Oncogene Amplification as an Incidental Finding in FISH Testing for Gene Rearrangements in Lymphoid Hematopoietic Neoplasms

Abstract Abstract 2505 Oncogene amplification resulting in overexpression, although common in solid tumors, is rare in hematopoietic neoplasms. This is particularly true in lymphoid neoplasms compared to AML where MYC, MLL or RUNX1 (AML1) amplification has been mostly seen, and to CML where BCR/ABL fusion gene amplification has been also reported. Typically, lymphoid neoplasms are tested at diagnosis by FISH for specific reciprocal chromosome translocations that lead to overexpression of deregulated oncogenes such as BCL1, BCL2, BCL6 and MYC in B-cell lymphoma and myeloma or BCR/ABL gene fusion in ALL. Nevertheless, we have unexpectedly seen oncogene amplification from time to time in our FISH lab. To study the incidence of gene amplification and its diagnostic and clinical implications, we retrospectively analyzed FISH results routinely performed on paraffin embedded lymphoma tissues, lymph nodes, bone marrow aspirates and peripheral bloods in the past three and half years using translocation probes for BCL2/IGH, BCL1/IGH, MYC/IGH and BCR/ABL and break apart probes for BCL6 and MYC. The highest amplification rate seen was in BCL2/IGH testing: 11 of the 1,710 cases were positive (0.643%). In 2 cases with follicular lymphoma (FL), BCL2 amplification presented as homogenously staining regions (hsr), one case with diffuse large B-cell lymphoma (DLBCL) showed double minutes (dmin), and two cases with FL had a pattern of combined hsr and dmin. Five cases with low grade FL intriguingly showed a similar pattern of BCL2/IGH translocation and concomitant amplification of the rearranged BCL2 as a small hsr. The remaining case diagnosed as Burkitt lymphoma (BL) was positive for MYC/IGH translocation and for BCL2 amplification present as large hsr. BCL6 amplification was observed in 4 of 1,537 cases tested (0.26%). In all these cases, amplification presented as hsr. One case diagnosed as EBV+ DLBLC was positive for MYC/IGH translocation and 3' BCL6 high level amplification. Another case with FL showed BCL6 rearrangement and 5' BCL6 amplification. BCL1 amplification with translocation was seen as hsr in 1 case with mantle cell lymphoma out of 2,898 tested. BCL1 amplification was observed in another case with plasma cell myeloma as hsr out of 3,413 tests performed. MYC gene amplification was positive in 3 of 2,186 (0.137%). One case with BL showed MYC rearrangement with a concomitant 3' deletion and 5' amplification. The second case diagnosed as B-cell lymphoma with features intermediate between DLBCL and BL showed highly amplified MYC gene as hsr. The third case was DLBCL with 15–50 copies of MYC gene per cell as dmin. Among 530 BCR/ABL tests ordered for acute leukemia or ALL, 2 (0.377%) cases with T cell lymphoblastic leukemia/lymphoma (T-ALL/LBL) showed ABL amplification as episomes (4–8 copies in one case; 6–30 in another). Both were abnormal by cytogenetics analysis, but negative for t(9;22)(q34;q11.2) and without dmin or hsr identified. Although detection rates of oncogene amplification seem to be very low with limited specific translocation probes applied to lymphoid neoplasms, they may well be higher when FISH signal patterns are analyzed more carefully. In our study, 5 of the 11 FL cases tested for IGH/BCL2 showed a peculiar pattern positive for the t(14;18)-IGH/BCL2 and a concomitant red signal amplification of BCL2, the size of which is usually small and may be overlooked under microscope. All these 5 cases had low grade FL, suggesting that oncogene amplification can be an early genomic event in lymphomagenesis. MYC gene amplification is generally considered a late stage genomic alteration. When MYC is rearranged, amplification of BCL2, BCL6 or BCL1 may need to be taken into consideration for disease stratification, or vice versa. For instance, so called “double-hit” or “triple-hit” lymphomas currently require two or three concurrent translocations for diagnosis. Oncogene amplification, equally important for deregulation/overexpression as a translocation, should be considered as a second or a third hit in diagnosis and differential diagnosis of B-cell lymphoma unclassifiable with features intermediate between DLBCL and BL. FISH is a very useful tool to identify episomal oncogene amplification. Episomes, unlike cytogenetically evident dmin and hrs, are invisible by chromosome analysis, but their detection is important as reported for integration of targeted tyrosine kinase inhibitors into chemotherapeutic regimens. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

A Study of the Expression of the LMP1 Oncoprotein in Low-Grade B Cell Lymphomas and Correlation with the Levels of Oxidative Stress

Blood ◽

10.1182/blood.v120.21.4833.4833 ◽

2012 ◽

Vol 120 (21) ◽

pp. 4833-4833

Author(s):

Panagiotis Theodorou Diamantopoulos ◽

Vasiliki Papadopoulou ◽

Aikaterini Polonyfi ◽

Athanasios G. Galanopoulos ◽

Fani Kalala ◽

...

Keyword(s):

Oxidative Stress ◽

B Cell ◽

Conflicts Of Interest ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Low Grade ◽

B Cell Lymphomas ◽

Ebv Infection ◽

Lmp1 Expression ◽

Apoptotic Pathways

Abstract Abstract 4833 Introduction. The Epstein-Barr virus has been implicated in the pathogenesis of certain human B-cell neoplasms, such as Burkitt's lymphoma, Hodgkin's disease and post-transplant lympho-proliferative disorders. Persistent latent EBV infection is, however, frequent and therefore its role is of interest in all types of B cell malignancies. In low-grade B cell lymphomas there are few reports for its potential role in higher grade transformation and its association with stereotypic BCRs in CLL. The mechanisms of EBV-associated B cell transformation are probably associated with its proteins expressed during latency; one of the most studied is the LMP1 oncoprotein, which is considered as an anti-apoptotic factor (activator of NF-êB). Recent studies, however, show evidence of coexisting apoptotic properties of LMP1. The level of oxidative stress reflects activation of caspase-mediated apoptotic pathways. Aims and methods. We measured the levels of oxidative stress in low-grade B cell lymphoma patient samples and correlated them with the expression of the LMP1 oncoprotein in order to study apoptotic functions of LMP1. Whole blood samples from 48 patients aged 51–87 (median age 74 years, 25 males, 23 females) without treatment in the previous six months were examined (chronic lymphocytic leukemia: 27, marginal zone lymphoma: 12, mantle cell lymphoma: 4, hairy cell leukemia: 2, follicular lymphoma: 2, lymphoplasmacytic lymphoma: 1). Latent EBV infection was detected with RT-PCR for the viral BXLF1 gene. LMP1 expression was quantitated with Real-Time PCR in EBV-positive patients. The levels of oxidative stress were quantitated in the sera of all patients with the use of a peroxide measuring kit (PerOx TOS/TOC kit by Immundiagnostik) and compared between the LMP1-positive (13) and LMP1-negative (35) group of patients with the use of 2-tailed Mann-Whitney test. Results. Of the fourty-eight (48) patients tested, nineteen (19) were EBV-positive. Thirteen (13) of the nineteen (19) EBV-positive ones expressed LMP1. Oxidative stress was found to be significantly higher in LMP1-negative vs LMP1-positive patients (372.3 vs 261.4 micromol/L, p=0.014). Discussion. The role of LMP1 expression is under investigation in the non EBV-related low grade B cell lymphomas. In the present study we examined a potential effect of LMP1 expression on oxidative stress and found that levels of oxidative stress were lower in LMP1-positive vs LMP1-negative patients with low-grade B cell lymphomas, reflecting an anti-apoptotic function of LMP1. In accordance with this result, LMP1 has been shown to upregulate BCL-2 using the NF-êB pathway. BCL-2 is a major inhibitor of the initiation of caspase-related apoptotic pathways and BCL-2 upregulation inhibits apoptosis resulting in lower levels of oxidative stress. However, in a study of sixty-four patients with low grade B cell lymphomas, we recently showed that LMP1 expression increases the levels of the apoptotic marker survivin, confirming that LMP1 may also possess an apoptotic function, as has been shown by another recent study on cell lines. Conclusion. The lower oxidative stress in the LMP1-expressing low grade B cell lymphoma samples shows evidence of an apoptotic function of the oncoprotein in this group of diseases. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

Autologous SCT In Patients (pts) with Chemotherapy-Resistant Diffuse Large B Cell Lymphoma (DLBCL) Followed by Maintenance with Rituximab.

Blood ◽

10.1182/blood.v116.21.4598.4598 ◽

2010 ◽

Vol 116 (21) ◽

pp. 4598-4598

Author(s):

Rodolfo Calixto ◽

Mauricio Ostronoff ◽

Fabiana Ostronoff ◽

Alexandre Sucupira ◽

Djenane Manso ◽

...

Keyword(s):

B Cell ◽

Median Time ◽

Conflicts Of Interest ◽

Cell Lymphoma ◽

Conditioning Regimen ◽

B Cell Lymphoma ◽

Low Grade ◽

Bulky Disease ◽

Pseudomonas Infection ◽

Neutrophil Engraftment

Abstract Abstract 4598 There are few data on the use of maintenance Rituximab after auto-SCT for patients with B cell lymphoma of aggressive histology that are resistant to chemotherapy. From May 2005 to March 2010, 25 consecutive pts underwent auto-SCT for DLBCL resistant to chemotherapy at our center. Median age 43 year (7 – 69 yrs); 11 patients were male. Two pts were HIV positive. Initial staging was II-B in 6 patients, II-EB in 4 patients, III-B 8 patients and IV-B in 7 patients. Initial IPI score was low-intermediate (13% of the pts), high-intermediate (58%) and high (29%). Nine of the 25 pts had bulky disease and 8 had visceral or bone marrow lymphomatous involvement. The median time from diagnosis to auto-SCT was 21 months (13 – 48 m). Prior to auto-SCT, 44% of the patients received 2 chemotherapy regimens and 56% received more than 2. All pts had chemotherapy-sensitive disease to the their last chemotherapy regimen prior to auto-SCT, with 80% and 20% of the pts achieving complete and partial remission prior to transplant, respectively. Sixteen out of 25 pts received Rituximab prior to auto-SCT; of those, 5 received Rituximab in the first line chemotherapy and 11 received it as part of the rescue chemotherapy regimens. The protocol was approved by our institutional review board and informed consent was obtained from each pt and or their guardians. Conditioning regimen consisted of Cyclophosphamide 1500 mg/m2 (D-6 to D-3), Etoposide 400 mg/m2 (D-6 to D-3) and Carmustin 150 mg/m2 (D-6 to D-4). The median CD34+ cells/kg infused was 2.9 × 106/Kg (1.9 - 8.5×106). All pts received G-CSF 10 micrograms/Kg/day SC from day +1 until neutrophil engraftment. Median time to neutrophil engraftment (ANC >500/mm3) was 8 days (5 – 17 d). Median time to platelet recover (>20,000/mm3) was 13 days (7 – 29 d). Transplant related mortality at day +100 was one in 25 pts (4%); this pt died due to multi-drug resistant Pseudomonas infection on day +17. Rituximab 375mg/m2 weekly for 4 weeks was administered as maintenance for a total of 4 cycles (16 doses); the cycles started on days +120, +240, +360 and +480 after auto-SCT. Twelve pts developed mild infusion reaction (tremor and rash). The hematological toxicity was low; grade II neutropenia occurred in 9 out of 25 pts. The neutropenic only occurred after the forth dose of the cycle with a median duration of 5 days (2 - 13). All pts received Bactrim and Acyclovir prophylaxis for one year after the auto-SCT. There were no viral infectious complications. Four of the 25 pts died (16%); one due to Pseudomonas infection; 3 due to relapsed disease which occurred at 6, 9 and 19 months after the transplant. Overall disease free survival was 75% with a median follow up of 31 months (6 - 55 mo). Of the five pts with refractory disease who had received Rituximab at some point prior to transplant, 2 pts relapsed and died due to refractory Lymphoma after the transplant, but 03 are alive in CR (9, 13, 21 months). Our data suggests that the administration of Rituximab as maintenance after auto-SCT for pts with DLBCL is well tolerated and it may decrease the incidence of relapse. Randomized studies are warranted to confirm the benefit of Rituximab as maintenance in the post auto-SCT setting to decrease relapse rate. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

Lymphoproliferative Disorders (LPD) in Individuals ≥80 Years of Age–Incidence, Response, and Survival

Blood ◽

10.1182/blood.v118.21.4945.4945 ◽

2011 ◽

Vol 118 (21) ◽

pp. 4945-4945

Author(s):

Fallon O France ◽

G. Chikkappa ◽

Donald Pasquale

Keyword(s):

B Cell ◽

Survival Data ◽

Conflicts Of Interest ◽

Cell Lymphoma ◽

Dose Intensity ◽

B Cell Lymphoma ◽

Low Grade ◽

Old Patients ◽

Large B Cell Lymphoma ◽

Large B Cell

Abstract Abstract 4945 Introduction Clinical trials describing LPD therapy and outcomes overwhelmingly exclude individuals ≥80 years of age. Thus, ability to deliver LPD therapy to this age group is not defined, and tolerance to therapy and response rates and survival data are not available. Methods We retrospectively identified all individuals diagnosed with any LPD at ≥80 years of age at our institution between 1997 and 2010. Data included age at diagnosis, diagnosis, therapy for LPD, comorbidities (disease count), survival from date of diagnosis of LPD, chemotherapy dose intensity (percent of 100% dose based on standard regimen dosing), hospitalization for any reason during chemotherapy, and response. Survival was analyzed by Kaplan-Meier analysis and data sets were compared using Student's t-test or Chi-square as appropriate. Results Of a total of 518 individuals diagnosed with any LPD, we identified 33(6.4%) who were diagnosed ≥80 years of age. All were male consistent with our veteran population. Data is illustrated in tables 1 and 2. Fifteen (15) had Large B-cell lymphoma, 4 CLL/SLL, 2 follicular, 2 mantle zone, 1 marginal zone, 2 lymphoplasmacytic, 3 T-cell. Two (2) could not be classified despite AFIP review. Twenty-two (22) individuals were treated with 73 cycles of chemotherapy (25 CHOP, 30 R-CHOP, 3 CVP, 11 R-CVP, 4 PO cyclophosphamide). One (1) individual with LBCL had no evidence of disease following excisional biopsy and declined further therapy, and 1 received radiotherapy. A total of 27 (82%) patients died during observation. Deaths were predominantly due to LPD. Of those not treated, 5 of 9 had low-grade LPD and were among longer-term survivors. Median survival, illustrated in the figure, was 18.8 months and was not different between treated and untreated group. This compares with 72.7 months predicted survival for an average 84-year old (WWW.SSA.GOV). Discussion LPD is common in individuals ≥80 years of age with large B-cell lymphoma being most common in our population. Despite multiple co-morbidities, the data show that reasonable dose intensity combination chemotherapy may be delivered to treat octogenarians, however hospitalization was required for 1/3 patients during chemotherapy. While survival was not different between treated and untreated groups, the individuals who were treated likely had more severe or advanced stage LPD. Prospective studies with sufficient number of individuals within each diagnostic category should be done to clarify benefits of chemotherapy in ≥80 year old patients with LPD. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

The Separate Diagnosis of Hodgkin Lymphoma (HL) and Non-Hodgkin Lymphoma (NHL) In a Single Patient May Not Signify a Common Clonal Origin

Blood ◽

10.1182/blood.v116.21.4824.4824 ◽

2010 ◽

Vol 116 (21) ◽

pp. 4824-4824

Author(s):

Chezi Ganzel ◽

Galina Pogrebijsky ◽

Svetlana Krichevsky ◽

Tzahi Neuman ◽

Dina Ben-Yehuda

Keyword(s):

B Cell ◽

Hodgkin Lymphoma ◽

Gene Rearrangement ◽

Conflicts Of Interest ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Low Grade ◽

Primary Disorder ◽

Clonal Origin ◽

The Mean

Abstract Abstract 4824 Background: HL and NHL have traditionally been considered as two distinct entities. However, there have been reported rare cases of patients that over time develop both diseases. It is an unresolved issue whether the origin of the two diseases is from the same clone. The literature is replete with anecdotal reports but it has never been prospectively or retrospectively evaluated in consecutive patients from a large series. In this study we attempted to retrospectively investigate this phenomenon by reviewing the clinical and molecular aspects of a group of patients who developed both lymphomas. Although not absolutely definitive, a differing gene rearrangement (GR) pattern is currently thought to be highly suggestive of a different clonal origin. Methods: Study patients were all patients treated at the Hadassah University Medical Center, Jerusalem, Israel, who developed both kinds of lymphoma throughout their lives and were treated for at least one of the lymphomas during the period 1989–2010. The clinical and pathologic records of these patients were reviewed. Archival, formalin fixed, paraffin embedded tissue samples from all the patients that had available samples from both diseases were obtained. The rearranged immunoglobulin heavy-chain variable region genes from both diagnoses were amplified by polymerase chain reaction (PCR) and were compared to each other. Results: There were 26 patients who presented with two diagnoses. Twelve had HL as the primary disorder and the majority of these (75%) presented with aggressive lymphoma as the second lymphoma. The mean survival from the second lymphoma was 4.06 years. Five patients are still alive. In contrast, in the 11 patients where NHL was the primary disorder, this was usually (82%) of low grade histology. The mean survival in this case was 2.16 years. Four patients are still alive. Three patients were diagnosed concurrently with both diseases. For 11 patients there were available diagnostic samples for molecular analyses from both diagnoses of HL and NHL. In 6 of these 11 patients gene rearrangement studies were informative providing data for both diagnoses. The same GR was not found in any of the 6 patients. DLBCL=Diffuse Large B Cell Lymphoma, CLL/SLL=Chronic Lymphocytic Leukemia/Small Lymphocytic Lymphoma, marginal=marginal zone lymphoma, low grade=low grade B cell lymphoma unspecified Although the numbers are small, these data suggest that it is likely that in the case of two different lymphoproliferative disorders they are of separate clonal origin. Conclusions: The development of HL and NHL at different time points should be assumed to be a different biologic disease, until proven otherwise. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

MYC gene amplification in double minute chromosomes in an aggressive large B-cell lymphoma with leukemic presentation: a case report

Cancer Genetics and Cytogenetics ◽

10.1016/j.cancergencyto.2009.04.002 ◽

2009 ◽

Vol 192 (2) ◽

pp. 76-78 ◽

Cited By ~ 3

Author(s):

F. Alseraye ◽

R. Padmore ◽

M. Wozniak ◽

J. McGowan-Jordan

Keyword(s):

Case Report ◽

B Cell ◽

Gene Amplification ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Large B Cell Lymphoma ◽

Double Minute ◽

Myc Gene ◽

Large B Cell ◽

Double Minute Chromosomes

Download Full-text

Burkitt Lymphoma Arising From Lymphoplasmacytic Lymphoma Following Acquisition of MYC Translocation and Loss of the ETV6 Tumor Suppressor Gene

Archives of Pathology & Laboratory Medicine ◽

10.5858/arpa.2011-0429-cr ◽

2013 ◽

Vol 137 (1) ◽

pp. 130-133 ◽

Cited By ~ 7

Author(s):

Deniz Peker ◽

Brian Quigley ◽

Dahui Qin ◽

Peter Papenhausen ◽

Ling Zhang

Keyword(s):

B Cell ◽

Burkitt Lymphoma ◽

Cell Lymphoma ◽

Lymphoblastic Leukemia ◽

Bone Marrow Involvement ◽

B Cell Lymphoma ◽

Suppressor Gene ◽

Neck Mass ◽

Low Grade ◽

Lymphoplasmacytic Lymphoma

Lymphoplasmacytic lymphoma is a mature B-cell lymphoma with variable plasmacytic differentiation that displays an indolent clinical course. Its transformation to a high-grade B-cell lymphoma may occur uncommonly. Although acquisition of a MYC translocation could result in transformation of a low-grade lymphoma into diffuse large B-cell lymphoma, Burkitt lymphoma, or B-lymphoblastic leukemia, to our knowledge the latter 2 transformations have not been well documented in lymphoplasmacytic lymphoma. We report the case of a 70-year-old woman with a 9-year history of lymphoplasmacytic lymphoma/Waldenström macroglobulinemia who presented with rapid enlargement of a left neck mass and pancytopenia, which was diagnosed as Burkitt lymphoma with extensive bone marrow involvement. A series of histopathologic, molecular, and cytogenetic evaluations proved a cytogenetic evolution including t(8;14)(q24;q32)/MYC-IgH and identical clonal B-cell gene rearrangements from the 2 distinct lymphomas, confirming stage 4 aggressive Burkitt lymphoma arising from lymphoplasmacytic lymphoma.

Download Full-text

Introduction to a Special Issue on Low-Grade B Cell Lymphoma in the Spleen

Current Oncology ◽

10.3390/curroncol29010011 ◽

2021 ◽

Vol 29 (1) ◽

pp. 130-131

Author(s):

Victor E. Nava

Keyword(s):

B Cell ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Lymphoproliferative Disorders ◽

Molecular Medicine ◽

Low Grade ◽

Special Issue ◽

Lymphoid Neoplasms ◽

Splenic Lymphoma

Malignant lymphoproliferative disorders in the spleen may be primary (usually designated as splenic lymphoma) or secondary (due to progression of nodal or extra nodal lymphoid neoplasms) and represent an underestimated cause of splenomegaly, partially due to the decreasing frequency of splenectomy in our era of personalized molecular medicine [...]

Download Full-text

Tumor staging in a Beagle dog with concomitant large B-cell lymphoma and T-cell acute lymphoblastic leukemia

Journal of Veterinary Diagnostic Investigation ◽

10.1177/10406387211011024 ◽

2021 ◽

pp. 104063872110110

Author(s):

Alessandro Ferrari ◽

Marzia Cozzi ◽

Luca Aresu ◽

Valeria Martini

Keyword(s):

Bone Marrow ◽

Lymph Node ◽

T Cell ◽

B Cell ◽

Cell Lymphoma ◽

Tumor Staging ◽

Lymphoblastic Leukemia ◽

B Cell Lymphoma ◽

Large B Cell Lymphoma ◽

Cell Acute Lymphoblastic Leukemia

An 8-y-old spayed female Beagle dog was presented with peripheral lymphadenomegaly. Lymph node cytology and flow cytometry led to the diagnosis of large B-cell lymphoma (LBCL). We detected minimal percentages of LBCL cells in peripheral blood and bone marrow samples. However, a monomorphic population of neoplastic cells different from those found in the lymph node was found in the bone marrow. T-cell acute lymphoblastic leukemia was suspected based on flow cytometric immunophenotyping. PCR for antigen receptor rearrangement (PARR) revealed clonal rearrangement of both B-cell and T-cell receptors, and the presence of both neoplastic clones in the lymph node, peripheral blood, and bone marrow. The dog was treated with multi-agent chemotherapy but died 46 d following diagnosis. Tumor staging and patient classification are needed to accurately establish a prognosis and select the most appropriate therapeutic protocol.

Download Full-text