Detection of EGFR, KRAS and
                        BRAF mutations in metastatic cells from cerebrospinal
                    fluid

Diane Frankel; Isabelle Nanni-Metellus; Andrée Robaglia-Schlupp; Pascale Tomasini; Julien Guinde; Fabrice Barlesi; Philippe Astoul; L’Houcine Ouafik; Florent Amatore; Véronique Secq; Elise Kaspi; Patrice Roll

doi:10.1515/cclm-2017-0527

Article

Detection of EGFR, KRAS and BRAF mutations in metastatic cells from cerebrospinal fluid

Diane Frankel , Isabelle Nanni-Metellus , Andrée Robaglia-Schlupp , Pascale Tomasini , Julien Guinde , Fabrice Barlesi , Philippe Astoul , L’Houcine Ouafik , Florent Amatore , Véronique Secq , Elise Kaspi and Patrice Roll

Published/Copyright: January 8, 2018

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From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 56 Issue 5

Abstract

Background:

In lung adenocarcinoma, molecular profiling of actionable genes has become essential to set up targeted therapies. However, the feasibility and the relevance of molecular profiling from the cerebrospinal fluid (CSF) in the context of meningeal metastasis have been poorly assessed.

Methods:

We selected patients with stage IV lung adenocarcinoma harbouring metastatic cells in the CSF after cytological analysis. Seven samples from six patients were eligible for molecular testing of epidermal growth factor receptor (EGFR), V-Ki-ras2 Kirsten rat sarcoma viral oncogene homologue (KRAS), v-Raf murine sarcoma viral oncogene homologue B1 (BRAF) and human epidermal growth factor receptor 2 (HER2) mutations using quantitative polymerase chain reaction (PCR) high-resolution melting curve analysis and Sanger sequencing after DNA extraction from the cell pellets of the CSF.

Results:

Five patients showed mutations in one or two actionable genes, two harboured an EGFR mutation (exons 19 and 21), one only a KRAS mutation, one both EGFR and KRAS mutations and one a BRAF mutation. In all cases, the results of mutation testing provided new major information for patient management, leading to therapeutic adaptation. CSF molecular analysis identified mutations not detected in other neoplastic sites for two patients. In one case, the EGFR p.Thr790Met was identified. CSF was also the only sample available for genetic testing for almost all patients at the time of disease progression.

Conclusions:

When cancer cells are present in the CSF, the molecular profiling from the cell pellets is relevant, as it can detect supplemental or different mutations compared to a previous analysis of the primitive tumour or plasma cell-free DNA and allows the adaptation of the treatment strategy.

Keywords: BRAF; cerebrospinal fluid; EGFR; KRAS; lung adenocarcinoma; meningeal metastasis

Corresponding author : Patrice Roll, MD, PhD, Aix-Marseille Univ, INSERM, MMG, Faculté de Médecine de la Timone, 27 Bd Jean Moulin, 13385 Marseille Cedex 5, France, Phone: +33 491 324 939, Fax: +33 33 491 804 319

Acknowledgments

We thank Joëlle Fiteni, Nathalie Boitano and Corinne Derrien for their help and continuous support. We also thank Romain Appay for his technical assistance and Philippe Metellus for his helpful comments. We thank the “Institut National du Cancer” (INCa) for the financial support.

Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Supplemental Material:

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2017-0527).

Received: 2017-6-15

Accepted: 2017-12-4

Published Online: 2018-1-8

Published in Print: 2018-4-25

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Keywords for this article

BRAF; cerebrospinal fluid; EGFR; KRAS; lung adenocarcinoma; meningeal metastasis

Detection of EGFR, KRAS and BRAF mutations in metastatic cells from cerebrospinal fluid

Article

Abstract

Background:

Methods:

Results:

Conclusions:

Acknowledgments

References

Supplemental Material:

Supplementary Material

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