Molecular characterization of Waldenstrom's macroglobulinemia reveals frequent occurrence of two B-cell clones having distinct IgH VDJ sequences.
OBJECTIVE
Malignant B lineage cells in Waldenstrom's macroglobulinemia (WM) express a unique clonotypic IgM VDJ. The occurrence of biclonal B cells and their clonal relationships were characterized.


METHODS
Bone marrow and blood from 20 WM patients were analyzed for clonotypic VDJ sequences, clonal B-cell frequencies, and the complementary determining region 3 profile.


RESULTS
Two different clonotypic VDJ sequences were identified in 4 of 20 WM. In two cases, partner clones had different VDJ rearrangements, with one clonotypic signature in bone marrow and a second in blood. For both cases, the bone marrow clone was hypermutated, whereas the blood clone was germ line or minimally mutated. In two other cases, partner clones shared a common VDJ rearrangement but had different patterns of somatic mutations. They lacked intraclonal diversity and were more abundant in bone marrow than in blood. VDJ mutation profiles suggested they arose from a common IgM progenitor. Single-cell analysis in one case indicated the partner clones were reciprocally expressed, following rules of allelic exclusion.


CONCLUSIONS
The existence of two B-cell clones having distinct VDJ sequences is common in WM, suggesting that frequent transformation events may occur. In two cases, the partner clones had distinct tissue distributions in either blood or bone marrow, were of different immunoglobulin isotypes, and in one case exhibited differential response to therapy. The contributions of each clone are unknown. Their presence suggests that WM may involve a background of molecular and cellular events leading to emergence of one or more malignant clones.
Kriangkum
Jitra
J
Departments of Oncology, Cross Cancer Institute and Medicine, University of Alberta, Edmonton, Canada.
Taylor
Brian J
BJ
Treon
Steven P
SP
Mant
Michael J
MJ
Reiman
Tony
T
Belch
Andrew R
AR
Pilarski
Linda M
LM
D000368:Aged; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology*; D001483:Base Sequence; D019070:Cell Lineage; D002469:Cell Separation; D002999:Clone Cells; D022801:Complementarity Determining Regions / Q000235:genetics*; D019075:Electrophoresis, Capillary; D005260:Female; D005434:Flow Cytometry; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain; D050438:Genes, Immunoglobulin Heavy Chain; D006412:Hematopoietic Stem Cells; D006801:Humans; D007075:Immunoglobulin M / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D008258:Waldenstrom Macroglobulinemia / Q000235:genetics*
D022801:Complementarity Determining Regions; D007075:Immunoglobulin M