C-myc Gene Rearrangement Research Articles

Diffuse-Large-B-Cell Lymphoma Patients with Secretory IgM Monoclonal Component Are A Very Poor Prognostic Subset.

Abstract 2659 Background:Gene-expression-profiling defined at least two main groups within Diffuse-large-B-cell Lymphoma (DLBCL) patients who have substantially different outcomes: Activated-B-cell (ABC-type) and Germinal-Center-B-cell (GCB-type). The translation of gene-expression-profiling arrays into robust algorithm useful for clinical purposes is still in progress. The detection of IgM monoclonal component (IgM MC) in DLBCL has been previously described in a few reports, mainly because it was associated with autoimmune hemolytic anaemia. To our knowledge this is the first report describing the incidence and prognosis of a series of DLBCL with IgM MC. Aims:In this report we compared clinical and biological features of DLBCL patients with and without secretory IgM MC . Patients & Methods:Within a consecutive series of 132 patients, diagnosed between September 2004 and April 2012 with conventional DLBCL, 16 cases (12%) with a IgM MC were identified. We selected a set of 95 consecutive DLBCL patients, treated with 6–8 cycles of RCHOP-like for comparison of histological features and survival. Only cases with a follow up time >24 months were included, unless a DLBCL–related event (i.e. primary refractoriness or relapse) had occurred earlier. Biological material was obtained after receiving patient’s consent. This study was approved by our Institutional-Review-Board. Immunohistochemistry and FISH:Paraffin sections were immunostained for CD3, CD5, CD20, CD10, CD30, CD79a, CD138, ALK-1, MUM1, BCL2, BCL6, IgM, Kappa and Lambda immunoglobulin light chains, using an automated immunostainer (DAKO, Denmark). The Hans algorithm was used in order to classify cases as GCB-type and non GCB-type. FISH with Vysis break-apart probe was used to assess c-MYC gene abnormalities in tissue sections (Abbott Molecular Inc. US). Statistics:univariate comparisons between groups were carried out with appropriate non parametric test. Survival analyses were done by the Kaplan-Meier method, the analyses of factor predicting survival were carried out by the log-rank test. Cox's regression was used for multivariate analyses. The SPSS19 package (SPSS Inc.Chicago IL) was used for elaborations. Histology, immunohistochemistry and FISH Results:In 14 out of 16 cases (87.5%) the IgM MC was related to the DLBCL clone. This was ascertained by immunostaining of cytoplasm for IgM, Kappa and Lambda immunoglobulin light chains. All the 14 cases were classified as non GCB-type. FISH analysis detected no c-MYC gene rearrangements in all the cases. Clinical Results:The incidence of bone marrow involvement, two or more extranodal sites, female sex, IPI score 3–5 and failure to achieve CR on RCHOP treatment were significantly more frequent in the IgM MC group. Noteworthy four out of 14 patients had central nervous system involvement at diagnosis or at relapse. All but one, with a previous diagnosis of marginal zone lymphoma, were de novo DLBCL. Twelve patients (85.7%) presented a DLBCL related event compared to 35 patients (37%) without IgM MC (p=.001). Seven patients (50%) died with primary refractory or relapsed-chemoresistant disease, another one died of an adverse event during chemotherapy. Two are alive on salvage treatment, two are in PFS at +30 and +13 months after salvage treatment with Bortezomib-RDHAP followed by high dose therapy. Only two patients are in PFS after first line RCHOP at +56 and +29 months respectively. Survival analysis:The estimated two-year EFS, PFS and OS were significantly worst for IgM MC group (22% Vs 70%, p<.0001; 22% Vs 75%, p<.0001; 50% Vs 85%, p=.011, respectively). In multivariate analysis IgM MC was the only significant factor for EFS (p=.001; 0.194 CI:0.089–0.57) while for PFS were both significant the IgM-MC (p=0.014; 0.360; 5%CI: 0,159–0,815) and the IPI-score 3–5 (p=.002; 0.186; 95% CI 0.063–0.552). Conclusions:A secretory IgM MC related to the neoplastic clone, was detected in more than 10% of newly diagnosed DLBCL. IgM MC in DLBCL might be easily missed, given its negligible entity, the rarity of associated clinical signs and the rapid MC fading during treatment. This group showed homogeneous morphologic and immunohistochemical features consistent with the non GCB-type. FISH analysis was negative for c-MYC gene rearrangements. IgM MC in DLBCL patients, might be related to a very poor non GCB-type subset who should be given upfront intensified therapies. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

P53 deletion is independently associated with increased age and decreased survival in a cohort of Chinese patients with diffuse large B-cell lymphoma

The purpose of the study was to compare cytogenetic profiles and survivals between elderly and non-elderly Chinese patients with diffuse large B-cell lymphoma (DLBCL). We identified 50 patients with DLBCL and divided them by age into elderly (≥60 years) and non-elderly (< 60 years) groups. We detected deletion of P53 or translocations in Bcl-2, Bcl-6 or c-myc genes by fluorescence in situ hybridization (FISH). P53 deletion was significantly more common in elderly versus non-elderly patients (62% vs. 17%, respectively, p=0.001) There were no significant differences in rates of Bcl-2, Bcl-6 or c-myc gene rearrangements between elderly and non-elderly patients (p>0.25 for each). Median survival was significantly longer in non-elderly compared to elderly patients. P53 deletion was an independent prognostic factor for decreased survival in patients with DLBCL, independent of age. In conclusion, P53 deletion as detected by FISH is associated with both increased age and decreased survival in Chinese patients with DLBCL. This association with decreased survival is at least partly independent of age. P53 deletion could serve as a prognostic factor in DLBCL independent of or in combination with age.

Burkitt Lymphoma

Burkitt lymphoma (BL) is a B-cell lymphoma with a germinal center phenotype that has a very rapid doubling time, approximately 24 hours, for which early and accurate diagnosis is critical. There are 3 epidemiologic classifications (endemic, sporadic, and immunodeficiency associated); however, all 3 have indistinguishable histopathology and immunohistochemistry. The role of genetics is critical to detect a translocation of the c-MYC gene. c-MYC gene rearrangements are seen not only in BL but also diffuse large B-cell lymphoma and B-cell lymphoma, unclassifiable, with features intermediate between diffuse large B-cell lymphoma and BL. Treatments for BL have improved drastically during the last decade, primarily with the implementation of pediatric regimens in the adult population. The role of the pathologist is critical in the timely implementation of treatment of adequate intensity. As with many other hematopoietic malignancies, refractoriness to initial therapy is associated with a poor prognosis. This review addresses the pathology, clinical features, histogenesis, and molecular findings of BL and its differential diagnosis.

The Germinal Center/Activated B-Cell Subclassification Has a Prognostic Impact for Response to Salvage Therapy in Relapsed/Refractory Diffuse Large B-Cell Lymphoma: A Bio-CORAL Study

To evaluate the prognostic value of the cell of origin (COO) in patients with relapsed/refractory diffuse large B-cell lymphoma (DLBLC), prospectively treated by rituximab, dexamethasone, high-dose cytarabine, and cisplatin (R-DHAP) versus rituximab, ifosfamide, carboplatin, and etoposide and followed by intensive therapy plus autologous stem-cell transplantation on the Collaborative Trial in Relapsed Aggressive Lymphoma (CORAL) trial. Among the 396 patients included on the trial, histologic material was available for a total of 249 patients at diagnosis (n = 189 patients) and/or at relapse (n = 147 patients), which included 87 matched pairs. The patient data were analyzed by immunochemistry for CD10, BCL6, MUM1, FOXP1, and BCL2 expression and by fluorescent in situ hybridization for BCL2, BCL6 and c-MYC breakpoints. The correlation with survival data was performed by using the log-rank test and the Cox model. Characteristics of immunophenotype and chromosomal abnormalities were statistically highly concordant in the matched biopsies. In univariate analysis, the presence of c-MYC gene rearrangement was the only parameter to be significantly correlated with a worse progression-free survival (PFS; P = .02) and a worse overall survival (P = .04). When treatment interaction was tested, the germinal center B (GCB) -like DLBCL that was based on the algorithm by Hans was significantly associated with a better PFS in the R-DHAP arm. In multivariate analysis, independent prognostic relevance was found for the GCB/non-GCB the Hans phenotype interaction treatment (P = .04), prior rituximab exposure (P = .0052), secondary age-adjusted International Prognostic Index (P = .039), and FoxP1 expression (P = .047). Confirmation was obtained by gene expression profiling in a subset of 39 patients. COO remains a major and independent factor in relapsed/refractory DLBCL, with a better response to R-DHAP in GCB-like DLBCL. This needs confirmation by a prospective study.

Clinical spectrum and prognosis of follicular lymphoma with blastoid transformation: case series and a review of the literature

Follicular lymphoma (FL) with blastoid transformation (FL-BT) is a rare entity. FL-BT has a dismal prognosis due to intrinsic biological characteristics, lack of evidence-based therapeutic guidelines, and exclusion from clinical trials. In order to understand the natural course of the disease and reconcile treatment strategies in patients with FL-BT, we reviewed available data. Thirty patients were identified, three from our institution (Roswell Park Cancer Institute) and 27 patients from peer-reviewed literature. Demographic, clinical, treatment, pathological, and cytogenetic characteristics were described. Most of the patients (90%) were treated with chemotherapy at transformation. Complete and partial responses to treatment were observed in 38.9% and 22.3% of the patients, respectively. The median overall survival for the entire group of patients was 11 months (range 0.8-30 months); half of the patients were found to have concurrent genetic alterations in the BCL-2 and C-MYC gene rearrangements (double hit). FL-BT with a double hit had a shorter median overall survival when compared to nondouble hit FL-BT (7 months vs. 26 months, respectively, P = 0.005). In our experience, FL-BT is a rare and grave clonal evolution in FL characterized by an aggressive clinical course, inferior response to standard chemoimmunotherapy, and poor clinical outcome. C-MYC rearrangements are common in FL-BT and may play a role in the progression from FL to FL-BT or the poor clinical outcomes observed. Periodic updates from academic institutions treating FL-BT may improve patient care.

Molecular genetic features of sporadic Burkitt's lymphoma in children

To investigate the molecular genetic features and diagnostic aspects of sporadic Burkitt's lymphoma (BL) in children. Tissue microarray was constructed to include 64 cases of pediatric BL and 6 cases of pediatric diffuse large B-cell lymphoma (DLBCL). Immunohistochemistry and fluorescence in-situ hybridization for c-myc, bcl-2, bcl-6, IgH, myc/IgH and bcl-2/IgH gene were performed. Cases of pediatric Burkitt's lymphomas were subclassified into three groups based on their cellular orgins: the germinal center (GC) group, the late-germinal center (late-GC) group and the post-germinal center (post-GC) group. Among 64 Burkitt's lymphomas studied, expression of CD20, CD10, bcl-6, bcl-2 and MUM1 by immunohistochemistry were 100% (64 cases), 98.4% (63 cases), 96.9% (62 cases), 0 (0 cases) and 23.4% (15 cases), respectively. Various gene rearrangements were found involving the c-myc 93.1% (54/58 cases) and IgH 82.8% (48/58 cases). Detailed rearrangements are as follows: 46 cases (85.2%) myc/IgH gene translocation along with c-myc and IgH gene rearrangement; 4 cases (7.4%) c-myc gene rearrangement without IgH and myc/IgH abnormality; 4 cases (7.4%) without c-myc, IgH or myc/IgH gene rearrangement. No case showed bcl-2 gene abnormality (100%). Fifty nine cases showed normal bcl-6 gene status. One case had bcl-6 gene rearrangement and amplification with the pathologic and immunophenotypic characteristics of BL, leading to a revised pathological diagnosis of B-cell lymphoma, unclassifiable with features intermediate between diffuse large B-cell lymphoma and Burkitt's lymphoma (DLBCL/BL). Two cases showed c-myc gene rearrangement. Two cases showed bcl-6 gene amplification and 6 DLBCL cases had a normal status of bcl-2/IgH. A majority of pediatric sporadic BL arise from the germinal center B cells, most of which have c-myc gene rearrangement. It is useful to distinguish BL and DLBCL by multiple genes detection.

Transformation of Follicular Lymphoma to Plasmablastic Lymphoma With c-myc Gene Rearrangement

Follicular lymphoma (FL) is an indolent lymphoma that transforms to high-grade lymphoma, mostly diffuse large B-cell lymphoma, in about a third of patients. We present the first report of a case of FL that transformed to plasmablastic lymphoma (PBL). Clonal transformation of the FL to PBL was evidenced by identical IGH/BCL2 gene rearrangements and VDJ gene usage in rearranged IGH genes. IGH/ BCL2 translocation was retained in the PBL, which also acquired c-myc gene rearrangement. Genealogic analysis based on somatic hypermutation of the rearranged IGH genes of both FL and PBL suggests that transformation of the FL to PBL occurred most likely by divergent evolution from a common progenitor cell rather than direct evolution from the FL clone. Our study of this unusual case expands the histologic spectrum of FL transformation and increases our understanding of the pathogenetic mechanisms of transformation of indolent lymphomas to aggressive lymphomas.

Comparative Genomic Identification of Unique Signal Transduction Pathways and Targets In Pediatric Burkitt Lymphoma (PBL)

AbstractAbstract 631 Background:The prognosis of PBL has dramatically improved over the past 25 years (> 80% 5 yrs EFS) (Cairo et al BJH 2009). This success, however, has come at the cost of the high toxicity associated with intensive multi-agent chemotherapy. A need to identify less toxic targeted therapy exists. Our recent studies show that while almost 100% of childhood BL (238 children) is characterized by an 8q24 (c-myc) gene rearrangement, 70% of the patients also had a secondary chromosomal aberration (Cairo et al Blood 2007), and those children with BL who had a 13q abnormality had a significantly poorer outcome (20% reduction in EFS; Poirel/Cairo et al Leukemia, 2009). Loss of 13q14.3 was associated with a significant decrease in OS (Nelson/Cairo/Perkins/Sanger et al BJH 2010). DLEU1, proven as a Burkitt specific classifier and a cMYC target gene (Dave/Staudt et al NEJM 2006), is located within the region of 13q14.3. DLEU1 is recognized to interact with c-Myc, TUBB2C, UBR1, and other proteins. When DLEU1 is down-regulated by a DLEU1 siRNA, the spontaneous apoptotic rate was decreased with concomitant significantly reduced levels of UBR1 and TUBB2C gene expression (Day/Cairo et al SIOP 2008). Our hypothesis is that deletion of 13q14.3, which contains DLEU1, in pediatric BL may confer a phenotype of chemoimmunotherapy resistance. Objective:To identify potential drug targets using comparative genomic microarray analysis of specimens in patients with PBL. Methods:PBL data from three different research groups were used; i.e. COG ANHL01P1 by our group (n=11), NCBI's GEO GSE10172 and GSE4475 (Klapper et al) (n=16), and GSE4732 (Dave/Staudt et al) (n=15). For comparison at equal weighted level, ANHL01P1 samples were validated by building a prediction model with Support Vector Machines using Klapper PBL as training database. RNAs were subjected to microarray studies (Affymetrix U133A_2) and analyzed by Agilent GeneSpring or Partek. Functional interpretation of the identified PBL genes was analyzed by Ingenuity Pathways Analysis. One-way ANOVA followed by Tukey test was used. To test our hypothesis, we knocked down DLEU1 and investigated cytoxan (CY) and rituximab effect on apoptotic rate of Ramos. Ramos BL cell lines were transiently transfected (24 hrs) with DLEU1 siRNA (5′-AUACUUGGCAUGAAUGAACUUAUGU-3′ and 3′-UAUGAACCGUACUUACUUGAAUACA-5′) (Day/Cairo SIOP 2008). The siRNA transfected cells were then treated with CY (0, 89.5, 895, 8950 nM) or rituximab (0, 4, 40, 400 mg/mL) for additional 4 hrs. Cells were evaluated for percent apoptosis using Annexin V-FITC and Propidium Iodide followed by FACS using BD LSRII. Results:1565 genes were identified (p<0.05), among which 376 genes showed no significant difference among three groups. High expression of cMYC (27F) and DLEU1 (9.9F) were detected. Pathway analysis indicates that these genes areinvolved in Toll-like receptor signaling (16 genes; p<0.01), including IRAK1 (22.9F), IFNAR2 (5.4F), NFKBIA (15.3F), and STAT1 (9.2F); JAK-STAT signaling (16 genes; p<0.01), including PTPN11 (25.6F), PTPN6 (8.8F), PIM1 (6.4F), and IL21R (2.9F); and MAPK signaling (19 genes; p<0.01) including MAP2K1 (11.8F), MAP3K7 (6.3F), MAPK9 (6.9F), and RAF1 (9.2F). There was a significant reduction in apoptosis in the CY-treated BL transfected DLEU1 siRNA vs mock control cells (89.5 nM CY: 10.26+0.23% reduction, p<0.05 to negative control; 895 nM CY, 10.86+0.67% reduction, p<0.01; 8950 nM, 9.85+0.32% reduction, p<0.05. There was a similar significant reduction in rituximab-induced apoptosis in the BL transfected DLEU1 siRNA vs mock control cells (4 mg/mL Rituximab: 25.45+2.55% reduction, p<0.01 to negative control; 40 mg/mL Rituximab, 18.31+5.13% reduction, p<0.04; 400 mg/mL rituximab, 32.33+1.77% reduction, p<0.02). Conclusions:Our pathway analysis of the 376 BL signature indicates that Toll-like receptor, JAK-STAT, and MAPK signaling pathways play important roles in gene regulation, anti-apoptosis, and proliferation, respectively, in PBL. The siRNA knock down of DLEU1 resulted in significantly less apoptosis on Ramos with CY or rituximab treatment. These data suggest that DLEU1 may in part play an important role in regulation of programmed cell death in BL and suggest a mechanism whereby BL with a 13q- abnormality may have a more aggressive clinical course. As CY or rituximab does not induce apoptosis in DLEU1 knockdown BL, the absence of DLEU1 may, in part, lead to drug resistance. Disclosures:No relevant conflicts of interest to declare.

Primary Bone Diffuse Large B-cell Lymphoma

Primary bone diffuse large B-cell lymphomas (PB-DLBCL) are uncommon extranodal lymphomas. Herein, we report the clinical, pathologic, immunohistochemical, and molecular features of 21 cases of PB-DLBCL. The mean age of the patients was 54 years (range: 13 to 85 y). The male and female ratio was 1.6:1. The tumors consisted of diffuse sheets of large atypical cells or a polymorphous mixture of small-to-large cells with large multilobated nuclei, fine chromatin, and inconspicuous to prominent nucleoli. Twelve (57%) cases were non-germinal center B (GCB) and 9 (43%) were GCB subtype based on immunohistochemical classification. B-cell lymphomas (BCL)-2 was positive in 17/21 (81%), TP53 in 11/21 (52%) positive and the mean MIB-1 index was 57%. Polymerase chain reaction showed 10 cases with immunoglobulin heavy-chain (IGH) and 4 cases with IGH/BCL-2 gene rearrangement. The fluorescence in-situ hybridization analyses showed 14% of cases with BCL-6, 19% of cases with BCL-2, and 9% of cases with C-MYC gene rearrangement. Age <60 years and complete response to initial treatment were significant predictors of survival outcome (P< or =0.05). Even though no association was observed between the subtype of PB-DLBCL (GCB vs. non-GCB), BCL2, TP53, MIB1 index and overall survival (P>0.05), due to small sample size, and variability in treatment received, this analysis may be interpreted with caution.

Complete remission of gastric Burkitt’s lymphoma aftereradication of Helicobacter pylori

Burkitt's lymphoma is a highly aggressive non-Hodgkin lymphoma, often presenting in extra-nodal sites. It generally has a poor spontaneous outcome and needs aggressive treatment with systemic and intrathecal chemotherapy. Occurrence at the gastric site is rare. We report the case of a 39-year old woman who presented with a prominent ulcerated lesion of the antrum corresponding histologically to a Burkitt's lymphoma associated with Helicobacter pylori (H pylori) infection. Interphase fluorescence in situ hybridization (FISH) demonstrated c-MYC gene rearrangement in tumour cells without BCL2 or BCL6 gene translocations. Ulcer healing and tumour regression with a complete histological response were obtained 8 wk after H pylori eradication. In spite of this complete remission, taking into account the high risk of recurrence, the patient received systemic and intrathecal chemotherapy. Two years later, the patient remained in complete remission. This is the first report of a gastric Burkitt's lymphoma responding to H pylori eradication. These findings raise the question of the potential role of H pylori in the pathogenesis of some gastric Burkitt's lymphomas, and show the importance of searching for and eradicating the bacteria in combination with conventional chemotherapy regimens.

A combination therapy with fludarabine, mitoxantrone and rituximab induces complete immunophenotypical remission in B-cell prolymphocytic leukaemia

Dear Editor, We present here the cases of two patients with B-cell prolymphocytic leukaemia (B-PLL), both expressing a translocation t(8;14)(q24;q32) with rearrangement of cmyc gene found in FISH analysis. A treatment protocol that includes six cycles of Fludarabine and Mitoxantrone followed by four injections of Rituximab allows obtaining complete cytological and immunophenotypical remission. B-PLL is a rare disease that accounts for less than 2% of chronic lymphatic leukaemia (CLL). The characteristics of the disease include high lymphocytosis, over 4 Gigas/L, with at least 55% cells described as prolymphocytes and splenomegaly. The prolymphocytes are medium-sized cells with prominent, round or oval nuclei and a single, central nucleolus [1]. Bone marrow aspirate shows a heavy infiltration by prolymphocytes with a variable degree of residual haematopoiesis. Immunophenotypically, there is a strong expression of the B-cell antigens CD19, CD20, CD21, CD79b and FMC7 and of surface IgM, whereas CD23 is negative and CD5 is variably expressed [1]. In cytogenetic studies, the implication of the 14q32 region of the chromosome 14 is found in 50% of the cases [1]. Translocation (11;14) is present in 25% of the cases, in particular t(11;14)(q13;q32) with a rearrangement of bcl1 protein. Other partners for the IgH gene of the 14q32 region of chromosome 14 could be genes from the regions 11q23 and 13q14 [1]. A translocation (8;14)(q24;q32) typically exists in type 3, B-cell acute lymphoblastic leukaemia and Burkitt lymphomas. This anomaly, which also involves cmyc, was found in only a few cases [2, 3]. B-PLL is not treated when asymptomatic or there are no signs of bone marrow deficiency. Treatment options are various, including combination chemotherapies with alkylating agents or purine nucleoside analogues as monotherapy. Monoclonal antibodies such as Rituximab and Alemtuzumab are also used. A 63-year-old man was first seen in October 2006 with symptoms of asthenia and dyspnoea. A physical examination revealed a splenomegaly grade 3 and lymphadenopathy. The blood count showed a significant anaemia (Hb 5,4g/dL), thrombocytopenia (108 Gigas/L) and a hyperleukocytosis at 247Gigas/L with 92% abnormal circulating lymphocytes, which presented the typical morphology and phenotype of B-PLL. Genetic studies revealed a complex caryotype (Fig. 1), involving a translocation t(8;14)(q24;q32) with a rearrangement of c-myc, confirmed by fluorescence in situ hybridisation analysis (Fig. 2). Ann Hematol (2009) 88:85–88 DOI 10.1007/s00277-008-0541-5

Increased expression of the c-myc gene may be related to the aggressive transformation of human myeloma cells

Alteration and abnormal expression of the c-myc oncogene were investigated in human multiple myeloma. Human myeloma cells were highly purified (more than 95%) from bone marrow aspirates in 14 cases of advanced multiple myelomas and one case of plasma cell leukaemia. Southern blotting revealed that a rearranged configuration of c-myc gene was found in only one case of them, but this was a novel truncation of the gene in its coding exon II; a rearranged 3.4 kb band was detected by digestion with Xba I using c-myc exon II probe, but no rearranged band was found using exon III probe. In this case, the truncated c-myc allele was not transcribed; normal sized (2.4 kb) c-myc mRNA was markedly expressed, but no aberrant mRNA was detected. On the other hand, by Northern blotting, the nine cases, including the case with the rearranged c-myc gene, showed increased expression of normal sized (2.4 kb) c-myc mRNA. Elevated c-myc mRNA expressions were well related to the in vitro proliferation (3H-TdR uptake), but not to IL-6 response. Interestingly, extremely high expressions of c-myc mRNA were detected in two cases of aggressive myelomas, including the case with the rearranged c-myc gene, and in one of plasma cell leukaemia. These two cases of aggressive myelomas were the ones who showed the markedly high 3H-TdR uptakes, and had the common clinical features with the formation of an extramedullary mass and very short survival. These results suggest that the activation of c-myc gene could induce high proliferative activities and the subsequent aggressive transformation of myeloma cells.

Plasmablastic Lymphoma of Head and Neck: Report of Two New Cases and Correlation with c-myc and IgVH Gene Mutation Status

Plasmablastic lymphoma (PBL) is a rare acquired immunodeficiency syndrome-associated non-Hodgkin's lymphoma (AIDS-NHL), with predilection for the mucosa of oral cavity. It usually has a plasmablastic morphology, expressing plasma cell-associated antigens with weak or absent expression of B-cell-associated markers. To further define the immunophenotypic and molecular genetics of these tumors, we investigated two cases of plasmablastic lymphomas of the head and neck for c-myc gene rearrangement and immunoglobulin heavy chain (IgV(H)) hypermutation status. For the first time we report a case of AIDS-related PBL that, by fluorescence in situ hybridization (FISH), shows a c-myc gene rearrangement. Although current literature suggests that most cases of c-myc gene rearranged AIDS-NHL are Burkitt's lymphoma, our case has an immunophenotype characteristic for PBL. In this case, IgV(H) hypermutation analysis showed a somatic hypermutation, indicative of germinal center transit. The concurrent B-cell immunophenotype of BCL-6(-)/CD138(+)/MUM-1(+) also suggests a post-germinal center B-cell origin of this lymphoma. The immunophenotype of our second case (BCL-6(-)/CD138(+)/MUM-1(+)) also suggests a post-germinal center B-cell origin. However, IgV(H) hypermutation analysis was not possible in this case.

Pediatric ALK+ anaplastic large cell lymphoma with t(3;8)(q26.2;q24) translocation and c‐myc rearrangement terminating in a leukemic phase

Pediatric ALK-positive anaplastic large cell lymphoma (ALK+ ALCL) is usually associated with a favorable prognosis. ALK+ ALCL associated with a leukemic phase is uncommon, but has been associated with an aggressive clinical course and unfavorable prognosis. Overexpression of c-myc has been shown to be a consistent finding in ALK+, but not ALK-negative ALCL (ALK- ALCL), and the c-myc gene is considered a downstream target of deregulated ALK signaling. We describe a pediatric ALK+ ALCL with a leukemic phase at relapse. Similar to other rare cases described in the literature, it followed an aggressive clinical course despite multiple regimens of chemotherapy and bone marrow transplantation. Lymphoma cells showed aberrant ALK expression and c-myc overexpression. In addition to the characteristic t(2;5)(p23;q35) translocation, a t(3;8)(q26.2;q24) translocation was also present, and c-myc gene rearrangement was confirmed by FISH analysis. The findings in this case demonstrate the association of peripheral blood leukemic involvement and aggressive clinical course, and suggest that other factors, such as c-myc rearrangement, may be responsible for the aggressive clinical behavior in ALK+ ALCL.

Ovarian Burkitt's lymphoma diagnosed by a combination of clinical features, morphology, immunophenotype, and molecular findings and successfully managed with surgery and chemotherapy

Ovarian involvement as an initial manifestation of lymphoma, without detectable extraovarian disease, is a rare occurrence. The diagnosis of ovarian lymphoma is almost invariably unsuspected until the tumor has been examined histologically. A 25-year-old null gravid woman presented with abdominal distension. Presence of abnormal lymphoid cells in pleural effusion led to presurgical assumption that the pelvic mass noted on computerized tomography examination might be an ovarian lymphoma. We performed left salpingo-oophorectomy. Clinical, histologic, and molecular examination revealed Burkitt's lymphoma of the ovary with c-myc gene rearrangement and mRNA expression of multiple cytokines. She received dose-intensified combination chemotherapy. She is alive and free of disease 30 months after the diagnosis. Immunophenotype and molecular findings allowed reliable discrimination of Burkitt's lymphoma from diffuse large B-cell lymphoma and other lymphomas. If an ovarian tumor is solid and suspected to be of lymphoid origin, we suggest that it is necessary to obtain samples for genetic examination at surgery. This strategy often provides important information to establish therapeutic regimen and predict patient prognosis.

Ovarian Burkitt's lymphoma diagnosed by a combination of clinical features, morphology, immunophenotype, and molecular findings and successfully managed with surgery and chemotherapy

Ovarian involvement as an initial manifestation of lymphoma, without detectable extraovarian disease, is a rare occurrence. The diagnosis of ovarian lymphoma is almost invariably unsuspected until the tumor has been examined histologically. A 25-year-old null gravid woman presented with abdominal distension. Presence of abnormal lymphoid cells in pleural effusion led to presurgical assumption that the pelvic mass noted on computerized tomography examination might be an ovarian lymphoma. We performed left salpingo-oophorectomy. Clinical, histologic, and molecular examination revealed Burkitt's lymphoma of the ovary with c-myc gene rearrangement and mRNA expression of multiple cytokines. She received dose-intensified combination chemotherapy. She is alive and free of disease 30 months after the diagnosis. Immunophenotype and molecular findings allowed reliable discrimination of Burkitt's lymphoma from diffuse large B-cell lymphoma and other lymphomas. If an ovarian tumor is solid and suspected to be of lymphoid origin, we suggest that it is necessary to obtain samples for genetic examination at surgery. This strategy often provides important information to establish therapeutic regimen and predict patient prognosis.

C-myc Abnormalities in Non-Hodgkins Lymphoma - A Retrospective Review of Cytogenetic Patterns and Clinical Impact of c-myc Abnormalities by FISH Testing.

Recent studies have shown that patients with aggressive histology non-hodgkins lymphoma (NHL) bearing translocations leading to aberrant expression of c-myc have a rapid clinical course and poor outcome, especially when accompanied by translocations involving BCL-2 such as t(14;18). We retrospectively reviewed patients with NHL in which abnormalities of the c-myc gene (rearrangement, extra copies, deletion) were found and correlated the cytogenetic abnormality with presenting characteristics and outcome.Methods: From May 2002 to June 2005, 138 samples were assessed for c-myc; 27 cases of NHL (excluding Burkitt [BL]and BL-like) with c-myc abnormalities were identified and included (25 tissue biopsies, 1 bone marrow, 1 CSF sample). Testing for c-myc abnormalities was initiated for: differential diagnosis of atypical BL or one or more of the following: high MIB-1 index (>70%) − 63%; aggressive morphological features − 81%; aggressive clinical course − 52%; suspected transformation of indolent lymphoma − 19%.Results: There were 12 males and 15 females included with an age range of 26–83 years; 67% had stage 3 or 4 disease and 38% had B-symptoms. The pathological diagnosis was follicular lymphoma (FL) in 3 cases, diffuse large B-cell lymphoma (DLBCL) in 22 cases [13 de novo, 5 transformed from FL, 4 recurrent/refractory] and 2 cases with both FL and DLBCL. FISH analysis for c-myc by the Vysis dual colour break-apart probe (8q24.1) revealed 12 cases with only extra copies (polyploidy) [group 1]; 13 with rearrangement (4 also had extra-copies) [group 2]; one with gene deletion and one with gene deletion and extra-copies. BCL-2 rearrangements were also detected in 22 cases (in group 1, 44%; group 2 69%). All patients were to receive multiagent anthracycline based chemotherapy; chemotherapy was initiated in 89% and one died before this could be given. Thus far, 41% of the patients have died and 19% are alive with refractory disease. The clinical course for group 1 after c-myc abnormality identified was as follows: refractory to chemotherapy - 6 cases, 5 in complete remission (CR), one spontaneous remission without treatment (FL). The median survival for group 1 was 5 months (range 1–21 months) and deaths occurred shortly after current diagnosis (range 1–7 months). The clinical course for group 2 was as follows: refractory to chemotherapy - 11 cases and 3 are currently in a CR. The median survival for group 2 was 9 months (follow-up 1–13 months) and again deaths occurred shortly after current diagnosis (range 1–9 months).Conclusions: Both c-myc gene rearrangement and extra copies of c-myc seem to confer poor prognosis in NHL. Not all patients with c-myc abnormalities have morphological features suggestive of Burkitt-like lymphoma, while some patients with only extra copies of c-myc did have Burkitt-like lymphoma features. Testing for c-myc abnormalities may be indicated not only in patients with aggressive histology but also in those with clinically aggressive patterns of disease, and prospective evaluation of a c-myc testing algorithm to evaluate impact on therapy is required.

Frequent aberrations of chromosome 8 in aggressive B-cell non-Hodgkin lymphoma

Translocations involving chromosome 8 are the most common aberrations in B-cell non-Hodgkin lymphoma (B-NHL). The presence of the typical t(8;14)(q24;q32) or its variants has been confirmed in all cases of Burkitt lymphoma (BL), in some cases of Burkitt-like lymphoma (BLL), and in diffuse large B-cell lymphoma (DLBCL). The alterations lead to deregulated expression of c-myc protein by a chromosomal translocation joining C-MYC gene with sequences from immunoglobulin (Ig) enhancers. The C-MYC gene rearrangement plays an essential role in leukemogenesis of BL and probably plays a part in other aggressive NHLs. The present study was undertaken to compare the cytogenetic features in cases of BL, BLL, and DLBCL. We detected chromosomal aberrations by G-banding and fluorescence in situ hybridization (FISH) painting in 10 cases of aggressive B-NHL and used FISH to visualize the C-MYC gene rearrangement. Chromosome 8 was most frequently involved in structural aberrations (8/10 cases), and 4 cases showed the typical t(8;14)(q24;q32). Only two of 5 patients suspected of having BL fulfilled all the criteria for this diagnosis; in the others, chromosome 8 was aberrant, but the absence of C-MYC rearrangement or the results of flow cytometry excluded the diagnosis of BL. All BLL cases showed C-MYC overexpression, but only one had a rearrangement of the C-MYC gene; the remaining cases showed other aberrations of chromosome 8. This study indicates that the mechanisms of C-MYC activation involved in BLL can be different from that for the BL.

Primary effusion lymphoma of the pericardial cavity carrying t(1;22)(q21;q11) and t(14;17)(q32;q23)

We report a case of primary effusion lymphoma (PEL) in a 75-year-old woman without human immunodeficiency virus or hepatitis C virus, which presented as fever, chest pain, and pericardial effusion. The lymphoma cells were positive for CD20 and CD79a, and were negative for CD3 and CD10. Genomic human herpes virus 8 (HHV-8) and Epstein-Barr virus were not detected in the lymphoma cells. Cytogenetic analysis showed complex abnormalities by the G-banding technique, and spectral karyotyping (SKY) analysis provided more detailed characterization of the chromosomal aberrations, including t(1;22)(q21;q11) and t(14;17)(q32;q23). We did not detect C-MYC gene rearrangement or BCL-2 expression. She was treated successfully with six courses of the CHOP regimen. The present case demonstrated a rare category of PEL that is not associated with HHV-8 or C-MYC gene rearrangement. In addition, SKY analysis disclosed cryptic chromosomal abnormalities involving 1q21 and 17q23.

Burkitt Transformation of Mantle Cell Lymphoma

The associated poor prognosis and potentially aggressive behavior of mantle cell lymphoma and its blastoid variants make differentiation from other non-Hodgkin B-cell lymphomas especially important. We present a case of mantle cell lymphoma with a marked leukemic component, which demonstrated both a typical nodular mantle cell pattern and Burkitt lymphoma within a single lymph node removed at the time of splenectomy. The presence of CD5, CD10, and Bcl-1 co-expression by immunohistochemistry and detectable t(11;14) and cMYC gene rearrangement by FISH analyses in the Burkitt region support a transformation of mantle cell lymphoma over a concomitant malignancy. A limited number of mantle cell lymphomas demonstrating dual t(11;14) and chromosome 8q24 cMYC gene rearrangements have been previously reported in the literature. They demonstrate an extremely aggressive course with a very poor prognosis. Although the accelerated terminal phase of this patient's clinical course mirrors these previous published cases; none have described the combined morphologic and immunophenotypic features of Burkitt lymphoma reported here. This case provides further support for the aggressive nature of these lymphomas and demonstrates the utility of flow cytometry, immunohistochemistry, and cytogenetic techniques in avoiding potential errors in their diagnosis, prognosis, and treatment.