Real-time molecular optical micro-imaging of EGFR mutations using a fluorescent erlotinib based tracer

Maxime Patout (1) , Florian Guisier (2) , Xavier Brune (3) , Pierre Bohn (4) , Anthony Romieu (3) , Nasrin Sarafan-Vasseur (5, 6) , Richard Sesboüé (5) , Pierre-Yves Renard (3) , Luc Thiberville (7) , Mathieu Salaün (1)

1 Service de pneumologie, oncologie thoracique et soins intensifs respiratoires [Rouen]
2 CHU Rouen
3 COBRA - Chimie Organique et Bioorganique : Réactivité et Analyse
4 QuantIF-LITIS - Equipe Quantification en Imagerie Fonctionnelle
5 GMFC - Génétique du cancer et des maladies neuropsychiatriques
6 GPMCND - Génomique et Médecine Personnalisée du Cancer et des Maladies Neuropsychiatriques
7 LITIS - Laboratoire d'Informatique, de Traitement de l'Information et des Systèmes

Background EGFR mutations are routinely explored in lung adenocarcinoma by sequencing tumoral DNA. The aim of this study was to evaluate a fluorescent-labelled erlotinib based theranostic agent for the molecular imaging of mutated EGFR tumours in vitro and ex vivo using a mice xenograft model and fibred confocal fluorescence microscopy (FCFM). Methods The fluorescent tracer was synthesized in our laboratory by addition of fluorescein to an erlotinib molecule. Three human adenocarcinoma cell lines with mutated EGFR (HCC827, H1975 and H1650) and one with wild-type EGFR (A549) were xenografted on 35 Nude mice. MTT viability assay was performed after exposure to our tracer. In vitro imaging was performed at 1 μM tracer solution, and ex vivo imaging was performed on fresh tumours excised from mice and exposed to a 1 μM tracer solution in PBS for 1 h. Real-time molecular imaging was performed using FCFM and median fluorescence intensity (MFI) was recorded for each experiment. Results MTT viability assay confirmed that addition of fluorescein to erlotinib did not suppress the cytotoxic of erlotinib on tumoral cells. In vitro FCFM imaging showed that our tracer was able to distinguish cell lines with mutated EGFR from those lines with wild-type EGFR (p < 0.001). Ex vivo FCFM imaging of xenografts with mutated EGFR had a significantly higher MFI than wild-type (p < 0.001). At a cut-off value of 354 Arbitrary Units, MFI of our tracer had a sensitivity of 100% and a specificity of 96.3% for identifying mutated EGFR tumours. Conclusion Real time molecular imaging using fluorescent erlotinib is able to identify ex vivo tumours with EGFR mutations.

Domains: Life Sciences [q-bio]

Maxime Patout, Florian Guisier, Xavier Brune, Pierre Bohn, Anthony Romieu, et al.. Real-time molecular optical micro-imaging of EGFR mutations using a fluorescent erlotinib based tracer. BMC Pulmonary Medicine, 2019, 19, pp.3. ⟨10.1186/s12890-018-0760-z⟩. ⟨hal-02353026⟩