PMID- 31605801 OWN - NLM STAT- MEDLINE DCOM- 20210510 LR - 20210510 IS - 1943-7811 (Electronic) IS - 1525-1578 (Linking) VI - 22 IP - 1 DP - 2020 Jan TI - A New Next-Generation Sequencing Strategy for the Simultaneous Analysis of Mutations and Chromosomal Rearrangements at DNA Level in Acute Myeloid Leukemia Patients. PG - 60-71 LID - S1525-1578(19)30393-9 [pii] LID - 10.1016/j.jmoldx.2019.08.002 [doi] AB - Acute myeloid leukemias (AMLs) are currently genomically characterized by karyotype, fluorescence in situ hybridization (FISH), real-time quantitative PCR, and DNA sequencing. Next-generation sequencing offers the promise of detecting all genomic lesions in a single run. However, technical limitations have hampered the detection of chromosomal rearrangements, so most studies are limited to somatic mutation assessment or require the use of RNA-based strategies. To overcome these limitations, we designed a targeted-DNA capture next-generation sequencing approach associated with easy-to-perform public bioinformatic tools for one-step identification of translocations, inversions, and somatic mutations in AML. Thirty well-characterized newly diagnosed myeloid leukemia patients (27 AML and 3 chronic myeloid leukemia) were tested with the panel. Twenty-three of 24 known rearrangements, as well as one novel fusion gene that could not be detected by karyotype/fluorescence in situ hybridization/real-time quantitative PCR, were detected. This strategy also identified all chromosomal breakpoints as potential targets for future high-sensitive minimal residual disease studies. In addition, mutation analysis revealed the presence of missense protein-coding alterations in at least 1 of the 32 genes evaluated in 21 of 30 patients (70%). This strategy may represent a time- and cost-effective diagnostic method for molecular characterization in AML. CI - Copyright (c) 2020 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved. FAU - Prieto-Conde, M Isabel AU - Prieto-Conde MI AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Corchete, Luis A AU - Corchete LA AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Garcia-Alvarez, Maria AU - Garcia-Alvarez M AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Jimenez, Cristina AU - Jimenez C AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Medina, Alejandro AU - Medina A AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Balanzategui, Ana AU - Balanzategui A AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Hernandez-Ruano, Montserrat AU - Hernandez-Ruano M AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Maldonado, Rebeca AU - Maldonado R AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Sarasquete, M Eugenia AU - Sarasquete ME AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Alcoceba, Miguel AU - Alcoceba M AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Puig, Noemi AU - Puig N AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Gonzalez-Calle, Veronica AU - Gonzalez-Calle V AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Garcia-Sanz, Ramon AU - Garcia-Sanz R AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. Electronic address: rgarcias@usal.es. FAU - Gutierrez, Norma C AU - Gutierrez NC AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Gonzalez-Diaz, Marcos AU - Gonzalez-Diaz M AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. FAU - Chillon, M Carmen AU - Chillon MC AD - Department of Hematology, University Hospital of Salamanca (HUS/IBSAL), CIBERONC and Center for Cancer Research-IBMCC (USAL-CSIC), Salamanca, Spain. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PT - Validation Study DEP - 20191009 PL - United States TA - J Mol Diagn JT - The Journal of molecular diagnostics : JMD JID - 100893612 RN - 9007-49-2 (DNA) SB - IM MH - Base Sequence MH - Bone Marrow MH - *Chromosome Aberrations MH - Chromosome Breakpoints MH - DNA/*genetics MH - DNA Mutational Analysis/methods MH - Data Accuracy MH - High-Throughput Nucleotide Sequencing/*methods MH - Humans MH - In Situ Hybridization, Fluorescence/methods MH - Karyotyping/methods MH - Leukemia, Myelogenous, Chronic, BCR-ABL Positive/*genetics MH - Leukemia, Myeloid, Acute/*genetics MH - *Mutation, Missense MH - Real-Time Polymerase Chain Reaction/methods MH - Sensitivity and Specificity MH - Sequence Analysis, DNA/methods EDAT- 2019/10/13 06:00 MHDA- 2021/05/11 06:00 CRDT- 2019/10/13 06:00 PHST- 2019/04/02 00:00 [received] PHST- 2019/07/23 00:00 [revised] PHST- 2019/08/14 00:00 [accepted] PHST- 2019/10/13 06:00 [pubmed] PHST- 2021/05/11 06:00 [medline] PHST- 2019/10/13 06:00 [entrez] AID - S1525-1578(19)30393-9 [pii] AID - 10.1016/j.jmoldx.2019.08.002 [doi] PST - ppublish SO - J Mol Diagn. 2020 Jan;22(1):60-71. doi: 10.1016/j.jmoldx.2019.08.002. Epub 2019 Oct 9.