Human lung adenocarcinoma cell cultures derived from malignant pleural effusions as model system to predict patients chemosensitivity

• Published in: Journal of translational medicine 2016-02, Vol.14 (61), p.61-61, Article 61
• Main Authors: Roscilli, Giuseppe, De Vitis, Claudia, Ferrara, Fabiana Fosca, Noto, Alessia, Cherubini, Emanuela, Ricci, Alberto, Mariotta, Salvatore, Giarnieri, Enrico, Giovagnoli, Maria Rosaria, Torrisi, Maria Rosaria, Bergantino, Francesca, Costantini, Susan, Fenizia, Francesca, Lambiase, Matilde, Aurisicchio, Luigi, Normanno, Nicola, Ciliberto, Gennaro, Mancini, Rita
• Format: Article
• Language: English
• Subjects: Adenocarcinoma; Adenocarcinoma - complications; Adenocarcinoma - drug therapy; Adenocarcinoma - genetics; Adenocarcinoma of Lung; Aged; Antineoplastic Agents - pharmacology; Antineoplastic Agents - therapeutic use; Biological Assay; Cancer; Care and treatment; Cell Proliferation - drug effects; Chemotherapy; Complications and side effects; Development and progression; DNA Mutational Analysis; Erlotinib Hydrochloride - pharmacology; Erlotinib Hydrochloride - therapeutic use; Exome - genetics; Female; Genetic Heterogeneity; Health aspects; Humans; Lung cancer, Non-small cell; Lung Neoplasms - complications; Lung Neoplasms - drug therapy; Lung Neoplasms - genetics; Male; Metabolic Networks and Pathways - drug effects; Models, Biological; Mutation - genetics; Patient outcomes; Pleural Effusion, Malignant - complications; Pleural Effusion, Malignant - drug therapy; Pleural Effusion, Malignant - genetics; Pleural Effusion, Malignant - pathology; Signal Transduction - drug effects; Tumor Cells, Cultured; Xenograft Model Antitumor Assays
• ISSN: 1479-5876; 1479-5876
• DOI: 10.1186/s12967-016-0816-x

Lung cancer is the leading cause of cancer related deaths and Malignant Pleural Effusion (MPE) is a frequent complication. Current therapies suffer from lack of efficacy in a great percentage of cases, especially when cancer is diagnosed at a late stage. Moreover patients' responses vary and the outcome is unpredictable. Therefore, the identification of patients who will benefit most of chemotherapy treatment is important for accurate prognostication and better outcome. In this study, using malignant pleural effusions (MPE) from non-small cell lung cancer (NSCLC) patients, we established a collection of patient-derived Adenocarcinoma cultures which were characterized for their sensitivity to chemotherapeutic drugs used in the clinical practice. Tumor cells present in MPEs of patients with NSCLC were isolated by density gradient centrifugation, placed in culture and genotyped by next generation sequencing. In a subset of cases patient derived xenografts (PDX) were obtained upon tumor cell inoculation in rag2/IL2 knock-out mice. Isolated primary cultures were characterized and tested for drug sensitivity by in vitro proliferation assays. Additivity, antagonism or synergy for combinatorial treatments were determined by analysis with the Calcusyn software. We have optimized isolation procedures and culture conditions to expand in vitro primary cultures from Malignant Pleural Effusions (MPEs) of patients affected by lung adenocarcinomas, the most frequent form of non small cell lung cancer. Using this approach we have been able to establish 16 primary cultures from MPEs. Cells were banked at low passages and were characterized for their mutational pattern by next generation sequencing for most common driver mutations in lung cancer. Moreover, amplified cultures were shown to engraft with high efficiency when injected in immunocompromised mice. Cancer cell sensitivity to drugs used in standard chemotherapy regimens was assessed either individually or in combination. Differential chemosensitivity and different mutation profiles were observed which suggests that this isolation method could provide a platform for predicting the efficacy of chemotherapy in the clinical setting. Most importantly for six patients it was possible to establish a correlation between drug response in vitro and response to therapy in the clinic. Results obtained using primary cultured cells from MPEs underscore the heterogeneity of NSCLC in advanced stage as indicated by drug response and