in 8 healthy patients (control group). In OSAS patients serum BDNF and MoCA were reassessed after 45 days of CPAP treatment. Results: BDNF in OSAS patients was significantly higher compared to control group (p=0.01) and showed a positive correlation with ODI (p=0.01, r=0.6). Serum BDNF had a decreasing trend after CPAP treatment. Before the treatment, MoCA scores were lower in OSAS patients than controls and are positively correlated with serum BDNF levels (p=0.02, r=0.57). MoCA is normalized by CPAP therapy Conclusions: OSAS patients showed higher serum BDNF levels and lower MoCA scores compared to controls. The positive correlation between BDNF values and MoCA might reveal a peculiar BDNF-mediated neuroprotective mechanism against neuroinflammation induced by hypoxia.
Background: OSAS is characterized by a chronic intermittent hypoxia (CIH) and sleep fragmentation... more Background: OSAS is characterized by a chronic intermittent hypoxia (CIH) and sleep fragmentation. PSG is the gold standard for diagnosis. New biomarkers have been proposed. BDNF is a key mediator of cognitive functions. Changes of BDNF serum levels follow CIH. OSAS patients have mucosal upper airways inflammation. BDNF has been identified in various chronic inflammatory airways diseases. Objectives: BDNF serum levels and mucosal expression in severe OSAS patients compared to controls. Relationship of serum BDNF with neurocognitive function. Changes in serum BDNF after CPAP. Methods: Serum BDNF, upper airways mucosal biopsy and Montreal Cognitive Assestment (MoCA) are performed in 12 naive severe OSAS patients and in 8 controls. In OSAS patients serum BDNF and MoCA were reassessed after 45 days of CPAP treatment. Results: BDNF in OSAS was significantly higher than controls (p=0.01) and showed a positive correlation with ODI (p=0.01, r=0.6). Serum BDNF had a decreasing trend after CPAP treatment. Before the treatment, MoCA scores were lower in OSAS than controls and are positively correlated with serum BDNF levels (p=0.02, r=0.57). MoCA is normalized by CPAP therapy. Immunohistochemistry of mucosal biopsy in OSAS patients showed higher inflammatory infiltrate CD3+ CD79a+ (BDNF positive) than controls. Conclusions: OSAS patients showed higher serum BDNF levels and lower MoCA scores than controls. The positive correlation between BDNF values and MoCA might reveal a peculiar BDNF-mediated neuroprotective mechanism against neuroinflammation induced by hypoxia. OSAS have mucosal upper airway lymphocytic inflammation BDNF positive.
2659 Background: Gene-expression-profiling defined at least two main groups within Diffuse-large-... more 2659 Background: Gene-expression-profiling defined at least two main groups within Diffuse-large-B-cell Lymphoma (DLBCL) patients who have substantially different outcomes: Activated-B-cell (ABC-type) and Germinal-Center-B-cell (GCB-type). The translation of gene-expression-profiling arrays into robust algorithm useful for clinical purposes is still in progress. The detection of IgM monoclonal component (IgM MC) in DLBCL has been previously described in a few reports, mainly because it was associated with autoimmune hemolytic anaemia. To our knowledge this is the first report describing the incidence and prognosis of a series of DLBCL with IgM MC. Aims: In this report we compared clinical and biological features of DLBCL patients with and without secretory IgM MC . Patients…
Hypomethylation has been reported to be responsible for the activation of several oncogenes. The ... more Hypomethylation has been reported to be responsible for the activation of several oncogenes. The possibility that hypomethylation is involved in the regulation of MET transcription was investigated through the analysis of the methylation status of one CpG island containing 43 CpGs in six cases of papillary carcinoma, in the corresponding normal thyroid tissue, and in two cases of hyperplastic goitre. Evidence of methylation was not found in any of the analysed CpG.
Met protein, the high affinity receptor for hepatocyte growth factor (HGF), was highly expressed ... more Met protein, the high affinity receptor for hepatocyte growth factor (HGF), was highly expressed by the tumour cells of 64 well-differentiated papillary carcinomas of the thyroid. The p145 mature form and the p170 precursor form of the protein were both isolated from the tumours. Enhanced expression of Met protein was associated with a 9.5 +/- 5-fold increase in MET RNA transcript levels, suggesting increased transcription of the gene. In the same tumours, the levels of RNA transcripts for hypoxia inducible factor-1 (HIF-1), a potent stimulator of met gene transcription, were 4.5 +/- 3-fold higher than those present in the surrounding normal thyroid tissues. HIF-1 is generally induced by hypoxia. Histological features suggestive of a hypoxia were observed in 37 of 50 tumours and included coagulative necrosis, psammoma bodies, cystic changes, intratumoural haemorrhage, and hyalinization of the fibrous stroma. Immunostaining for Met protein was particularly intense in some cells located at the tumour periphery which were characterized by an invasive phenotype. Microdissection of tumour cell nests from the invading front revealed that the levels of RNA transcripts for MET/HIF were higher than in the centre of the tumour in four of nine cases. Taken together, the findings of this study suggest that HIF-1, perhaps driven by hypoxia, may be one of the factors leading to the increased transcription of met gene in papillary carcinoma and that this event is often more pronounced at the tumour periphery.
The present study has investigated the functional role of the Met receptor in primary cultures of... more The present study has investigated the functional role of the Met receptor in primary cultures of 20 papillary carcinomas and of normal thyroid cells obtained from the same patients. Normal and tumour cells grew as adherent cells, formed a confluent monolayer after 10-20 days, had epithelial morphology, and were immunoreactive for cytokeratin, vimentin, and thyroglobulin. The potential effect of hepatocyte growth factor (HGF) on cell invasiveness was investigated in Boyden chambers, using a nucleopore filter coated with Matrigel as the barrier and HGF as the chemoattractant. Tumour cells of five out of seven cases of papillary carcinoma were more responsive to HGF than the corresponding normal cells in terms of the number of migrated cells per mm(2). Involvement of the Met receptor in the HGF-induced migratory response was suggested by the observation that the agonistic anti-Met monoclonal antibody (MAb) DO-24 was equally effective. HGF did not affect the proliferative activity of thyroid cells. Under the same experimental conditions, 10 per cent fetal bovine serum (FBS) induced a two-fold increase in [(3)H]thymidine incorporation into normal cells and tumour cells. These findings are consistent with the possibility that HGF plays a crucial role in determining the invasiveness of tumour cells in papillary carcinoma of the thyroid.
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Papers by Stefania Scarpino