|
|||||||||
|
|
|
|
|
|
|
|
|
|
|
NEOPLASIA
From the Department of Hematology and Internal Medicine
and the Department of Laboratory Medicine and Pathology, Mayo Clinic,
Rochester, MN; and the National Cancer Institute, Genetics Branch,
Bethesda MD.
Lymphoplasmacytic lymphoma (LPL) is characterized by
t(9;14)(p13;q32) in 50% of patients who lack paraproteinemia.
Waldenström macroglobulinemia (WM), which has an immunoglobulin M
(IgM) paraproteinemia, is classified as an LPL. Rare reports have
suggested that WM sometimes is associated with 14q23 translocations,
deletions of 6q, and t(11;18)(q21;q21). We tested for these
abnormalities in the clonal cells of WM patients. We selected patients
with clinicopathologic diagnosis of WM (all had IgM levels greater than
1.5 g/dL). Southern blot assay was used to detect legitimate and
illegitimate IgH switch rearrangements. In addition to conventional
cytogenetic (CC) and multicolor metaphase fluorescence in situ
hybridization (M-FISH) analyses, we used interphase FISH to screen for
t(9;14)(p13;q32) and other IgH translocations, t(11;18)(q21;q21),
and 6q21 deletions. Genomic stability was also assessed using
chromosome enumeration probes for chromosomes 7, 9, 11, 12, 15, and 17 in 15 patients. There was no evidence of either legitimate or
illegitimate IgH rearrangements by Southern blot assay (n = 12). CC
(n = 37), M-FISH (n = 5), and interphase FISH (n = 42) failed to
identify IgH or t(11;18) translocations. Although tumor cells from most
patients were diploid for the chromosomes studied, deletions of 6q21
were observed in 42% of patients. In contrast to LPL tumors that are not associated with paraproteinemia and that have frequent
t(9;14)(p13;q32) translocations, IgH translocations are not found in
WM, a form of LPL tumor distinguished by IgM paraproteinemia. However,
WM tumor cells, which appear to be diploid or near diploid, often have
deletions of 6q21.
(Blood. 2002;100:2996-3001) Waldenström macroglobulinemia (WM) is a
B-cell lymphoproliferative disorder characterized by immunoglobulin M
(IgM) paraproteinemia and the accumulation of clonal lymphoplasmacytic
cells in the bone marrow (BM).1,2 With advancing disease,
patients may acquire organomegaly, anemia, and hyperviscosity. A serum
monoclonal IgM level that exceeds a concentration of 1.5 g/dL is
characteristic of WM and is useful in differentiating WM from other
neoplasms with plasmacytoid differentiation.3 The
diagnosis is based on the presence of the characteristic accompanying
clinical features and the pathologic findings, most commonly from the
bone marrow (Table 1). Because of its
morphologic and immunophenotypic features, the pathologic designation
for WM has been lymphoplasmacytic lymphoma (LPL), as proposed by the
revised European-American lymphoma classification.3 Because LPL is not always associated with paraproteinemia, it is
possible that LPL without paraproteinemia and WM (LPL with IgM
paraproteinemia) are distinct pathologic entities.
It has been reported that the t(9;14)(p13;q32) translocation occurs in
at least 50% of patients with LPL.4 The t(9;14)(p13;q32) results in the up-regulation of PAX-5,5,6
implicating this gene as a putative oncogene in the pathogenesis of
LPL.7-9 Other PAX genes are known to be
involved in diverse human neoplasia.10 PAX-5
encodes for the transcription factor, B-cell-specific activation protein (BSAP), which is a 50-kDa protein critical for B-cell development. BSAP up-regulation appears to cause an increase in B-cell
proliferation11 and is characteristically absent in normal and malignant plasma cells.12-15 BSAP/PAX-5 is
known to down-regulate the I The underlying genetic abnormalities responsible for WM have not been
identified. From a limited number of conventional cytogenetic (CC)
studies, no recurrent cytogenetic abnormalities have been detected in
tumor cells from patients with WM. However, there are sporadic reports
of WM tumors with IgH or t(11;18)(q21;q21) translocations or 6q
abnormalities, each of which occurs in other kinds of B-cell tumors.
Thus, we decided to test a cohort of patients with well-defined WM to
address 3 issues. First, despite the expression of IgM in WM, does
legitimate or illegitimate IgH switch recombination occur, and perhaps
mediate IgH translocations, as previously shown in multiple myeloma and
other B-cell tumors?20 Second, using a highly sensitive
and specific interphase fluorescence in situ hybridization (FISH)
assay, what is the incidence of the t(9;14)(p13;q32) or other IgH
translocations in WM? Third, through a combination of CC, multicolor
metaphase-FISH (M-FISH), and interphase FISH assays, what is the
incidence of recurrent numeric or structural karyotype
abnormalities Patients and bone marrow samples
In 19 patients, bone marrow research aspirates were further enriched by simultaneous CD138+ and CD19+ magnetic bead selection (Miltenyi Biotec, Auburn, CA). Owen et al21,22 have found that clonal cells in WM express CD19, CD138, or both. Because of the pleomorphic nature of the clonal process in WM, we thought that with the combination of CD138 and the B-cell marker CD19, we could enrich for the clonal cells of patients. When we only had stored BM samples previously cultured for metaphase analysis, it was not possible to perform magnetic bead selection or concurrent immunofluorescence staining of the cytoplasmic immunoglobulins for those patients. This study was conducted under the approval of the Mayo Clinic institutional review board, and patients gave informed consent for the sample collection. The study was conducted in accordance with the Declaration of Helsinki for research with human subjects. Southern blot analysis Southern blot analysis was performed according to our previously published technique.20 In brief, we used a JH probe to detect the germ line and clonal bands to ensure the presence of clonal DNA. We subsequently performed sequential hybridization with probes flanking the 5' and the 3' ends of the Sµ switch region. The presence of fully concordant 5'Sµ and 3'Sµ bands indicates the absence of IgH switch recombination. Alternatively, discordant 5'Sµ and 3'Sµ bands that do not hybridize with other IgH switch probes strongly suggest illegitimate IgH switch recombination caused by a chromosomal translocation.20 A total of 2.5 to 10 µg BM DNA (these samples were not magnetic microbead-enriched) was loaded into each well after complete digestion with restriction enzyme HindIII (Figure 1)
Standard cytogenetic analysis Conventional karyotypes performed for clinical evaluation were available for 37 patients. BM specimens were processed by direct and short-term culture techniques, as described previously.23 At least 20 banded metaphases were analyzed for each patient, with representative karyotypes prepared from at least 2 metaphases from each clone. The karyotype was described according to the International System for Human Cytogenetics Nomenclature (ISCN, 1995).Multicolor-FISH assay M-FISH was performed as previously described.24,25 Slides with informative karyotypes in 5 patients were processed using the Spectra Vysion (Vysis, Downer's Grove, IL) reagent. Probes for M-FISH were placed on the hybridization site, protected with a coverslip, sealed with rubber cement, and placed in the HYBrite (Vysis) for codenaturation. Slides were viewed with a Zeiss (Thornwood, NY) microscope powered by a 100-W mercury bulb. Filter sets for capturing M-FISH images and viewing metaphases were from Vysis.Interphase FISH We used 2 different probe strategies to detect IgH rearrangements by interphase FISH. We first used a break-apart strategy (segregation), using a cosmid probe (VH) (labeled red) specific for the IgH variable region and a BAC clone (CH) specific for the IgH constant (labeled green).26 Under separate experiments and to detect the t(9;14)(p13;q32) translocation, we used a fusion strategy (colocalization), using a BAC clone (clone 112gO2; Incyte Genomics, Palo Alto, CA) specific for the PAX-5 gene at 9p13 (labeled red) (Figure 2), together with the VH/CH probes (both labeled in green).26 To validate these probes, we tested them in the cell line KIS-1, which harbors t(9;14)(p13;q32) and was kindly provided by Dr Hitoshi Ohno (Kyoto University, Japan) and in a patient sample with t(9;14)(p13;q32) that was kindly provided by Dr Herve Avet-Loiseau (Nantes, France). In both cases our probes showed 2 fusion signals of the IgH probe and the PAX-5 BAC clone, on both derivative chromosomes in each case, indicating its usefulness for the detection of this translocation.
To detect t(11;18)(q21;q21) we used a set of probes, kindly provided by Dr David James (Mayo Clinic), that span both breakpoints in all reported t(11;18)(q21;q21) translocations and that result in a double fusion, as validated by Remstein et al27 (Figure 2). To screen for possible aneuploidy, we used commercially available centromere enumeration probes (CEPs) for chromosomes 7, 9, 11, 12, 15, and 17 (Vysis). To screen for 6q21 deletions, we used the clone RPCI 91C23 (obtained from Oakland Children's Hospital, Oakland, CA) with simultaneous hybridization for CEP 6. This same cohort of patients has also been studied for deletions at 13q14 and 17p13 by interphase FISH, and the results are compared with those obtained from this study.28 Noncommercial probes were directly labeled using standard nick translation with SpectrumRed or SpectrumGreen (Vysis). Slides and probes were co-denatured for 7 minutes at 80°C, placed in a humidified chamber, and allowed to hybridize in a 37°C oven overnight. Slides were then washed, an antifade mounting medium (Vectashield H-1000) was applied to each, and the slides were coverslipped. For slides not assessed for cytoplasmic immunoglobulin stain, DAPI (Vector Laboratories, Burlingame, CA) was added to the medium. Cytoplasmic staining In all patients (n = 33) for whom cytospin slides were available, we performed cytoplasmic staining of the IgM cytoplasmic protein using an AMCA (7-amino-4-methylcoumarin-3-acetic acid)-conjugated anti-IgM antibody (Vector Laboratories), in a variation of our previously published technique.29 If we could not use the cIg-FISH technique, or if the original cytospin slides were depleted, we used slides made from the cultured and fixed cells. Because of these limitations, these slides were scored on unselected cells without regard to cellular morphology.Scoring statistics Using a Leica epifluorescence microscope with fluorescein isothiocyanate, Texas red, and a DAPI filter, we attempted to score at least 100 clonal cells from each patient. In the cytospin slides, only cells identified by morphologic features or cytoplasmic anti-IgM staining were scored. Fusion of probe signals (colocalization) was defined as 2 probe signals making contact (Figure 2). Break-apart of probe signals (segregation) was defined by a distance of more than 3 signal widths between 2 differently labeled probe signals. A sample was said to have an abnormal pattern if the percentage of cells exceeded the mean percentage background level plus 3 SD found for normal cells. The mean percentage background was determined in normal and abnormal samples, and at least 1000 cells were scored for each set of probes.
Southern blot analysis To detect IgH rearrangements we conducted a pilot study using Southern blot analysis, but we did not find evidence of legitimate or illegitimate IgH switch recombination fragments. In all cases the presence of clonal cell DNA was confirmed by the finding of clonal rearrangements by hybridization with a JH probe, but there was complete concordance of the 5'Sµ and the 3'Sµ bands in sequential hybridizations (n = 12) (Figure 1).Conventional cytogenetics and M-FISH We reviewed the karyotype results of 45 patients using conventional cytogenetic analysis performed for routine clinical purposes. Six patients were excluded because chromosomal analysis was performed at the time of diagnosis of secondary myelodysplasia or leukemia. Abnormal findings on BM examination and resultant abnormal karyotypes were consistent with the diagnoses. Of the remaining 39 patients, 35 had sufficient numbers of metaphases to be evaluable (Table 2). Of these 35 patients, 22 (63%) patients were found to have only normal metaphases, possibly originating from the normal myeloid elements in the BM, including 2 patients whose only abnormality was the loss of chromosome Y. Abnormal metaphases were obtained in 13 (37%) patients, 5 of whom were not previously treated. Structural and numerical chromosomal abnormalities were encountered in the abnormal karyotypes, but usually we observed single abnormalities in an otherwise normal karyotype (Table 2). For the 13 patients with abnormal karyotype, 4 (31%) had recurrent abnormalities of chromosome 13, with del(13)(q14) in 3 and monosomy 13 in one; 2 (15%) had del(6)(q13q21)(q13q25); 1 (8%) had an addition at chromosome 6q27 (add(6)(q27)); 2 (15%) had abnormalities of chromosome 17q25 and 1 (8%) had chromosome 17 monosomy; 2 had del(5q)(13q35) (one with and one without prior therapy); and 4 had diverse abnormalities in chromosome 8 (t(Y;8)(q11;p23), i8(q10), der(8;9)(q10;q10), and 8). As a follow-up
to conventional cytogenetics in 5 of 13 patients with abnormal
karyotype, M-FISH studies identified karyotypic abnormalities in 2 patients but provided no additional information in 3 patients
(Table 2).
Interphase FISH Of the 31 patients studied, 30 had no evidence of IgH translocations by the VH/CH strategy, and in one patient 37% of cells harbored the abnormality. In addition, in none of these 31 patients could we find fusion signals indicative of t(9;14)(p13;q32) (Figure 2). When we extended our study to include 18 additional patients in whom we had CC cell suspension to study, we found that none of them had evidence of IgH translocations by the VH/CH break-apart strategy (n = 17) or the t(9;14)(p13;q32) translocation by the fusion strategy (n = 18). One patient had 3 or more signals from the PAX-5 BAC clone in 69% of the cells but no fusions (17% with 3 signals, and 52% with 4 signals). This same patient had evidence of trisomy for chromosome 9 in the metaphase analysis. None of the 24 patients studied had evidence of t(11;18)(q21;q21) as detected by FISH.To screen for possible aneuploidy, tumor cells from 15 patients were studied by interphase FISH using CEP probes for chromosomes 7, 9, 11, 12, 15, and 17. With the exception of one patient, all chromosomes were normal; one patient had only one signal for chromosome 9, indicative of monosomy in 26% of the clonal cells. Although some patients showed abnormalities in the karyotype consistent with aneuploidy (Table 2), interphase FISH showed that this is the exception, and patients with karyotype abnormalities likely have aggressive variants of the disease. There is probably an inherent bias in the patient population that is subjected to scrutiny of karyotype analysis (ie, worsening disease). In addition, it is highly likely, as in multiple myeloma, that the ability to obtain informative metaphases is closely related to tumor burden and proliferative activity.30 Deletions of 6q We also tested 24 patients (in whom we had metaphase culture slides but could not perform cIg-FISH assay) for evidence of 6q21 deletions and found that 10 (42%) had abnormalities in more than 25% of the cells. In addition 5 (21%) more patients had between 10% and 25% abnormal cells. Because in many of these latter patients the percentage of clonal involvement of the bone marrow was between 10% and 20%, it is conceivable that they could also have had deletions at this site. Therefore, between 42% and 63% of patients had deletions of 6q21.Correlation between interphase FISH and karyotype analysis Three patients had chromosome abnormalities detected by metaphase analysis; we also performed specific interphase FISH. In one patient13 was confirmed by FISH, and in another patient the deletion of 17p
was confirmed by FISH. In yet another patient, del(6)(q13q25) was
detected by our FISH probe.
In this study we have found that clonal cells from patients with
clinically defined WM do not have the t(9;14)(p13;q32) translocation, as previously found in 50% of patients with LPL but without
paraproteinemia. Thus, WM, a type of LPL with IgM paraproteinemia, and
LPL with no paraproteinemia differ not only in phenotype but also in
the presence of the t(9;14)(p13;q32) translocation. This result is consistent with the hypothesis that the predicted biologic features of
B-cell clones with PAX-5 up-regulation are incompatible with the phenotype of WM.17-19 Notably, BSAP is absent in
plasma cells, and the overexpression of PAX-5/BSAP abrogates
the production of the J peptide.13 This protein is an
integral component of the IgM pentamers, which give rise to the
hyperviscosity of patients with WM.31 In addition,
PAX-5/BSAP negatively regulates the 3' Our finding that IgH translocations are not present in WM is consistent with the previously reported karyotypic abnormalities in these patients. In a series of 45 patients with WM reported by Louviaux et al,34 12 patients had abnormal metaphases, but no abnormalities of chromosome 14 were noted. In our review of the cytogenetic database at the Mayo Clinic, which we report here, we were unable to find any patient with 14q32 abnormalities. Unlike multiple myeloma20,35,36 and low-grade lymphomas,37,38 14q32 translocations appear not to be initiation events for disease pathogenesis in WM. Scattered reports in the literature describe patients with WM and 14q32 translocations, including t(8;14)(q24;q32).39-42 Some of these reports are confounded either by the lack of consistent clinical features associated with a diagnosis of WM or by the samples originating from pleural effusions and, thus, of unknown relation to the original clone.39-42 There are rare reports of WM patients in whom tumor cells had a t(11;18)(q21;q21) translocation,43,44 an abnormality also seen in approximately 20% of extranodal marginal zone lymphomas.45 Because we have been unable to detect this translocation in the tumor cells of 24 WM patients, it must be, at best, rare. Despite an apparent paucity of structural and numeric karyotypic abnormalities in tumor cells from our cohort of WM patients, we did identify a high prevalence of deletions of the long arm of chromosome 6. This is consistent with what has been previously reported in selected patients with WM46,47 and other B-cell neoplasias in which abnormalities in this region are common.48-50 This is also consistent with findings from our karyotype analysis that 3 patients had 6q deletions. Further work is under way to characterize the area of minimal deletion and the search for putative genes involved in disease pathogenesis. We suspect that the inactivation of a tumor suppressor gene at this locus, as is seen in other B-cell neoplasias, is likely to be of importance for clone immortalization. Our data suggest less genomic instability in WM than in multiple
myeloma, as determined by metaphase analyses for structural and numeric
abnormalities and interphase FISH for numeric abnormalities. Moreover,
we have recently reported that, unlike multiple myeloma and B-cell
chronic lymphocytic leukemia (B-CLL), deletions of 13q14 are rarely
observed at the time of diagnosis.28 As are deletions of
17p13.1 (see below), 13q14 deletions are seen mostly in advanced stages
and in clonal cells in a small fraction (approximately 15%) of
patients with WM at the time of diagnosis.28 Therefore, we
can conclude that WM is clearly different from myeloma in that it lacks
IgH translocations, has infrequent deletions of 13q14, and has limited
numeric and structural karyotypic abnormalities. WM is also different
from B-CLL in the lower incidence of 13q14 deletions (seen in
approximately 50% of patients with B-CLL) and the low prevalence of
trisomy 12 (seen in approximately 15% of patients with
B-CLL).51 WM is also different from extranodal marginal
zone lymphomas in that we could not detect any patient with
t(11;18)(q21;q21). The lack of IgH translocations also differentiates WM from many kinds of B lymphomas (follicular, mantle cell, diffuse large cell).52 We thus conclude that the biologic nature
of WM is different from that of multiple myeloma and most lymphomas, but it appears similar to postgerminal center B-CLL (Table
3).
The normal counterpart of the malignant cell in WM might be a postgerminal center IgM memory B cell that has undergone somatic hypermutation but has failed to undergo isotype class switching. Our Southern blot results, failing to identify legitimate or illegitimate IgH switch recombination rearrangements indicative of isotype class switching, and the presence of the IgM paraproteinemia are consistent with this. In addition, individual WM tumor cells show heterogeneous morphologic features consistent with variable differentiation from a B cell to a plasma cell but failure to fully differentiate into plasma cells. We speculate that the presumed genetic event(s) responsible for generating an immortalized clone of tumor cells in WM are directly or indirectly responsible for the failure of isotype class switching and the lack of differentiation to plasma cells. With the exception of frequent deletions of the long arm of chromosome 6, our knowledge of specific primary or secondary genetic events in WM remains elusive.
Submitted August 31, 2001; accepted June 7, 2002.
R.F. is supported by a research grant from the International Waldenström Macroglobulinemia Foundation and is a Leukemia and Lymphoma Society Translational Research Awardee. R.F. and P.R.G. are supported by the CI-5 Cancer Research Fund-Lilly Clinical Investigator Award of the Damon Runyon-Walter Winchell Foundation. Supported in part by Public Health Service grant R01 CA83724-01 from the National Cancer Institute (R.F.). P.R.G. and R.A.K. are supported in part by research grant P01 CA62242 from the National Cancer Institute. P.R.G. is supported by the ECOG grant CA21115-25C from the National Cancer Institute.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Rafael Fonseca, Department of Hematology and Internal Medicine, Stabile 628, Mayo Clinic, Rochester, MN 55905; e-mail: fonseca.rafael{at}mayo.edu.
1.
Dimopoulos MA, Panayiotidis P, Moulopoulos LA, Sfikakis P, Dalakas M.
Waldenström's macroglobulinemia: clinical features, complications, and management [review].
J Clin Oncol.
2000;18:214-226
2.
Gertz MA, Fonseca R, Rajkumar SV.
Waldenström's macroglobulinemia [review].
Oncologist.
2000;5:63-67
3.
Harris NL, Jaffe ES, Stein H, et al.
A revised European-American classification of lymphoid neoplasms: a proposal from the International Lymphoma Study Group [review].
Blood.
1994;84:1361-1392
4.
Offit K, Parsa NZ, Filippa D, Jhanwar SC, Chaganti RS.
t(9;14)(p13;q32) denotes a subset of low-grade non-Hodgkin's lymphoma with plasmacytoid differentiation.
Blood.
1992;80:2594-2599 5. Stapleton P, Weith A, Urbanek P, Kozmik Z, Busslinger M. Chromosomal localization of seven PAX genes and cloning of a novel family member, PAX-9. Nat Genet. 1993;3:292-298[CrossRef][Medline] [Order article via Infotrieve].
6.
Ohno H, Furukawa T, Fukuhara S, et al.
Molecular analysis of a chromosomal translocation, t(9;14)(p13;q32), in a diffuse large-cell lymphoma cell line expressing the Ki-1 antigen.
Proc Natl Acad Sci U S A.
1990;87:628-632
7.
Iida S, Rao PH, Nallasivam P, et al.
The t(9;14)(p13;q32) chromosomal translocation associated with lymphoplasmacytoid lymphoma involves the PAX-5 gene.
Blood.
1996;88:4110-4117 8. Iida S, Rao PH, Ueda R, Chaganti RS, Dalla-Favera R. Chromosomal rearrangement of the PAX-5 locus in lymphoplasmacytic lymphoma with t(9;14)(p13;q32) [review]. Leuk Lymphoma. 1999;34:25-33[Medline] [Order article via Infotrieve]. 9. Ohno H, Ueda C, Akasaka T. The t(9;14)(p13;q32) translocation in B-cell non-Hodgkin's lymphoma [review]. Leuk Lymphoma. 2000;36:435-445[Medline] [Order article via Infotrieve]. 10. Barr FG. Chromosomal translocations involving paired box transcription factors in human cancer [review]. Int J Biochem Cell Biol. 1997;29:1449-1461[CrossRef][Medline] [Order article via Infotrieve].
11.
Wakatsuki Y, Neurath MF, Max EE, Strober W.
The B cell-specific transcription factor BSAP regulates B cell proliferation.
J Exp Med.
1994;179:1099-1108 12. Urbanek P, Wang ZQ, Fetka I, Wagner EF, Busslinger M. Complete block of early B cell differentiation and altered patterning of the posterior midbrain in mice lacking Pax5/BSAP. Cell. 1994;79:901-912[CrossRef][Medline] [Order article via Infotrieve].
13.
Morrison AM, Jager U, Chott A, Schebesta M, Haas OA, Busslinger M.
Deregulated PAX-5 transcription from a translocated IgH promoter in marginal zone lymphoma.
Blood.
1998;92:3865-3878
14.
Adams B, Dorfler P, Aguzzi A, et al.
Pax-5 encodes the transcription factor BSAP and is expressed in B lymphocytes, the developing CNS, and adult testis.
Genes Dev.
1992;6:1589-1607
15.
Krenacs L, Himmelmann AW, Quintanilla-Martinez L, et al.
Transcription factor B-cell-specific activator protein (BSAP) is differentially expressed in B cells and in subsets of B-cell lymphomas.
Blood.
1998;92:1308-1316 16. Busslinger M, Urbanek P. The role of BSAP (Pax-5) in B-cell development [review]. Curr Opin Genet Dev. 1995;5:595-601[CrossRef][Medline] [Order article via Infotrieve].
17.
Neurath MF, Max EE, Strober W.
Pax5 (BSAP) regulates the murine immunoglobulin 3' alpha enhancer by suppressing binding of NF-alpha P, a protein that controls heavy chain transcription.
Proc Natl Acad Sci U S A.
1995;92:5336-5340 18. Max EE, Wakatsuki Y, Neurath MF, Strober W. The role of BSAP in immunoglobulin isotype switching and B-cell proliferation. Curr Topics Microbiol Immunol. 1995;194:449-458[Medline] [Order article via Infotrieve]. 19. Neurath MF, Stuber ER, Strober W. BSAP: a key regulator of B-cell development and differentiation [review]. Immunol Today. 1995;16:564-569[CrossRef][Medline] [Order article via Infotrieve].
20.
Bergsagel PL, Chesi M, Nardini E, Brents LA, Kirby SL, Kuehl WM.
Promiscuous translocations into immunoglobulin heavy chain switch regions in multiple myeloma.
Proc Natl Acad Sci U S A.
1996;93:13931-13936 21. Owen R, Parapia L, Richards S, et al. Should Waldenström's macroglobulinemia be considered part of the spectrum of marginal zone lymphoma [abstract]? Blood. 1999;94(suppl 1):385. 22. Owen R, Johnson S, Morgan G. Waldenström's macroglobulinemia: laboratory diagnosis and treatment. Hematol Oncol. 2000;18:41-49[CrossRef][Medline] [Order article via Infotrieve]. 23. Spurbeck JL, Carlson RO, Allen JE, Dewald GW. Culturing and robotic harvesting of bone marrow, lymph nodes, peripheral blood, fibroblasts, and solid tumors with in situ techniques. Cancer Genet Cytogenet. 1988;32:59-66[CrossRef][Medline] [Order article via Infotrieve]. 24. Jalal SM, Law ME, Stamberg J, et al. Detection of diagnostically critical, often hidden, anomalies in complex karyotypes of haematological disorders using multicolour fluorescence in situ hybridization. Br J Haematol. 2001;112:975-980[CrossRef][Medline] [Order article via Infotrieve]. 25. Schrock E, Veldman T, Padilla-Nash H, et al. Spectral karyotyping refines cytogenetic diagnostics of constitutional chromosomal abnormalities. Hum Genet. 1997;101:255-262[CrossRef][Medline] [Order article via Infotrieve]. 26. Gabrea A, Bergsagel PL, Chesi M, Shou Y, Kuehl WM. Insertion of excised IgH switch sequences causes overexpression of cyclin D1 in a myeloma tumor cell. Mol Cell. 1999;3:119-123[CrossRef][Medline] [Order article via Infotrieve].
27.
Remstein ED, James CD, Kurtin PJ.
Incidence and subtype specificity of API2-MALT1 fusion translocations in extranodal, nodal, and splenic marginal zone lymphomas.
Am J Pathol.
2000;156:1183-1188 28. Schop R, Jalal S, Van Wier S, et al. Deletions of 17p13.1 and 13q14 are uncommon in Waldenström macroglobulinemia clonal cells and mostly seen at the time of disease progression. Cancer Genet Cytogenet. 2002;132:55-60[CrossRef][Medline] [Order article via Infotrieve]. 29. Ahmann GJ, Jalal SM, Juneau AL, et al. A novel three-color, clone-specific fluorescence in situ hybridization procedure for monoclonal gammopathies. Cancer Genet Cytogenet. 1998;101:7-11[CrossRef][Medline] [Order article via Infotrieve]. 30. Rajkumar SV, Fonseca R, Dewald GW, et al. Cytogenetic abnormalities correlate with the plasma cell labeling index and extent of bone marrow involvement in myeloma. Cancer Genet Cytogenet. 1999;113:73-77[CrossRef][Medline] [Order article via Infotrieve]. 31. Gertz MA, Kyle RA. Hyperviscosity syndrome. J Intens Care. 1995;10:128-141.
32.
Singh M, Birshtein BK.
NF-HB (BSAP) is a repressor of the murine immunoglobulin heavy-chain 3' alpha enhancer at early stages of B-cell differentiation.
Mol Cell Biol.
1993;13:3611-3622 33. Neurath MF, Strober W, Wakatsuki Y. The murine Ig 3' alpha enhancer is a target site with repressor function for the B cell lineage-specific transcription factor BSAP (NF-HB, S alpha-BP). J Immunol. 1994;153:730-742[Abstract]. 34. Louviaux I, Michaux L, Hagemeijer A, et al. Cytogenetic abnormalities in Waldenström's disease (WD): a single centre study of 45 cases [abstract]. Vol 92. In: Proceedings of the Annual Meeting of the American Society of Hematology.; 1998:184b.
35.
Hallek M, Bergsagel PL, Anderson KC.
Multiple myeloma: increasing evidence for a multistep transformation process.
Blood.
1998;91:3-21 36. Bergsagel PL, Nardini E, Brents L, Chesi M, Kuehl WM. IgH translocations in multiple myeloma: a nearly universal event that rarely involves c-myc. Curr Top Microbiol Immunol. 1997;224:283-287[Medline] [Order article via Infotrieve].
37.
Levine EG, Arthur DC, Frizzera G, Peterson BA, Hurd DD, Bloomfield CD.
There are differences in cytogenetic abnormalities among histologic subtypes of the non-Hodgkin's lymphomas.
Blood.
1985;66:1414-1422 38. Yunis JJ, Oken MM, Kaplan ME, Ensrud KM, Howe RR, Theologides A. Distinctive chromosomal abnormalities in histologic subtypes of non-Hodgkin's lymphoma. N Engl J Med. 1982;307:1231-1236[Abstract]. 39. Chong YY, Lau LC, Lui WO, et al. A case of t(8;14) with total and partial trisomy 3 in Waldenström macroglobulinemia. Cancer Genet Cytogenet. 1998;103:65-67[CrossRef][Medline] [Order article via Infotrieve]. 40. San Roman C, Ferro T, Guzman M, Odriozola J. Clonal abnormalities in patients with Waldenström's macroglobulinemia with special reference to a Burkitt-type t(8;14). Cancer Genet Cytogenet. 1985;18:155-158[CrossRef][Medline] [Order article via Infotrieve].
41.
Nishida K, Taniwaki M, Misawa S, Abe T.
Nonrandom rearrangement of chromosome 14 at band q32.33 in human lymphoid malignancies with mature B-cell phenotype.
Cancer Res.
1989;49:1275-1281 42. Calasanz MJ, Cigudosa JC, Odero MD, et al. Cytogenetic analysis of 280 patients with multiple myeloma and related disorders: primary breakpoints and clinical correlations. Genes Chromosomes Cancer. 1997;18:84-93[CrossRef][Medline] [Order article via Infotrieve]. 43. Hirase N, Miyamura T, Ishikura H, et al. Primary macroglobulinemia with t(11;18) (q21;q21) [review]. Rinsho Ketsueki. 1996;37:340-345[Medline] [Order article via Infotrieve]. 44. Hirase N, Yufu Y, Abe Y, et al. Primary macroglobulinemia with t(11;18)(q21;q21). Cancer Genet Cytogenet. 2000;117:113-117[CrossRef][Medline] [Order article via Infotrieve]. 45. Dierlamm J, Wlodarska I, Michaux L, et al. Genetic abnormalities in marginal zone B-cell lymphoma [review]. Hematol Oncol. 2000;18:1-13[CrossRef][Medline] [Order article via Infotrieve]. 46. Wong KF, So CC. Waldenström macroglobulinemia with karyotypic aberrations involving both homologous 6q. Cancer Genet Cytogenet. 2001;124:137-139[CrossRef][Medline] [Order article via Infotrieve].
47.
Mansoor A, Medeiros LJ, Weber DM, et al.
Cytogenetic findings in lymphoplasmacytic lymphoma/Waldenström macroglobulinemia: chromosomal abnormalities are associated with the polymorphous subtype and an aggressive clinical course.
Am J Clin Pathol.
2001;116:543-549 48. Parsa NZ, Gaidano G, Mukherjee AB, et al. Cytogenetic and molecular analysis of 6q deletions in Burkitt's lymphoma cell lines. Genes Chromosomes Cancer. 1994;9:13-18[Medline] [Order article via Infotrieve].
49.
Offit K, Parsa NZ, Gaidano G, et al.
6q deletions define distinct clinico-pathologic subsets of non-Hodgkin's lymphoma.
Blood.
1993;82:2157-2162
50.
Gaidano G, Hauptschein RS, Parsa NZ, et al.
Deletions involving two distinct regions of 6q in B-cell non-Hodgkin lymphoma.
Blood.
1992;80:1781-1787
51.
Dohner H, Stilgenbauer S, Benner A, et al.
Genomic aberrations and survival in chronic lymphocytic leukemia.
N Engl J Med.
2000;343:1910-1916
52.
Willis TG, Dyer MJS.
The role of immunoglobulin translocations in the pathogenesis of B-cell malignancies.
Blood.
2000;96:808-822
© 2002 by The American Society of Hematology.
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
|
 |
|
  S. P. Treon How I treat Waldenstrom macroglobulinemia Blood, September 17, 2009; 114(12): 2375 - 2385. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Landgren, S. Y. Kristinsson, L. R. Goldin, N. E. Caporaso, C. Blimark, U.-H. Mellqvist, A. Wahlin, M. Bjorkholm, and I. Turesson Risk of plasma cell and lymphoproliferative disorders among 14621 first-degree relatives of 4458 patients with monoclonal gammopathy of undetermined significance in Sweden Blood, July 23, 2009; 114(4): 791 - 795. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. M. Roccaro, A. Sacco, C. Chen, J. Runnels, X. Leleu, F. Azab, A. K. Azab, X. Jia, H. T. Ngo, M. R. Melhem, et al. microRNA expression in the biology, prognosis, and therapy of Waldenstrom macroglobulinemia Blood, April 30, 2009; 113(18): 4391 - 4402. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Braggio, J. J. Keats, X. Leleu, S. Van Wier, V. H. Jimenez-Zepeda, R. Valdez, R. F.J. Schop, T. Price-Troska, K. Henderson, A. Sacco, et al. Identification of Copy Number Abnormalities and Inactivating Mutations in Two Negative Regulators of Nuclear Factor-{kappa}B Signaling Pathways in Waldenstrom's Macroglobulinemia Cancer Res., April 15, 2009; 69(8): 3579 - 3588. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Y. Kristinsson, M. Bjorkholm, L. R. Goldin, M. L. McMaster, I. Turesson, and O. Landgren Risk of lymphoproliferative disorders among first-degree relatives of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia patients: a population-based study in Sweden Blood, October 15, 2008; 112(8): 3052 - 3056. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Quijano, A. Lopez, A. Rasillo, S. Barrena, M. Luz Sanchez, J. Flores, C. Fernandez, J. M. Sayagues, C. S. Osuna, N. Fernandez, et al. Association between the proliferative rate of neoplastic B cells, their maturation stage, and underlying cytogenetic abnormalities in B-cell chronic lymphoproliferative disorders: analysis of a series of 432 patients Blood, May 15, 2008; 111(10): 5130 - 5141. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Vijay and M. A. Gertz Waldenstrom macroglobulinemia Blood, June 15, 2007; 109(12): 5096 - 5103. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. J. Chng, R. F. Schop, T. Price-Troska, I. Ghobrial, N. Kay, D. F. Jelinek, M. A. Gertz, A. Dispenzieri, M. Lacy, R. A. Kyle, et al. Gene-expression profiling of Waldenstrom macroglobulinemia reveals a phenotype more similar to chronic lymphocytic leukemia than multiple myeloma Blood, October 15, 2006; 108(8): 2755 - 2763. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. P. Treon, Z. R. Hunter, A. Aggarwal, E. P. Ewen, S. Masota, C. Lee, D. D. Santos, E. Hatjiharissi, L. Xu, X. Leleu, et al. Characterization of familial Waldenstrom's macroglobulinemia Ann. Onc., March 1, 2006; 17(3): 488 - 494. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Fonseca A "not so micro" model for a "macro" problem Blood, August 15, 2005; 106(4): 1143 - 1144. [Full Text] [PDF]
|
|
|
|
 |
|
 T. I. George, J. E. Wrede, C. D. Bangs, A. M. Cherry, R. A. Warnke, and D. A. Arber Low-Grade B-Cell Lymphomas With Plasmacytic Differentiation Lack PAX5 Gene Rearrangements J. Mol. Diagn., August 1, 2005; 7(3): 346 - 351. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. A. Gertz, G. Merlini, and S. P. Treon Amyloidosis and Waldenstrom's Macroglobulinemia Hematology, January 1, 2004; 2004(1): 257 - 282. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
 |
 |
|||||||
 |
 |
 |
 |
 |
 |
 |
 |
||
| Copyright © 2002 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
Top
Abstract
Introduction
promoter and the IgH 3'
enhancer,
with consequent down-regulation of IgH transcription.
including the t(11;18)(q21;q21) translocation or
abnormalities of 6q21
HindIII markers, with the 9.4-kb marker in
each lane but the 6.6-kb marker included for panel A. Panel A shows the
genomic DNA probes with a JH oligonucleotide probe and the
corresponding clonal bands (variable size) and germ line fragments
(approximately 10 kb). Panel B shows the constant size of the 5' Sµ
probe indicating lack of legitimate or illegitimate rearrangements. The
total amount of genomic DNA loaded was 2.5 µg per lane.
