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

Giuseppe Roscilli, Claudia De Vitis, Fabiana Fosca Ferrara, Alessia Noto, Emanuela Cherubini, Alberto Ricci, Salvatore Mariotta, Enrico Giarnieri, Maria Rosaria Giovagnoli, Maria Rosaria Torrisi, Francesca Bergantino, Susan Costantini, Francesca Fenizia, Matilde Lambiase, Luigi Aurisicchio, Nicola Normanno, Gennaro Ciliberto, Rita Mancini

Research output: Contribution to journalArticle

Abstract

Background: 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. Methods: 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. Results: 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. Conclusions: Results obtained using primary cultured cells from MPEs underscore the heterogeneity of NSCLC in advanced stage as indicated by drug response and mutation profile. Comparison of data obtained from in vitro assays with patients' responses to therapy leads to the conclusion that this strategy may provide a potentially useful approach for evaluating individual chemosensitivity profile and tailor the therapy accordingly. Furthermore, combining MPE-derived primary cultures with their genomic testing allows to identify patients eligible to trials with novel targeted agents.

Original languageEnglish
Article number61
JournalJournal of Translational Medicine
Volume14
Issue number1
DOIs
Publication statusPublished - Feb 29 2016

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Malignant Pleural Effusion
Cell culture
Cell Culture Techniques
Cells
Chemotherapy
Pharmaceutical Preparations
Non-Small Cell Lung Carcinoma
Tumors
Assays
Neoplasms
Centrifugation
Drug Therapy
Mutation
Lung Neoplasms
Heterografts
Therapeutics
Interleukin-2
Adenocarcinoma of lung
Density Gradient Centrifugation
Knockout Mice

Keywords

  • In vitro chemosensitivity
  • Malignant pleural effusions
  • Next generation sequencing
  • NSCLC primary cultures
  • PDX

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

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

In: Journal of Translational Medicine, Vol. 14, No. 1, 61, 29.02.2016.

Research output: Contribution to journalArticle

Roscilli, G, De Vitis, C, Ferrara, FF, Noto, A, Cherubini, E, Ricci, A, Mariotta, S, Giarnieri, E, Giovagnoli, MR, Torrisi, MR, Bergantino, F, Costantini, S, Fenizia, F, Lambiase, M, Aurisicchio, L, Normanno, N, Ciliberto, G & Mancini, R 2016, 'Human lung adenocarcinoma cell cultures derived from malignant pleural effusions as model system to predict patients chemosensitivity', Journal of Translational Medicine, vol. 14, no. 1, 61. https://doi.org/10.1186/s12967-016-0816-x
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. / Human lung adenocarcinoma cell cultures derived from malignant pleural effusions as model system to predict patients chemosensitivity. In: Journal of Translational Medicine. 2016 ; Vol. 14, No. 1.
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T1 - Human lung adenocarcinoma cell cultures derived from malignant pleural effusions as model system to predict patients chemosensitivity

AU - Roscilli, Giuseppe

AU - De Vitis, Claudia

AU - Ferrara, Fabiana Fosca

AU - Noto, Alessia

AU - Cherubini, Emanuela

AU - Ricci, Alberto

AU - Mariotta, Salvatore

AU - Giarnieri, Enrico

AU - Giovagnoli, Maria Rosaria

AU - Torrisi, Maria Rosaria

AU - Bergantino, Francesca

AU - Costantini, Susan

AU - Fenizia, Francesca

AU - Lambiase, Matilde

AU - Aurisicchio, Luigi

AU - Normanno, Nicola

AU - Ciliberto, Gennaro

AU - Mancini, Rita

PY - 2016/2/29

Y1 - 2016/2/29

N2 - Background: 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. Methods: 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. Results: 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. Conclusions: Results obtained using primary cultured cells from MPEs underscore the heterogeneity of NSCLC in advanced stage as indicated by drug response and mutation profile. Comparison of data obtained from in vitro assays with patients' responses to therapy leads to the conclusion that this strategy may provide a potentially useful approach for evaluating individual chemosensitivity profile and tailor the therapy accordingly. Furthermore, combining MPE-derived primary cultures with their genomic testing allows to identify patients eligible to trials with novel targeted agents.

AB - Background: 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. Methods: 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. Results: 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. Conclusions: Results obtained using primary cultured cells from MPEs underscore the heterogeneity of NSCLC in advanced stage as indicated by drug response and mutation profile. Comparison of data obtained from in vitro assays with patients' responses to therapy leads to the conclusion that this strategy may provide a potentially useful approach for evaluating individual chemosensitivity profile and tailor the therapy accordingly. Furthermore, combining MPE-derived primary cultures with their genomic testing allows to identify patients eligible to trials with novel targeted agents.

KW - In vitro chemosensitivity

KW - Malignant pleural effusions

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