title: Quantification of EVI1 transcript levels in acute myeloid leukemia by RT-qPCR analysis: A study by the ALFA Group.
abstract: EVI1 overexpression confers poor prognosis in acute myeloid leukemia (AML). Quantification of EVI1 expression has been mainly assessed by real-time quantitative PCR (RT-qPCR) based on relative quantification of EVI1-1D splice variant. In this study, we developed a RT-qPCR assay to perform quantification of EVI1 expression covering the different splice variants. A sequence localized in EVI1 exons 14 and 15 was cloned into plasmids that were used to establish RT-qPCR standard curves. Threshold values to define EVI1 overexpression were determined using 17 bone marrow (BM) and 31 peripheral blood (PB) control samples and were set at 1% in BM and 0.5% in PB. Samples from 64 AML patients overexpressing EVI1 included in the ALFA-0701 or -0702 trials were collected at diagnosis and during follow-up (n=152). Median EVI1 expression at AML diagnosis was 23.3% in BM and 3.6% in PB. EVI1 expression levels significantly decreased between diagnostic and post-induction samples, with an average variation from 21.6% to 3.56% in BM and from 4.0% to 0.22% in PB, but did not exceed 1 log10 reduction. Our study demonstrates that the magnitude of reduction in EVI1 expression levels between AML diagnosis and follow-up is not sufficient to allow sensitive detection of minimal residual disease.
authors.affiliation: Hematology Laboratory, Biology and Pathology Center, CHRU of Lille, France; University of Lille Nord de France, Lille, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Smol
authors.affiliation: Hematology Laboratory, Biology and Pathology Center, CHRU of Lille, France; University of Lille Nord de France, Lille, France; UMR-S 1172, Team 3, Cancer Research Institute of Lille, Lille, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Nibourel
authors.affiliation: Hematology Laboratory, Biology and Pathology Center, CHRU of Lille, France; University of Lille Nord de France, Lille, France; UMR-S 1172, Team 3, Cancer Research Institute of Lille, Lille, France.
authors.forename: Alice
authors.initials: A
authors.lastname: Marceau-Renaut
authors.affiliation: ALFA coordination, Saint-Louis Hospital, Paris, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Celli-Lebras
authors.affiliation: University of Lille Nord de France, Lille, France; UMR-S 1172, Team 3, Cancer Research Institute of Lille, Lille, France; Hematology Department, CHRU of Lille, Lille, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Berthon
authors.affiliation: University of Lille Nord de France, Lille, France; UMR-S 1172, Team 3, Cancer Research Institute of Lille, Lille, France; Hematology Department, CHRU of Lille, Lille, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Quesnel
authors.affiliation: Hematology Department, Saint-Louis Hospital, APHP, Paris, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Boissel
authors.affiliation: Cytogenetics Laboratory, Versailles Hospital, Le Chesnay, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Terré
authors.affiliation: Hematology Department, Edouard Herriot Hospital, Lyon, France.
authors.forename: Xavier
authors.initials: X
authors.lastname: Thomas
authors.affiliation: Hematology Department, Versailles Hospital, Le Chesnay, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Castaigne
authors.affiliation: Hematology Department, Saint-Louis Hospital, APHP, Paris, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Dombret
authors.affiliation: Hematology Laboratory, Biology and Pathology Center, CHRU of Lille, France; University of Lille Nord de France, Lille, France; UMR-S 1172, Team 3, Cancer Research Institute of Lille, Lille, France.
authors.forename: Claude
authors.initials: C
authors.lastname: Preudhomme
authors.affiliation: Hematology Laboratory, Biology and Pathology Center, CHRU of Lille, France; University of Lille Nord de France, Lille, France; UMR-S 1172, Team 3, Cancer Research Institute of Lille, Lille, France. Electronic address: aline.renneville@chru-lille.fr.
authors.forename: Aline
authors.initials: A
authors.lastname: Renneville
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001773:Blood Cells / Q000378:metabolism; D001854:Bone Marrow Cells / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000097:blood / Q000235:genetics; D005260:Female; D005500:Follow-Up Studies; D015973:Gene Expression Regulation, Leukemic; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D000074008:MDS1 and EVI1 Complex Locus Protein; D008297:Male; D008875:Middle Aged; D015337:Multicenter Studies as Topic; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000097:blood / Q000235:genetics; D018365:Neoplasm, Residual; D011519:Proto-Oncogenes / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000097:blood; D012334:RNA, Neoplasm / Q000096:biosynthesis* / Q000097:blood; D016032:Randomized Controlled Trials as Topic; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014157:Transcription Factors / Q000096:biosynthesis* / Q000097:blood / Q000235:genetics; D014158:Transcription, Genetic; D055815:Young Adult
chemical_list: D004268:DNA-Binding Proteins; D000074008:MDS1 and EVI1 Complex Locus Protein; C063575:MECOM protein, human; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D014157:Transcription Factors
keywords: Acute myeloid leukemia; EVI1 overexpression; MLL; RT-qPCR