Jiménez Cristina, Jara-Acevedo María, Corchete Luis A, Castillo David, Ordóñez Gonzalo R, Sarasquete María E, Puig Noemí, Martínez-López Joaquín, Prieto-Conde María I, García-Álvarez María, Chillón María C, Balanzategui Ana, Alcoceba Miguel, Oriol Albert, Rosiñol Laura, Palomera Luis, Teruel Ana I, Lahuerta Juan J, Bladé Joan, Mateos María V, Orfão Alberto, San Miguel Jesús F, González Marcos, Gutiérrez Norma C, García-Sanz Ramón
Hematology Department, University Hospital of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
DNA Sequencing Service, University of Salamanca, Research Biomedical Institute of Salamanca (IBSAL), Salamanca, Spain.
J Mol Diagn. 2017 Jan;19(1):99-106. doi: 10.1016/j.jmoldx.2016.08.004. Epub 2016 Nov 15.
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.
基因改变的识别与特征分析对于多发性骨髓瘤的诊断至关重要，并且可能指导治疗决策。目前，骨髓瘤的基因组分析要涵盖具有预后影响的多种改变，需要荧光原位杂交（FISH）、单核苷酸多态性阵列和测序技术，这些技术成本高昂、 labor intensive且需要大量浆细胞。为克服这些局限性，我们设计了一种靶向捕获二代测序方法，用于一步鉴定IGH易位、V(D)J克隆重排、IgH同种型和体细胞突变，以快速识别风险组和特定的可靶向分子病变。用该检测板对48例新诊断的骨髓瘤患者进行了检测，该检测板包括IGH以及在骨髓瘤中经常发生突变的6个基因：NRAS、KRAS、HRAS、TP53、MYC和BRAF。我们鉴定出了先前通过FISH检测到的17种IGH易位中的14种，以及3种FISH未检测到的已确认易位，还有断点识别的额外优势，可将其用作评估微小残留病的靶点。分别在77%和65%的患者中鉴定出了IgH亚类和V(D)J重排。突变分析显示，48例患者中有16例（33%）在至少一个评估基因中存在错义蛋白编码改变。该方法可能是一种用于多发性骨髓瘤分子特征分析的具有时间和成本效益的诊断方法。
