Clinical Lymphoma, Myeloma and Leukemia Characteristics of Waldenström Macroglobulinemia in Koreans According to Mutational Status of MYD88 and CXCR4: Analysis by Ultra-Deep Sequencing  Dong Woo Shin, Sung-Min Kim, Jung-Ah Kim, Hee Sue Park, Sang Mee Hwang, Kyongok Im, Sungsik Kim, Jinhyun Kim, Sunghoon Kwon, Sung-Soo Yoon, Dong Soon Lee  Clinical Lymphoma, Myeloma and Leukemia  DOI: 10.1016/j.clml.2019.03.009 Copyright © 2019 Terms and Conditions

Figure 1 CXCR4 mutations detected by ultra-deep next-generation sequencing. Each of the six mutations identified were positioned in the C-terminal domain of CXCR4, but none in the N-terminal domain and transmembrane domain. Two variants were novel (a marked), and all of Waldenström Macroglobulinemia patients with CXCR4 mutation also harbored the MYD88L265P mutation. None of the six mutations were detected by Sanger sequencing. Variant allele frequencies of each case were 23.8%, 7.9%, 12.6%, 3.2%, 7.6%, and 41.7%, respectively. Read depths of each case were 2,099, 6,270, 2,868, 5,029, 21,139, and 4,456, respectively. Clinical Lymphoma, Myeloma and Leukemia DOI: (10.1016/j.clml.2019.03.009) Copyright © 2019 Terms and Conditions

Figure 2 Illustration of molecular and cytogenetic abnormalities in patients with Waldenström macroglobulinemia. Among 37 patients, neither molecular nor cytogenetic analysis was available in 3 patients due to lack of specimens. Each horizontal number means each patient (No. 1 to 34) and each vertical line indicates the status of mutations and cytogenetic aberrations. The gray square indicates an absence of changes for each category, and the asterisk (*) indicates that the results were not available. Clinical Lymphoma, Myeloma and Leukemia DOI: (10.1016/j.clml.2019.03.009) Copyright © 2019 Terms and Conditions

Figure 3 Flow chart to detect MYD88 mutation across the different cell lineages by single cell genomic study. Bone marrow aspirates slide was scanned by automated microscope. Using a custom software, white blood cells in the scanned image were classified into plasma cells, plasmacytoid lymphocytes, lymphocytes and neutrophils, and their locations were marked by the hematologist. Isolated single cells were lysed and amplified by PCR and the sequences were analyzed by capillary sequencing. In all of 3 patients, MYD88L265P was detected on plasma cells, plasmacytoid lymphocytes, and lymphocytes but it was not detected on neutrophils. Clinical Lymphoma, Myeloma and Leukemia DOI: (10.1016/j.clml.2019.03.009) Copyright © 2019 Terms and Conditions

Figure 4 Kaplan-Meier survival curve of 31 WM patients classified by MYD88 and CXCR4 mutation status. Group 1 showed adverse survival and 1 year survival rate of group 1 (66.7%) was lower than group 2 (94.7%), though it was not statistically significant (P=0.410). 3 years and 8 years survival rate of group 2 was 81.2% and 40.6%, respectively. There were no death events in group 3 patients during the research period. Clinical Lymphoma, Myeloma and Leukemia DOI: (10.1016/j.clml.2019.03.009) Copyright © 2019 Terms and Conditions