PMID- 25807917 OWN - NLM STAT- MEDLINE DCOM- 20150929 LR - 20210224 IS - 1934-6638 (Electronic) IS - 1934-662X (Linking) VI - 123 IP - 7 DP - 2015 Jul TI - Multiplex sequencing for EZH2, CD79B, and MYD88 mutations using archival cytospin preparations from B-cell non-Hodgkin lymphoma aspirates previously tested for MYC rearrangement and IGH/BCL2 translocation. PG - 413-20 LID - 10.1002/cncy.21535 [doi] AB - BACKGROUND: Gene rearrangements and specific translocations define some B-cell non-Hodgkin lymphoma (NHL) subtypes. Genome-wide mutational studies have revealed recurrent point mutations with prognostic implications. The goals of this study were to evaluate the feasibility of applying a multiplex mutation assay to archival cytospin preparations (CPs) and to investigate the rate of EZH2, CD79B, and MYD88 mutations in B-cell NHL samples previously tested for MYC rearrangement and/or IGH/BCL-2 translocation. METHODS: DNA was extracted from archival CPs of B-cell NHL cases with previous fluorescence in situ hybridization (FISH) assays for MYC rearrangement and/or IGH/BCL-2 translocation. Multiplex sequencing was performed for the detection of EZH2 (Y641), CD79B (Y196), and MYD88 (L265) mutations. Sanger sequencing was applied to samples with positive results and failed assays. RESULTS: Eighty-eight archival CPs were available from 40 patients. Alterations detected by FISH were: MYC rearrangement (10 cases), IGH/BCL-2 translocations (21 cases), dual translocations (6 cases), and other abnormalities for IGH/BCL-2 (23 cases) and for MYC (16 cases). DNA concentration ranged from 1.88 to 62.85 ng/microL (mean, 9.46 ng/microL). Successful results were obtained in 88.0% of the specimens submitted to multiplex sequencing. With Sanger sequencing, 2 additional mutated cases were found, and all cases with mutations were confirmed. Eight specimens showed mutations: 6 for EZH2, 1 for CD79B, and 1 for MYD88. Among them, 5 cases showed concurrent MYC and/or IGH/BCL-2 translocations and 2 revealed abnormal signals of IGH/BCL-2 and MYC. CONCLUSIONS: CPs archived for up to 6 years are a reliable source of high-quality genomic material for multiplex sequencing. Almost all B-cell NHL with point mutations showed concurrent chromosomal abnormalities. CI - (c) 2015 American Cancer Society. FAU - Santos, Gilda da Cunha AU - Santos Gda C AD - Department of Laboratory Medicine and Pathobiology, University of Toronto, Ontario, Canada. AD - Laboratory Medicine Program, University Health Network, Toronto, Ontario, Canada. FAU - Saieg, Mauro Ajaj AU - Saieg MA AD - Department of Pathology, Santa Casa Medical School, Sao Paulo, Brazil. AD - Fleury Laboratories, Sao Paulo, Brazil. FAU - Ko, Hyang Mi AU - Ko HM AD - Department of Laboratory Medicine and Pathobiology, University of Toronto, Ontario, Canada. AD - Laboratory Medicine Program, University Health Network, Toronto, Ontario, Canada. FAU - Geddie, William R AU - Geddie WR AD - Department of Laboratory Medicine and Pathobiology, University of Toronto, Ontario, Canada. AD - Laboratory Medicine Program, University Health Network, Toronto, Ontario, Canada. FAU - Boerner, Scott L AU - Boerner SL AD - Department of Laboratory Medicine and Pathobiology, University of Toronto, Ontario, Canada. AD - Laboratory Medicine Program, University Health Network, Toronto, Ontario, Canada. FAU - Craddock, Kenneth J AU - Craddock KJ AD - Laboratory Medicine Program, University Health Network, Toronto, Ontario, Canada. FAU - Crump, Michael AU - Crump M AD - Division of Medical Oncology and Hematology, University Health Network, Toronto, Ontario, Canada. FAU - Bailey, Denis AU - Bailey D AD - Department of Laboratory Medicine and Pathobiology, University of Toronto, Ontario, Canada. AD - Laboratory Medicine Program, University Health Network, Toronto, Ontario, Canada. LA - eng GR - Canadian Institutes of Health Research/Canada PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150323 PL - United States TA - Cancer Cytopathol JT - Cancer cytopathology JID - 101499453 RN - 0 (CD79 Antigens) RN - 0 (CD79B protein, human) RN - 0 (MYD88 protein, human) RN - 0 (Myeloid Differentiation Factor 88) RN - 0 (Phosphoproteins) RN - 0 (Proto-Oncogene Proteins c-bcl-2) RN - 0 (Proto-Oncogene Proteins c-myc) RN - 0 (SPECC1L protein, human) RN - EC 2.1.1.43 (EZH2 protein, human) RN - EC 2.1.1.43 (Enhancer of Zeste Homolog 2 Protein) RN - EC 2.1.1.43 (Polycomb Repressive Complex 2) SB - IM MH - Adult MH - Aged MH - Biopsy, Fine-Needle MH - Biopsy, Needle MH - CD79 Antigens/*chemistry/genetics MH - Cohort Studies MH - Databases, Factual MH - Enhancer of Zeste Homolog 2 Protein MH - Female MH - Gene Rearrangement MH - Humans MH - In Situ Hybridization, Fluorescence MH - Lymphoma, B-Cell/*genetics/pathology MH - Male MH - Mass Spectrometry/methods MH - Middle Aged MH - Myeloid Differentiation Factor 88/*chemistry/genetics MH - Phosphoproteins/chemistry MH - Point Mutation/*genetics MH - Polycomb Repressive Complex 2/*chemistry/genetics MH - Predictive Value of Tests MH - Proto-Oncogene Proteins c-bcl-2/chemistry MH - Proto-Oncogene Proteins c-myc/genetics MH - Retrospective Studies MH - Sensitivity and Specificity MH - Sequence Analysis, DNA/*methods MH - Specimen Handling/*methods MH - Translocation, Genetic OTO - NOTNLM OT - B-cell non-Hodgkin lymphoma OT - CD79B OT - DNA mutational analysis OT - EZH2 OT - IGH/BCL2 OT - MYC OT - MYD88 OT - MassARRAY spectrometry OT - cytospin preparation OT - fine needle biopsy EDAT- 2015/03/27 06:00 MHDA- 2015/09/30 06:00 CRDT- 2015/03/27 06:00 PHST- 2014/12/10 00:00 [received] PHST- 2015/01/29 00:00 [revised] PHST- 2015/02/10 00:00 [accepted] PHST- 2015/03/27 06:00 [entrez] PHST- 2015/03/27 06:00 [pubmed] PHST- 2015/09/30 06:00 [medline] AID - 10.1002/cncy.21535 [doi] PST - ppublish SO - Cancer Cytopathol. 2015 Jul;123(7):413-20. doi: 10.1002/cncy.21535. Epub 2015 Mar 23.