Identification of translocation products but not K-RAS mutations in memory B cells from patients with multiple myeloma.
BACKGROUND
Several laboratories have shown that cells with a memory B-cell phenotype can have the same clonotype as multiple myeloma tumor cells.


METHODS
The aim of this study was to determine whether some memory B cells have the same genetic alterations as their corresponding multiple myeloma malignant plasma cells. The methodology included sorting multiple myeloma or memory B cells into RNA stabilizing medium for generation of subset-specific polymerase chain reaction complementary DNA libraries from one or 100 cells.


RESULTS
Cells with the phenotype of tumor plasma cells (CD38(++)CD19(-)CD45(-/+)CD56(-/+/++)) or memory B cells (CD38(-)/CD19(+)/CD27(+)) were isolated by flow activated cell sorting. In samples from all four patients with multiple myeloma and from two of the three with monoclonal gammopathy of undetermined significance, we identified memory B cells expressing multiple myeloma-specific oncogenes (FGFR3; IGH-MMSET; CCND1 high) dysregulated by an IGH translocation in the respective tumor plasma cells. By contrast, in seven patients with multiple myeloma, each of whom had tumor plasma cells with a K-RAS61 mutation, a total of 32,400 memory B cells were analyzed using a sensitive allele-specific, competitive blocker polymerase chain reaction assay, but no K-RAS mutations were identified.


CONCLUSIONS
The increased expression of a specific "early" oncogene of multiple myeloma (monoclonal gammopathy of undetermined significance) in some memory B cells suggests that dysregulation of the oncogene occurs in a precursor B-cell that can generate memory B cells and transformed plasma cells. However, if memory B cells lack "late" oncogene (K-RAS) mutations but express the "early" oncogene, they cannot be involved in maintaining the multiple myeloma tumor, but presumably represent a clonotypic remnant that is only partially transformed.
1Department of Hematology, Herlev Hospital, University of Copenhagen, Herlev, Denmark.
Thomas
T
Rasmussen
Jacob
J
Haaber
Inger Marie
IM
Dahl
Lene M
LM
Knudsen
Gitte B
GB
Kerndrup
Marianne
M
Lodahl
Hans E
HE
Johnsen
Michael
M
Kuehl
D001402:B-Lymphocytes / Q000473:pathology*; D002999:Clone Cells / Q000473:pathology; D011905:Genes, ras / Q000235:genetics*; D006801:Humans; D007156:Immunologic Memory; D009101:Multiple Myeloma / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D009154:Mutation; D014178:Translocation, Genetic