title: A Next-Generation Sequencing Strategy for Evaluating the Most Common Genetic Abnormalities in Multiple Myeloma.
abstract: Identification and characterization of genetic alterations are essential for diagnosis of multiple myeloma and may guide therapeutic decisions. Currently, genomic analysis of myeloma to cover the diverse range of alterations with prognostic impact requires fluorescence in situ hybridization (FISH), single nucleotide polymorphism arrays, and sequencing techniques, which are costly and labor intensive and require large numbers of plasma cells. To overcome these limitations, we designed a targeted-capture next-generation sequencing approach for one-step identification of IGH translocations, V(D)J clonal rearrangements, the IgH isotype, and somatic mutations to rapidly identify risk groups and specific targetable molecular lesions. Forty-eight newly diagnosed myeloma patients were tested with the panel, which included IGH and six genes that are recurrently mutated in myeloma: NRAS, KRAS, HRAS, TP53, MYC, and BRAF. We identified 14 of 17 IGH translocations previously detected by FISH and three confirmed translocations not detected by FISH, with the additional advantage of breakpoint identification, which can be used as a target for evaluating minimal residual disease. IgH subclass and V(D)J rearrangements were identified in 77% and 65% of patients, respectively. Mutation analysis revealed the presence of missense protein-coding alterations in at least one of the evaluating genes in 16 of 48 patients (33%). This method may represent a time- and cost-effective diagnostic method for the molecular characterization of multiple myeloma.
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Jiménez
authors.affiliation: DNA Sequencing Service, University of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Jara-Acevedo
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: Luis A
authors.initials: LA
authors.lastname: Corchete
authors.affiliation: DREAMgenics, Oviedo, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Castillo
authors.affiliation: DREAMgenics, Oviedo, Spain.
authors.forename: Gonzalo R
authors.initials: GR
authors.lastname: Ordóñez
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: María E
authors.initials: ME
authors.lastname: Sarasquete
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: Noemí
authors.initials: N
authors.lastname: Puig
authors.affiliation: Hematology Department, 12 de Octubre Hospital, Unit of Cancer Research Innovation Spain (CRIS), Spanish National Cancer Research Center (CNIO), University of Madrid, Madrid, Spain.
authors.forename: Joaquín
authors.initials: J
authors.lastname: Martínez-López
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: María I
authors.initials: MI
authors.lastname: Prieto-Conde
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: María
authors.initials: M
authors.lastname: García-Álvarez
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: María C
authors.initials: MC
authors.lastname: Chillón
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Balanzategui
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Alcoceba
authors.affiliation: Catalan Institute of Oncology, Josep Carreras Institute, Germans Trias i Pujol Hospital, Barcelona, Spain.
authors.forename: Albert
authors.initials: A
authors.lastname: Oriol
authors.affiliation: Research Biomedical Institute August Pi i Sunyer, Clinical Hospital of Barcelona, Barcelona, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Rosiñol
authors.affiliation: Lozano Blesa Hospital, Zaragoza, Spain.
authors.forename: Luis
authors.initials: L
authors.lastname: Palomera
authors.affiliation: Clincal Hospital of Valencia, Valencia, Spain.
authors.forename: Ana I
authors.initials: AI
authors.lastname: Teruel
authors.affiliation: Hematology Department, 12 de Octubre Hospital, Unit of Cancer Research Innovation Spain (CRIS), Spanish National Cancer Research Center (CNIO), University of Madrid, Madrid, Spain.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Lahuerta
authors.affiliation: Research Biomedical Institute August Pi i Sunyer, Clinical Hospital of Barcelona, Barcelona, Spain.
authors.forename: Joan
authors.initials: J
authors.lastname: Bladé
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: María V
authors.initials: MV
authors.lastname: Mateos
authors.affiliation: DNA Sequencing Service, University of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: Alberto
authors.initials: A
authors.lastname: Orfão
authors.affiliation: Center for Applied Medical Research, University of Navarra Hospital, Institute of Health Research of Navarra (IDISNA), Pamplona, Spain.
authors.forename: Jesús F
authors.initials: JF
authors.lastname: San Miguel
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain. Electronic address: rgarcias@usal.es.
authors.forename: Marcos
authors.initials: M
authors.lastname: González
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: Norma C
authors.initials: NC
authors.lastname: Gutiérrez
authors.affiliation: Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
authors.forename: Ramón
authors.initials: R
authors.lastname: García-Sanz
mesh_terms: D000368:Aged; D004252:DNA Mutational Analysis / Q000379:methods*; D005260:Female; D005787:Gene Frequency; D052138:Genes, Neoplasm; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques; D009101:Multiple Myeloma / Q000175:diagnosis / Q000235:genetics*; D009154:Mutation