title: CEBPA-double-mutated acute myeloid leukemia displays a unique phenotypic profile: a reliable screening method and insight into biological features.
abstract: Mutations in CCAAT/enhancer binding protein α (CEBPA) occur in 5-10% of cases of acute myeloid leukemia.CEBPA-double-mutated cases usually bear biallelic N- and C-terminal mutations and are associated with a favorable clinical outcome. Identification ofCEBPAmutants is challenging because of the variety of mutations, intrinsic characteristics of the gene and technical issues. Several screening methods (fragment-length analysis, gene expression array) have been proposed especially for large-scale clinical use; although efficient, they are limited by specific concerns. We investigated the phenotypic profile of blast and maturing bone marrow cell compartments at diagnosis in 251 cases of acute myeloid leukemia. In this cohort, 16 (6.4%) patients had twoCEBPAmutations, whereas ten (4.0%) had a single mutation. First, we highlighted that theCEBPA-double-mutated subset displays recurrent phenotypic abnormalities in all cell compartments. By mutational analysis after cell sorting, we demonstrated that this common phenotypic signature depends onCEBPA-double-mutated multi-lineage involvement. From a multidimensional study of phenotypic data, we developed a classifier including ten core and widely available parameters. The selected markers on blasts (CD34, CD117, CD7, CD15, CD65), neutrophil (SSC, CD64), monocytic (CD14, CD64) and erythroid (CD117) compartments were able to clusterCEBPA-double-mutated cases. In a validation set of 259 AML cases from three independent centers, our classifier showed excellent performance with 100% specificity and 100% sensitivity. We have, therefore, established a reliable screening method, based upon multidimensional analysis of widely available phenotypic parameters. This method provides early results and is suitable for large-scale detection ofCEBPA-double-mutated status, allowing gene sequencing to be focused in selected cases.
authors.affiliation: Unità Funzionale di Ematologia, Università degli Studi, AOU Careggi, Firenze, Italy francesco.mannelli@unifi.it.|Istituto Toscano Tumori, Firenze, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Mannelli
authors.affiliation: Unità Funzionale di Ematologia, Università degli Studi, AOU Careggi, Firenze, Italy.|Istituto Toscano Tumori, Firenze, Italy.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Ponziani
authors.affiliation: Unità Funzionale di Ematologia, Università degli Studi, AOU Careggi, Firenze, Italy.|Istituto Toscano Tumori, Firenze, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Bencini
authors.affiliation: Unità Funzionale di Ematologia, Università degli Studi, AOU Careggi, Firenze, Italy.|Istituto Toscano Tumori, Firenze, Italy.
authors.forename: Maria Ida
authors.initials: MI
authors.lastname: Bonetti
authors.affiliation: SOD Diagnostica Genetica, AOU Careggi, Firenze, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Benelli
authors.affiliation: Unità Funzionale di Ematologia, Università degli Studi, AOU Careggi, Firenze, Italy.|Istituto Toscano Tumori, Firenze, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Cutini
authors.affiliation: Unità Funzionale di Ematologia, Università degli Studi, AOU Careggi, Firenze, Italy.|Istituto Toscano Tumori, Firenze, Italy.
authors.forename: Giacomo
authors.initials: G
authors.lastname: Gianfaldoni
authors.affiliation: Unità Funzionale di Ematologia, Università degli Studi, AOU Careggi, Firenze, Italy.|Istituto Toscano Tumori, Firenze, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Scappini
authors.affiliation: Unità Funzionale di Ematologia, Università degli Studi, AOU Careggi, Firenze, Italy.|Istituto Toscano Tumori, Firenze, Italy.
authors.forename: Fabiana
authors.initials: F
authors.lastname: Pancani
authors.affiliation: Unità Funzionale di Ematologia, Università degli Studi, AOU Careggi, Firenze, Italy.|Istituto Toscano Tumori, Firenze, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Piccini
authors.affiliation: SOD Laboratorio Centrale, Settore Citometria Clinica, AOU Careggi, Firenze, Italy.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Rondelli
authors.affiliation: SOD Laboratorio Centrale, Settore Citometria Clinica, AOU Careggi, Firenze, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Caporale
authors.affiliation: SOD Laboratorio Centrale, Settore Citometria Clinica, AOU Careggi, Firenze, Italy.
authors.forename: Anna Maria Grazia
authors.initials: AM
authors.lastname: Gelli
authors.affiliation: SOD Laboratorio Centrale, Settore Citometria Clinica, AOU Careggi, Firenze, Italy.
authors.forename: Benedetta
authors.initials: B
authors.lastname: Peruzzi
authors.affiliation: Centro di Ricerca Emato-Oncologica AIL (CREA), Spedali Civili, Brescia, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Chiarini
authors.affiliation: Divisione di Ematologia, Spedali Civili, Brescia, Italy.
authors.forename: Erika
authors.initials: E
authors.lastname: Borlenghi
authors.affiliation: Unità Strutturale Complessa di Ematologia, Ospedali Riuniti, Bergamo, Italy.
authors.forename: Orietta
authors.initials: O
authors.lastname: Spinelli
authors.affiliation: Unità Strutturale Complessa di Ematologia, Ospedali Riuniti, Bergamo, Italy.
authors.forename: Damiano
authors.initials: D
authors.lastname: Giupponi
authors.affiliation: Unità Strutturale Complessa di Ematologia, Ospedali Riuniti, Bergamo, Italy.
authors.forename: Pamela
authors.initials: P
authors.lastname: Zanghì
authors.affiliation: Divisione di Ematologia, Ospedale dell'Angelo & Ospedale SS. Giovanni e Paolo, Mestre-Venezia, Italy.
authors.forename: Renato
authors.initials: R
authors.lastname: Bassan
authors.affiliation: Unità Strutturale Complessa di Ematologia, Ospedali Riuniti, Bergamo, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Rambaldi
authors.affiliation: Divisione di Ematologia, Spedali Civili, Brescia, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Rossi
authors.affiliation: Unità Funzionale di Ematologia, Università degli Studi, AOU Careggi, Firenze, Italy.|Istituto Toscano Tumori, Firenze, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Bosi
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor; D001853:Bone Marrow / Q000473:pathology; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics* / Q000378:metabolism; D016000:Cluster Analysis; D020732:Cytogenetic Analysis; D004252:DNA Mutational Analysis; D005260:Female; D056726:Genetic Association Studies; D005838:Genotype; D006801:Humans; D016130:Immunophenotyping; D053208:Kaplan-Meier Estimate; D015470:Leukemia, Myeloid, Acute / Q000175:diagnosis* / Q000235:genetics* / Q000401:mortality; D008297:Male; D008875:Middle Aged; D009154:Mutation; D010641:Phenotype; D011379:Prognosis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D014408:Biomarkers, Tumor; D022763:CCAAT-Enhancer-Binding Protein-alpha