Cell Biology and Biotherapy Unit

Napoli, Italy

Cell Biology and Biotherapy Unit

Napoli, Italy
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Normanno N.,Cell Biology and Biotherapy Unit | Normanno N.,Crom Centro Ricerche Oncologiche Of Mercogliano | Pinto C.,S Orsola Malpighi Hospital | Taddei G.,University of Florence | And 4 more authors.
Journal of Thoracic Oncology | Year: 2013

Introduction: The Italian Association of Medical Oncology (AIOM) and the Italian Society of Pathology and Cytology organized an external quality assessment (EQA) scheme for EGFR mutation testing in non-small-cell lung cancer. Methods: Ten specimens, including three small biopsies with known epidermal growth factor receptor (EGFR) mutation status, were validated in three referral laboratories and provided to 47 participating centers. The participants were requested to perform mutational analysis, using their usual method, and to submit results within a 4-week time frame. According to a predefined scoring system, two points were assigned to correct genotype and zero points to false-negative or false-positive results. The threshold to pass the EQA was set at higher than 18 of 20 points. Two rounds were preplanned. Results: All participating centers submitted the results within the time frame. Polymerase chain reaction (PCR)/sequencing was the main methodology used (n = 37 laboratories), although a few centers did use pyrosequencing (n = 8) or real-time PCR (n = 2). A significant number of analytical errors were observed (n = 20), with a high frequency of false-positive results (n = 16). The lower scores were obtained for the small biopsies. Fourteen of 47 centers (30%) that did not pass the first round, having a score less than or equal to 18 points, used PCR/sequencing, whereas 10 of 10 laboratories, using pyrosequencing or real-time PCR, passed the first round. Eight laboratories passed the second round. Overall, 41of 47 centers (87%) passed the EQA. Conclusion: The results of the EQA for EGFR testing in non-smallcell lung cancer suggest that good quality EGFR mutational analysis is performed in Italian laboratories, although differences between testing methods were observed, especially for small biopsies. Copyright ©2013 by the International Association for the Study of Lung Cancer.


Strizzi L.,Northwestern University | Margaryan N.V.,Northwestern University | Gilgur A.,Northwestern University | Hardy K.M.,Northwestern University | And 3 more authors.
Cell Cycle | Year: 2013

Cripto-1 (CR-1) protein function differs according to cellular or extracellular expression. In this study, we explore the significance of cell surface CR-1 expression in human melanoma cells. Cell surface CR-1-expressing human melanoma cells (CR1-CS+) were selected by fluorescence-activated cell sorting (FACS) and grown in vitro and in vivo in nude mice to study their growth characteristics. The CR1-CS+ melanoma cells were found to express increased levels of Oct4, MDR-1 and activated c-Src compared with cells lacking this subpopulation (CR1-CS-) or unsorted cells, used as control. CR1-CS+ show reduced proliferation rates and diminished spherical colony formation compared with control cells when cultured in vitro. Orthotopic injections of CR1-CS+ in nude mice formed slow growing tumors with histologic variability across different areas of the CR1-CS+ xenografts. CR-1-expressing cells from first generation CR1-CS+ tumors showed significantly increased tumor-forming rate and aggressiveness following subsequent transplants in nude mice. These data demonstrate that within a heterogeneous melanoma cell population there resides a slow proliferating, cell surface CR-1-expressing subpopulation capable of giving rise to a fast growing, aggressive progeny that may contribute to disease recurrence and progression. © 2013 Landes Bioscience.


Vecchione L.,University Hospital Gasthuisberg | Vecchione L.,Catholic University of Leuven | Vecchione L.,The Second University of Naples | Jacobs B.,University Hospital Gasthuisberg | And 5 more authors.
Experimental Cell Research | Year: 2011

Anti-Epidermal Growth Factor Receptor (EGFR) therapies have been recently developed for the treatment of multiple cancer types. At the time when they were introduced in clinical practice, there was little knowledge of the molecular bases of tumor sensitivity and resistance to these novel targeted compounds. By using the framework of anti-EGFR inhibitors as treatment for colorectal cancer patients, we will review the knowledge we have reached until now in improving the development of a personalized cancer therapy and we will try to indicate the future challenges this field will face in the future. © 2011 Elsevier Inc.


van Krieken J.H.,Radboud University Nijmegen | Siebers A.G.,Radboud University Nijmegen | Normanno N.,Cell Biology and Biotherapy Unit
Annals of Oncology | Year: 2013

Molecular testing of tumor samples to guide treatment decisions is of increasing importance. Several drugs have been approved for treatment of molecularly defined subgroups of patients, and the number of agents requiring companion diagnostics for their prescription is expected to rapidly increase. The results of such testing directly influence the management of individual patients, with both false-negative and false-positive results being harmful for patients. In this respect, external quality assurance (EQA) programs are essential to guarantee optimal quality of testing. There are several EQA schemes available in Europe, but they vary in scope, size and execution. During a conference held in early 2012, medical oncologists, pathologists, geneticists, molecular biologists, EQA providers and representatives from pharmaceutical industries developed a guideline to harmonize the standards applied by EQA schemes in molecular pathology. The guideline comprises recommendations on the organization of an EQA scheme, defining the criteria for reference laboratories, requirements for EQA test samples and the number of samples that are needed for an EQA scheme. Furthermore, a scoring system is proposed and consequences of poor performance are formulated. Lastly, the contents of an EQA report, communication of the EQA results, EQA databases and participant manual are given. © The Author 2013. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved.


Ciardiello F.,The Second University of Naples | Normanno N.,Cell Biology and Biotherapy Unit
Cancer Discovery | Year: 2011

Primary and acquired resistance to anti-epidermal growth factor receptor (EGFR) drugs are clinically relevant problems in patients with metastatic colorectal carcinoma. A complex network of molecular alterations is involved in this phenomenon. Bertotti et al. report the development of serially transplantable groups of tumor xenografts in immune-deficient mice from patient-derived, genetically characterized metastatic colorectal carcinoma samples. These experimental models ("xenopatients") might represent a novel approach to discover and characterize the mechanisms of resistance to anti-EGFR therapy and other molecularly targeted therapies in metastatic colorectal carcinoma. In this respect, Bertotti et al. were able to identify HER2 gene amplification as one such mechanism of resistance to anti-EGFR therapy. © 2011 American Association for Cancer Research.


Roock W.D.,Catholic University of Leuven | Vriendt V.D.,Catholic University of Leuven | Normanno N.,Cell Biology and Biotherapy Unit | Ciardiello F.,The Second University of Naples | Tejpar S.,Catholic University of Leuven
The Lancet Oncology | Year: 2011

The discovery of mutant KRAS as a predictor of resistance to epidermal growth-factor receptor (EGFR) monoclonal antibodies brought a major change in the treatment of metastatic colorectal cancer. This seminal finding also highlighted our sparse knowledge about key signalling pathways in colorectal tumours. Drugs that inhibit oncogenic alterations such as phospho-MAP2K (also called MEK), phospho-AKT, and mutant B-RAF seem promising as single treatment or when given with EGFR inhibitors. However, our understanding of the precise role these potential drug targets have in colorectal tumours, and the oncogenic dependence that tumours might have on these components, has not progressed at the same rate. As a result, patient selection and prediction of treatment effects remain problematic. We review the role of mutations in genes other than KRAS on the efficacy of anti-EGFR therapy, and discuss strategies to target these oncogenic alterations alone or in combination with receptor tyrosine-kinase inhibition. © 2011 Elsevier Ltd.


de Luca A.,Cell Biology and Biotherapy Unit | Normanno N.,Cell Biology and Biotherapy Unit
Current Drug Targets | Year: 2010

The epidermal growth factor receptor (EGFR) and its ligands are frequently expressed in non-small-cell lung cancer (NSCLC). The EGFR tyrosine kinase inhibitors (TKIs) erlotinib and gefitinib have shown clinical activity in NSCLC. However, only a small subgroup of NSCLC patients respond to these agents, suggesting that patients' selection is critical for EGFR TKIs sensitivity. In this regard, several studies have tried to individuate prognostic and predictive factors that are associated with sensitivity or resistance to anti-EGFR agents. A strong correlation between activating mutations in the EGFR TK domain and response to erlotinib and gefitinib has been reported in different trials. However, patients without EGFR mutations might also benefit of treatment with these drugs by experiencing prolonged disease stabilization. No significant correlation between EGFR overexpression and response to treatment has been found, while controversial results have been reported regarding the association between EGFR gene amplification and clinical response to TKIs. Different mechanisms of resistance to EGFR TKIs have also been described. Mutations of KRAS, that occur in approximately 20% of NSCLC, are associated with reduced response to EGFR TKIs. The EGFR T790M mutation, that reduces the affinity of the EGFR to gefitinib and erlotinib, and MET gene amplification produce acquired resistance to anti-EGFR agents. Taken together, these findings suggest that several different molecular alterations regulate the sensitivity of NSCLC cells to EGFR TKIs, and that a comprehensive approach to this phenomenon is necessary for an appropriate selection of patients that should be treated with these drugs. © 2010 Bentham Science Publishers Ltd.


De Luca A.,Cell Biology and Biotherapy Unit | Lamura L.,Cell Biology and Biotherapy Unit | Gallo M.,Cell Biology and Biotherapy Unit | Maffia V.,Cell Biology and Biotherapy Unit | Normanno N.,Cell Biology and Biotherapy Unit
Journal of Cellular Biochemistry | Year: 2012

Several different cytokines and growth factors secreted by mesenchymal stem cells (MSCs) have been hypothesized to play a role in breast cancer progression. By using a small panel of breast cancer cell lines (MCF-7, T47D, and SK-Br-3 cells), we analyzed the role of interleukin-6 (IL-6) and vascular endothelial growth factor A (VEGF) in the cross-talk between MSCs and breast cancer cells. We performed migration assays in which breast cancer cells were allowed to migrate in response to conditioned medium from MSCs (MSCs-CM), in absence or in presence of the anti-VEGF antibody bevacizumab or an anti-IL-6 antibody, alone or in combination. We found that anti-VEGF and anti-IL-6 antibodies inhibited the migration of breast cancer cells and that the combination had an higher inhibitory effect. We next evaluated the effects of recombinant VEGF and IL-6 proteins on breast cancer cell growth and migration. IL-6 and VEGF had not significant effects on the proliferation of breast carcinoma cells. In contrast, both VEGF and IL-6 significantly increased the ability to migrate of MCF-7, T47D and SK-Br-3 cells, with the combination showing a greater effect as compared with treatment with a single protein. The combination of VEGF and IL-6 produced in breast cancer cells a more significant and more persistent activation of MAPK, AKT, and p38MAPK intracellular signaling pathways. These results suggest that MSC-secreted IL-6 and VEGF may act as paracrine factors to sustain breast cancer cell migration. J. Cell. Biochem. 113: 3363-3370, 2012. © 2012 Wiley Periodicals, Inc.


De Luca A.,Cell Biology and Biotherapy Unit | Maiello M.R.,Cell Biology and Biotherapy Unit | D'Alessio A.,Cell Biology and Biotherapy Unit | Pergameno M.,Cell Biology and Biotherapy Unit | Normanno N.,Cell Biology and Biotherapy Unit
Expert Opinion on Therapeutic Targets | Year: 2012

Introduction: The RAS/RAF/MAP kinase-ERK kinase (MEK)/extracellular-signal- regulated kinase (ERK) (MAPK) and the PI3K/AKT/mammalian target of rapamycin (mTOR) (PI3K) pathways are frequently deregulated in human cancer as a result of genetic alterations in their components or upstream activation of cell-surface receptors. These signalling cascades are regulated by complex feedback and cross-talk mechanisms. Areas covered: In this review the key components of the MAPK and AKT pathways and their molecular alterations are described. The complex interactions between these signalling cascades are also analysed. Expert opinion: The observation that the MAPK and the PI3K pathways are often deregulated in human cancer makes the components of these signalling cascades interesting targets for therapeutic intervention. Recently, the presence of compensatory loops that activate one pathway following the blockade of the other signalling cascade has been demonstrated. Therefore, the blockade of both pathways with combinations of signalling inhibitors might result in a more efficient anti-tumor effect as compared with a single agent. In addition, the MAPK and PI3K pathways are activated by mutations that coexist or can be mutually exclusive. In this regard, a large-scale characterization of the cancer genome might offer personalized cancer genomic information, which may improve the anti-tumor efficacy of signalling inhibitors. © Informa UK, Ltd.


De Luca A.,Cell Biology and Biotherapy Unit | Normanno N.,Cell Biology and Biotherapy Unit
IDrugs | Year: 2010

Tivozanib (AV-951; KRN-951), being developed by AVEO Pharmaceuticals Inc and Kyowa Hakko Kirin Co Ltd, is an orally active, ATP-competitive, small-molecule, quinoline-urea derivative that inhibits VEGFR tyrosine kinase for the potential treatment of cancer. In particular, tivozanib is able to markedly inhibit the ligand-induced phosphorylation of VEGFR-1, VEGFR-2 and VEGFR-3 at picomolar concentrations. In preclinical studies, tivozanib produced a significant inhibition of tumor growth and angiogenesis in several different xenograft tumor models in athymic rats. In a phase I clinical trial, tivozanib was safe and tolerable when administered at oral doses up to 1.5 mg on a schedule of 4 weeks on, 2 weeks off treatment. Results from a phase II clinical trial in patients with advanced renal cell carcinoma reported an overall response rate of 25.4% and a median progression-free survival of 11.8 months in patients treated with tivozanib as a single agent. Hypertension and dysphonia were the most frequent adverse events. At the time of publication, a phase III clinical trial was recruiting patients with advanced renal cancer to assess tivozanib in comparison with sorafenib. Clinical trials are currently ongoing to evaluate the safety and antitumor activity of tivozanib in breast, lung and colorectal cancer. Tivozanib might represent a promising anticancer agent in several different tumor types. © Thomson Reuters (Scientific) Ltd.

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