1. A Next-Generation Sequencing Strategy for Evaluating the Most Common Genetic Abnormalities in Multiple Myeloma
- Author
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Noemi Puig, Marcos González, Albert Oriol, Miguel Alcoceba, Ana Isabel Teruel, Gonzalo R. Ordóñez, Luis A. Corchete, David Castillo, Joaquin Martinez-Lopez, Joan Bladé, Jesús F. San Miguel, María Jara-Acevedo, Laura Rosiñol, Ana Balanzategui, Juan J. Lahuerta, Alberto Orfao, María García-Álvarez, Norma C. Gutiérrez, María C. Chillón, Luis Palomera, Cristina Jimenez, María Eugenia Sarasquete, Ramón García-Sanz, Maria V. Mateos, and María Isabel Prieto-Conde
- Subjects
Male ,0301 basic medicine ,Neuroblastoma RAS viral oncogene homolog ,DNA Mutational Analysis ,Single-nucleotide polymorphism ,Biology ,medicine.disease_cause ,Pathology and Forensic Medicine ,03 medical and health sciences ,0302 clinical medicine ,Gene Frequency ,hemic and lymphatic diseases ,medicine ,Humans ,HRAS ,Multiple myeloma ,Aged ,Genetics ,medicine.diagnostic_test ,Breakpoint ,High-Throughput Nucleotide Sequencing ,Middle Aged ,medicine.disease ,Minimal residual disease ,030104 developmental biology ,Molecular Diagnostic Techniques ,030220 oncology & carcinogenesis ,Mutation ,Molecular Medicine ,Female ,KRAS ,Multiple Myeloma ,Genes, Neoplasm ,Fluorescence in situ hybridization - 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.
- Published
- 2017