Circulating tumour cells from patients with colorectal cancer have cancer stem cell hallmarks in ex vivo culture

drug toxicity
cancer
colorectal
stem cell
rna-seq
Authors

Fanny Grillet

Elsa Bayet

Olivia Villeronce

Luke Zappia

Ebba Louise Lagerqvist

Sebastian Lunke

Emmanuelle Charafe-Jauffret

Kym Pham

Christina Molck

Nathalie Rolland

Jean François Bourgaux

Michel Prudhomme

Claire Philippe

Sophie Bravo

Jean Christophe Boyer

Lucile Canterel-Thouennon

Graham Roy Taylor

Arthur Hsu

Jean Marc Pascussi

Frédéric Hollande

Julie Pannequin

Date

July 1, 2016

Links
Citation stats
Abstract

Objective

Although counting of circulating tumour cells (CTC) has attracted a broad interest as potential markers of tumour progression and treatment response, the lack of functional characterisation of these cells had become a bottleneck in taking these observations to the clinic. Our objective was to culture these cells in order to understand them and exploit their therapeutic potential to the full.

Design

Here, hypothesising that some CTC potentially have cancer stem cell (CSC) phenotype, we generated several CTC lines from the blood of patients with advanced metastatic colorectal cancer (CRC) based on their self-renewal abilities. Multiple standard tests were then employed to characterise these cells.

Results

Our CTC lines self-renew, express CSC markers and have multilineage differentiation ability, both in vitro and in vivo. Patient-derived CTC lines are tumorigenic in subcutaneous xenografts and are also able to colonise the liver after intrasplenic injection. RNA sequencing analyses strikingly demonstrate that drug metabolising pathways represent the most upregulated feature among CTC lines in comparison with primary CRC cells grown under similar conditions. This result is corroborated by the high resistance of the CTC lines to conventional cytotoxic compounds.

Conclusions

Taken together, our results directly demonstrate the existence of patient-derived colorectal CTCs that bear all the functional attributes of CSCs. The CTC culture model described here is simple and takes <1 month from blood collection to drug testing, therefore, routine clinical application could facilitate access to personalised medicine.

Citation

BibTeX citation:
@article{grillet2016,
  author = {Grillet, Fanny and Bayet, Elsa and Villeronce, Olivia and
    Zappia, Luke and Louise Lagerqvist, Ebba and Lunke, Sebastian and
    Charafe-Jauffret, Emmanuelle and Pham, Kym and Molck, Christina and
    Rolland, Nathalie and François Bourgaux, Jean and Prudhomme, Michel
    and Philippe, Claire and Bravo, Sophie and Christophe Boyer, Jean
    and Canterel-Thouennon, Lucile and Roy Taylor, Graham and Hsu,
    Arthur and Marc Pascussi, Jean and Hollande, Frédéric and Pannequin,
    Julie},
  title = {Circulating Tumour Cells from Patients with Colorectal Cancer
    Have Cancer Stem Cell Hallmarks in Ex Vivo Culture},
  journal = {Gut},
  date = {2016-07-01},
  url = {https://lazappi.id.au/publications/2016-grillet-colorectal-CTCs/},
  doi = {10.1136/gutjnl-2016-311447},
  issn = {0017-5749, 1468-3288},
  langid = {en},
  abstract = {**Objective** Although counting of circulating tumour
    cells (CTC) has attracted a broad interest as potential markers of
    tumour progression and treatment response, the lack of functional
    characterisation of these cells had become a bottleneck in taking
    these observations to the clinic. Our objective was to culture these
    cells in order to understand them and exploit their therapeutic
    potential to the full. **Design** Here, hypothesising that some CTC
    potentially have cancer stem cell (CSC) phenotype, we generated
    several CTC lines from the blood of patients with advanced
    metastatic colorectal cancer (CRC) based on their self-renewal
    abilities. Multiple standard tests were then employed to
    characterise these cells. **Results** Our CTC lines self-renew,
    express CSC markers and have multilineage differentiation ability,
    both in vitro and in vivo. Patient-derived CTC lines are tumorigenic
    in subcutaneous xenografts and are also able to colonise the liver
    after intrasplenic injection. RNA sequencing analyses strikingly
    demonstrate that drug metabolising pathways represent the most
    upregulated feature among CTC lines in comparison with primary CRC
    cells grown under similar conditions. This result is corroborated by
    the high resistance of the CTC lines to conventional cytotoxic
    compounds. **Conclusions** Taken together, our results directly
    demonstrate the existence of patient-derived colorectal CTCs that
    bear all the functional attributes of CSCs. The CTC culture model
    described here is simple and takes \textless1 month from blood
    collection to drug testing, therefore, routine clinical application
    could facilitate access to personalised medicine.}
}
For attribution, please cite this work as:
Grillet, Fanny, Elsa Bayet, Olivia Villeronce, Luke Zappia, Ebba Louise Lagerqvist, Sebastian Lunke, Emmanuelle Charafe-Jauffret, et al. 2016. “Circulating Tumour Cells from Patients with Colorectal Cancer Have Cancer Stem Cell Hallmarks in Ex Vivo Culture.” Gut, July. https://doi.org/10.1136/gutjnl-2016-311447.