PMID- 32303258
OWN - NLM
STAT- MEDLINE
DCOM- 20210531
LR  - 20210531
IS  - 2051-5960 (Electronic)
IS  - 2051-5960 (Linking)
VI  - 8
IP  - 1
DP  - 2020 Apr 17
TI  - Simultaneous detection of EGFR amplification and EGFRvIII variant using digital 
      PCR-based method in glioblastoma.
PG  - 52
LID - 10.1186/s40478-020-00917-6 [doi]
LID - 52
AB  - Epidermal growth factor receptor (EGFR) amplification and EGFR variant III 
      (EGFRvIII, deletion of exons 2-7) are of clinical interest for glioblastoma. The 
      aim was to develop a digital PCR (dPCR)-based method using locked nucleic acid 
      (LNA)-based hydrolysis probes, allowing the simultaneous detection of the EGFR 
      amplification and EGFRvIII variant. Sixty-two patients were included. An 
      exploratory cohort (n = 19) was used to develop the dPCR assay using three 
      selected amplicons within the EGFR gene, targeting intron 1 (EGFR1), junction of 
      exon 3 and intron 3 (EGFR2) and intron 22 (EGFR3). The copy number of EGFR was 
      estimated by the relative quantification of EGFR1, EGFR2 and EGFR3 amplicon 
      droplets compared to the droplets of a reference gene. EGFRvIII was identified by 
      comparing the copy number of the EGFR2 amplicon to either the EGFR1 or EGFR3 
      amplicon. dPCR results were compared to fluorescence in situ hybridization (FISH) 
      and next-generation sequencing for amplification; and to RT-PCR-based method for 
      EGFRvIII. The dPCR assay was then tested in a validation cohort (n = 43). A total 
      of 8/19 EGFR-amplified and 5/19 EGFRvIII-positive tumors were identified in the 
      exploratory cohort. Compared to FISH, the EGFR3 dPCR assay detected all 
      EGFR-amplified tumors (8/8, 100%) and had the highest concordance with the copy 
      number estimation by NGS. The concordance between RT-PCR and dPCR was also 100% 
      for detecting EGFRvIII using an absolute difference of 10.8 for the copy number 
      between EGFR2 and EGFR3 probes. In the validation cohort, the sensitivity and 
      specificity of dPCR using EGFR3 probes were 100% for the EGFR amplification 
      detection compared to FISH (19/19). EGFRvIII was detected by dPCR in 8 
      EGFR-amplified patients and confirmed by RT-PCR. Compared to FISH, the 
      EGFR2/EGFR3 dPCR assay was estimated with a one-half cost value. These results 
      highlight that dPCR allowed the simultaneous detection of EGFR amplification and 
      EGFRvIII for glioblastoma.
FAU - Fontanilles, Maxime
AU  - Fontanilles M
AD  - Inserm U1245, Normandie Univ, UNIROUEN, IRON group, Normandy Centre for Genomic 
      and Personalized Medicine, Rouen University Hospital, F-76031, Rouen, France. 
      maxime.fontanilles@chb.unicancer.fr.
AD  - Department of Medical Oncology, Cancer Centre Henri Becquerel, Rue d'Amiens, 
      F-76038, Rouen, France. maxime.fontanilles@chb.unicancer.fr.
FAU - Marguet, Florent
AU  - Marguet F
AD  - Inserm U1245, Normandie Univ, UNIROUEN, Normandy Centre for Genomic and 
      Personalized Medicine, Rouen University Hospital, F-76031, Rouen, France.
AD  - Department of Pathology, Rouen University Hospital, F-76031, Rouen, France.
FAU - Ruminy, Philippe
AU  - Ruminy P
AD  - Inserm U1245, Normandie Univ, UNIROUEN, Normandy Centre for Genomic and 
      Personalized Medicine, Rouen University Hospital, F-76031, Rouen, France.
FAU - Basset, Carole
AU  - Basset C
AD  - Department of Pathology, Rouen University Hospital, F-76031, Rouen, France.
FAU - Noel, Adrien
AU  - Noel A
AD  - Inserm U1245, Normandie Univ, UNIROUEN, IRON group, Normandy Centre for Genomic 
      and Personalized Medicine, Rouen University Hospital, F-76031, Rouen, France.
FAU - Beaussire, Ludivine
AU  - Beaussire L
AD  - Inserm U1245, Normandie Univ, UNIROUEN, IRON group, Normandy Centre for Genomic 
      and Personalized Medicine, Rouen University Hospital, F-76031, Rouen, France.
FAU - Viennot, Mathieu
AU  - Viennot M
AD  - Inserm U1245, Normandie Univ, UNIROUEN, Normandy Centre for Genomic and 
      Personalized Medicine, Rouen University Hospital, F-76031, Rouen, France.
FAU - Viailly, Pierre-Julien
AU  - Viailly PJ
AD  - Department of Pathology, Rouen University Hospital, F-76031, Rouen, France.
FAU - Cassinari, Kevin
AU  - Cassinari K
AD  - Department of Genetics, Inserm U1245, Normandie Univ, UNIROUEN, Normandy Centre 
      for Genomic and Personalized Medicine, Rouen University Hospital, F-76031, Rouen, 
      France.
FAU - Chambon, Pascal
AU  - Chambon P
AD  - Department of Genetics, Inserm U1245, Normandie Univ, UNIROUEN, Normandy Centre 
      for Genomic and Personalized Medicine, Rouen University Hospital, F-76031, Rouen, 
      France.
FAU - Richard, Doriane
AU  - Richard D
AD  - Department of Statistics and Clinical Research Unit, Cancer Centre Henri 
      Becquerel, Rouen, F-76038, France.
FAU - Alexandru, Cristina
AU  - Alexandru C
AD  - Department of Medical Oncology, Cancer Centre Henri Becquerel, Rue d'Amiens, 
      F-76038, Rouen, France.
FAU - Tennevet, Isabelle
AU  - Tennevet I
AD  - Department of Medical Oncology, Cancer Centre Henri Becquerel, Rue d'Amiens, 
      F-76038, Rouen, France.
FAU - Langlois, Olivier
AU  - Langlois O
AD  - Department of Neurosurgery, Rouen University Hospital, F-76031, Rouen, France.
FAU - Di Fiore, Frederic
AU  - Di Fiore F
AD  - Inserm U1245, Normandie Univ, UNIROUEN, IRON group, Normandy Centre for Genomic 
      and Personalized Medicine, Rouen University Hospital, F-76031, Rouen, France.
AD  - Department of Medical Oncology, Cancer Centre Henri Becquerel, Rue d'Amiens, 
      F-76038, Rouen, France.
AD  - Department of Gastroenterology, Rouen University Hospital, F-76031, Rouen, 
      France.
FAU - Laquerriere, Annie
AU  - Laquerriere A
AD  - Inserm U1245, Normandie Univ, UNIROUEN, Normandy Centre for Genomic and 
      Personalized Medicine, Rouen University Hospital, F-76031, Rouen, France.
AD  - Department of Pathology, Rouen University Hospital, F-76031, Rouen, France.
FAU - Clatot, Florian
AU  - Clatot F
AD  - Inserm U1245, Normandie Univ, UNIROUEN, IRON group, Normandy Centre for Genomic 
      and Personalized Medicine, Rouen University Hospital, F-76031, Rouen, France.
AD  - Department of Medical Oncology, Cancer Centre Henri Becquerel, Rue d'Amiens, 
      F-76038, Rouen, France.
FAU - Sarafan-Vasseur, Nasrin
AU  - Sarafan-Vasseur N
AD  - Inserm U1245, Normandie Univ, UNIROUEN, IRON group, Normandy Centre for Genomic 
      and Personalized Medicine, Rouen University Hospital, F-76031, Rouen, France.
LA  - eng
PT  - Clinical Trial
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200417
PL  - England
TA  - Acta Neuropathol Commun
JT  - Acta neuropathologica communications
JID - 101610673
RN  - 0 (Biomarkers)
RN  - EC 2.7.10.1 (ErbB Receptors)
RN  - YF1K15M17Y (Temozolomide)
SB  - IM
MH  - Adult
MH  - Aged
MH  - Biomarkers/analysis
MH  - Brain Neoplasms/*genetics/therapy
MH  - Chemoradiotherapy/adverse effects
MH  - ErbB Receptors
MH  - Female
MH  - Gene Amplification
MH  - Glioblastoma/*genetics/therapy
MH  - Humans
MH  - Male
MH  - Middle Aged
MH  - Polymerase Chain Reaction/*methods
MH  - Temozolomide/adverse effects
PMC - PMC7165387
OTO - NOTNLM
OT  - Cost-effectiveness
OT  - Digital PCR
OT  - EGFR amplification
OT  - EGFRvIII variant
OT  - Glioblastoma
COIS- Maxime Fontanilles: Reports Honoria from Bristol-Myers Squibb(R) and Congress Fee 
      from La Roche-Hauffman(R).
EDAT- 2020/04/19 06:00
MHDA- 2021/06/01 06:00
PMCR- 2020/04/17
CRDT- 2020/04/19 06:00
PHST- 2020/01/29 00:00 [received]
PHST- 2020/03/13 00:00 [accepted]
PHST- 2020/04/19 06:00 [entrez]
PHST- 2020/04/19 06:00 [pubmed]
PHST- 2021/06/01 06:00 [medline]
PHST- 2020/04/17 00:00 [pmc-release]
AID - 10.1186/s40478-020-00917-6 [pii]
AID - 917 [pii]
AID - 10.1186/s40478-020-00917-6 [doi]
PST - epublish
SO  - Acta Neuropathol Commun. 2020 Apr 17;8(1):52. doi: 10.1186/s40478-020-00917-6.