Conditionally reprogrammed cells (CRC) methodology does not allow the in vitro expansion of patient-derived primary and metastatic lung cancer cells

G. Sette, V. Salvati, I. Giordani, E. Pilozzi, D. Quacquarini, E. Duranti, F. De Nicola, M. Pallocca, M. Fanciulli, M. Falchi, R. Pallini, R. De Maria, A. Eramo

Research output: Contribution to journalArticle

Abstract

Availability of tumor and non-tumor patient-derived models would promote the development of more effective therapeutics for non-small cell lung cancer (NSCLC). Recently, conditionally reprogrammed cells (CRC) methodology demonstrated exceptional potential for the expansion of epithelial cells from patient tissues. However, the possibility to expand patient-derived lung cancer cells using CRC protocols is controversial. Here, we used CRC approach to expand cells from non-tumoral and tumor biopsies of patients with primary or metastatic NSCLC as well as pulmonary metastases of colorectal or breast cancers. CRC cultures were obtained from both tumor and non-malignant tissues with extraordinary high efficiency. Tumor cells were tracked in vitro through tumorigenicity assay, monitoring of tumor-specific genetic alterations and marker expression. Cultures were composed of EpCAM+ lung epithelial cells lacking tumorigenic potential. NSCLC biopsies-derived cultures rapidly lost patient-specific genetic mutations or tumor antigens. Similarly, pulmonary metastases of colon or breast cancer generated CRC cultures of lung epithelial cells. All CRC cultures examined displayed epithelial lung stem cell phenotype and function. In contrast, brain metastatic lung cancer biopsies failed to generate CRC cultures. In conclusion, patient-derived primary and metastatic lung cancer cells were negatively selected under CRC conditions, limiting the expansion to non-malignant lung epithelial stem cells from either tumor or non-tumor tissue sources. Thus, CRC approach cannot be applied for direct therapeutic testing of patient lung tumor cells, as the tumor-derived CRC cultures are composed of (non-tumoral) airway basal cells. © 2018 UICC
Original languageEnglish
Pages (from-to)88-99
Number of pages12
JournalInternational Journal of Cancer
Volume143
Issue number1
DOIs
Publication statusPublished - 2018

Fingerprint

Lung Neoplasms
Lung
Cell Culture Techniques
Neoplasms
Epithelial Cells
Non-Small Cell Lung Carcinoma
Biopsy
In Vitro Techniques
Stem Cells
Breast Neoplasms
Neoplasm Metastasis
Neoplasm Antigens
Genetic Markers
Colonic Neoplasms
Colorectal Neoplasms
Phenotype
Mutation
Brain
Therapeutics

Keywords

  • anaplastic lymphoma kinase
  • CD56 antigen
  • chromogranin A
  • cytokeratin 20
  • cytokeratin 5
  • cytokeratin 6
  • cytokeratin 7
  • cytokeratin AE1
  • cytokeratin AE3
  • epidermal growth factor receptor
  • epidermal growth factor receptor 2
  • estrogen receptor
  • homeobox protein Nkx 2.1
  • napsin A
  • progesterone receptor
  • protein p63
  • ROS1 protein
  • synaptophysin
  • transcription factor Cdx2
  • tumor marker
  • unclassified drug
  • tumor marker, adult
  • aged
  • animal cell
  • animal experiment
  • animal model
  • Article
  • assay
  • breast cancer
  • cancer cell culture
  • cell culture technique
  • cell expansion
  • clinical article
  • colorectal cancer
  • conditionally reprogrammed cell culture
  • controlled study
  • epithelial lung stem cell
  • female
  • gene expression
  • gene mutation
  • human
  • human cell
  • human tissue
  • in vitro study
  • lung alveolus epithelium cell
  • lung cancer cell line
  • lung metastasis
  • male
  • middle aged
  • mouse
  • non small cell lung cancer
  • nonhuman
  • priority journal
  • stem cell
  • tumor biopsy
  • tumorigenicity assay
  • animal
  • biopsy
  • breast tumor
  • cancer transplantation
  • cell reprogramming technique
  • colon tumor
  • cytology
  • epithelium cell
  • genetics
  • lung tumor
  • mutation
  • pathology
  • phenotype
  • procedures
  • secondary
  • tumor cell culture, Aged
  • Animals
  • Biomarkers, Tumor
  • Biopsy
  • Breast Neoplasms
  • Carcinoma, Non-Small-Cell Lung
  • Cellular Reprogramming Techniques
  • Colonic Neoplasms
  • Epithelial Cells
  • Female
  • Humans
  • Lung Neoplasms
  • Male
  • Mice
  • Middle Aged
  • Mutation
  • Neoplasm Transplantation
  • Phenotype
  • Stem Cells
  • Tumor Cells, Cultured

Cite this

Conditionally reprogrammed cells (CRC) methodology does not allow the in vitro expansion of patient-derived primary and metastatic lung cancer cells. / Sette, G.; Salvati, V.; Giordani, I.; Pilozzi, E.; Quacquarini, D.; Duranti, E.; De Nicola, F.; Pallocca, M.; Fanciulli, M.; Falchi, M.; Pallini, R.; De Maria, R.; Eramo, A.

In: International Journal of Cancer, Vol. 143, No. 1, 2018, p. 88-99.

Research output: Contribution to journalArticle

Sette, G, Salvati, V, Giordani, I, Pilozzi, E, Quacquarini, D, Duranti, E, De Nicola, F, Pallocca, M, Fanciulli, M, Falchi, M, Pallini, R, De Maria, R & Eramo, A 2018, 'Conditionally reprogrammed cells (CRC) methodology does not allow the in vitro expansion of patient-derived primary and metastatic lung cancer cells', International Journal of Cancer, vol. 143, no. 1, pp. 88-99. https://doi.org/10.1002/ijc.31260
Sette, G. ; Salvati, V. ; Giordani, I. ; Pilozzi, E. ; Quacquarini, D. ; Duranti, E. ; De Nicola, F. ; Pallocca, M. ; Fanciulli, M. ; Falchi, M. ; Pallini, R. ; De Maria, R. ; Eramo, A. / Conditionally reprogrammed cells (CRC) methodology does not allow the in vitro expansion of patient-derived primary and metastatic lung cancer cells. In: International Journal of Cancer. 2018 ; Vol. 143, No. 1. pp. 88-99.
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AU - Sette, G.

AU - Salvati, V.

AU - Giordani, I.

AU - Pilozzi, E.

AU - Quacquarini, D.

AU - Duranti, E.

AU - De Nicola, F.

AU - Pallocca, M.

AU - Fanciulli, M.

AU - Falchi, M.

AU - Pallini, R.

AU - De Maria, R.

AU - Eramo, A.

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N2 - Availability of tumor and non-tumor patient-derived models would promote the development of more effective therapeutics for non-small cell lung cancer (NSCLC). Recently, conditionally reprogrammed cells (CRC) methodology demonstrated exceptional potential for the expansion of epithelial cells from patient tissues. However, the possibility to expand patient-derived lung cancer cells using CRC protocols is controversial. Here, we used CRC approach to expand cells from non-tumoral and tumor biopsies of patients with primary or metastatic NSCLC as well as pulmonary metastases of colorectal or breast cancers. CRC cultures were obtained from both tumor and non-malignant tissues with extraordinary high efficiency. Tumor cells were tracked in vitro through tumorigenicity assay, monitoring of tumor-specific genetic alterations and marker expression. Cultures were composed of EpCAM+ lung epithelial cells lacking tumorigenic potential. NSCLC biopsies-derived cultures rapidly lost patient-specific genetic mutations or tumor antigens. Similarly, pulmonary metastases of colon or breast cancer generated CRC cultures of lung epithelial cells. All CRC cultures examined displayed epithelial lung stem cell phenotype and function. In contrast, brain metastatic lung cancer biopsies failed to generate CRC cultures. In conclusion, patient-derived primary and metastatic lung cancer cells were negatively selected under CRC conditions, limiting the expansion to non-malignant lung epithelial stem cells from either tumor or non-tumor tissue sources. Thus, CRC approach cannot be applied for direct therapeutic testing of patient lung tumor cells, as the tumor-derived CRC cultures are composed of (non-tumoral) airway basal cells. © 2018 UICC

AB - Availability of tumor and non-tumor patient-derived models would promote the development of more effective therapeutics for non-small cell lung cancer (NSCLC). Recently, conditionally reprogrammed cells (CRC) methodology demonstrated exceptional potential for the expansion of epithelial cells from patient tissues. However, the possibility to expand patient-derived lung cancer cells using CRC protocols is controversial. Here, we used CRC approach to expand cells from non-tumoral and tumor biopsies of patients with primary or metastatic NSCLC as well as pulmonary metastases of colorectal or breast cancers. CRC cultures were obtained from both tumor and non-malignant tissues with extraordinary high efficiency. Tumor cells were tracked in vitro through tumorigenicity assay, monitoring of tumor-specific genetic alterations and marker expression. Cultures were composed of EpCAM+ lung epithelial cells lacking tumorigenic potential. NSCLC biopsies-derived cultures rapidly lost patient-specific genetic mutations or tumor antigens. Similarly, pulmonary metastases of colon or breast cancer generated CRC cultures of lung epithelial cells. All CRC cultures examined displayed epithelial lung stem cell phenotype and function. In contrast, brain metastatic lung cancer biopsies failed to generate CRC cultures. In conclusion, patient-derived primary and metastatic lung cancer cells were negatively selected under CRC conditions, limiting the expansion to non-malignant lung epithelial stem cells from either tumor or non-tumor tissue sources. Thus, CRC approach cannot be applied for direct therapeutic testing of patient lung tumor cells, as the tumor-derived CRC cultures are composed of (non-tumoral) airway basal cells. © 2018 UICC

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KW - chromogranin A

KW - cytokeratin 20

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KW - cytokeratin AE1

KW - cytokeratin AE3

KW - epidermal growth factor receptor

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KW - homeobox protein Nkx 2.1

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KW - ROS1 protein

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KW - unclassified drug

KW - tumor marker, adult

KW - aged

KW - animal cell

KW - animal experiment

KW - animal model

KW - Article

KW - assay

KW - breast cancer

KW - cancer cell culture

KW - cell culture technique

KW - cell expansion

KW - clinical article

KW - colorectal cancer

KW - conditionally reprogrammed cell culture

KW - controlled study

KW - epithelial lung stem cell

KW - female

KW - gene expression

KW - gene mutation

KW - human

KW - human cell

KW - human tissue

KW - in vitro study

KW - lung alveolus epithelium cell

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KW - mouse

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KW - priority journal

KW - stem cell

KW - tumor biopsy

KW - tumorigenicity assay

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KW - biopsy

KW - breast tumor

KW - cancer transplantation

KW - cell reprogramming technique

KW - colon tumor

KW - cytology

KW - epithelium cell

KW - genetics

KW - lung tumor

KW - mutation

KW - pathology

KW - phenotype

KW - procedures

KW - secondary

KW - tumor cell culture, Aged

KW - Animals

KW - Biomarkers, Tumor

KW - Biopsy

KW - Breast Neoplasms

KW - Carcinoma, Non-Small-Cell Lung

KW - Cellular Reprogramming Techniques

KW - Colonic Neoplasms

KW - Epithelial Cells

KW - Female

KW - Humans

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