Renata Woroniecka

CCND2 rearrangements are the most frequent genetic events in cyclin D1(-) mantle cell lymphoma.

Cyclin D1(-) mantle cell lymphomas (MCLs) are not well characterized, in part because of the difficulties in their recognition. SOX11 has been identified recently as a reliable biomarker of MCL that is also expressed in the cyclin D1(-) variant. We investigated 40 lymphomas with MCL morphology and immunophenotype that were negative for cyclin D1 expression/t(11;14)(q13;q32) but positive for SOX11. These tumors presented clinically with generalized lymphadenopathy, advanced stage, and poor outcome (5-year overall survival, 48%). Chromosomal rearrangements of the CCND2 locus were detected in 55% of the cases, with an IG gene as partner in 18 of 22, in particular with light chains (10 IGK@ and 5 IGL@). No mutations in the phosphorylation motifs of CCND1, CCND2, or CCND3 were detected. The global genomic profile and the high complexity of the 32 cyclin D1(-) SOX11(+) MCL patients analyzed by copy number arrays were similar to the conventional cyclin D1/SOX11 MCL. 17p deletions and high Ki67 expression conferred a significantly worse outcome for the patients. This comprehensive characterization of a large series of cyclin D1(-) MCL patients indicates that these tumors are clinically and biologically similar to the conventional cyclin D1(+) MCL and provides a basis for the proper identification and clinical management of these patients.

Partial trisomy 11, dup(11)(q23q13), as a defect characterizing lymphomas with Burkitt pathomorphology without MYC gene rearrangement

Burkitt lymphoma (BL) is an aggressive non-Hodgkin lymphoma characterized by specific morphological and immunophenotypic features. The basic genetic feature of BL is the rearrangement of MYC gene, visible as t(8;14)(q24;q32) translocation or its variant. However, some lymphomas with characteristic BL morphology are nowadays diagnosed as B-cell lymphoma unclassifiable with features intermediate between DLBCL and BL (Inter-DLBCL/BL) for biological or clinical reasons. We present four lymphomas without the MYC rearrangement presented typical Burkitt morphology, FCM immunophenotype with some deviations when compared to a typical BL. The cases were finally diagnosed as Inter-DLBCL/BL. All of them presented a recurrent abnormality within the chromosome 11: dup(11)(q23q13). We suppose that the dup(11)(q23q13), in absence of the MYC gene rearrangement, is connected with borderline lymphomas with a morphology similar or identical to that of the Burkitt lymphoma. Identifying such an aberration may be helpful in the diagnostics of Inter-DLBCL/BL eventually forming a distinct subgroup of lymphomas.

Cytogenetic and Flow Cytometry Evaluation of Richter Syndrome Reveals MYC, CDKN2A, IGH Alterations With Loss of CD52, CD62L and Increase of CD71 Antigen Expression as the Most Frequent Recurrent Abnormalities

OBJECTIVES Richter syndrome (RS) is a transformation of chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL) into high-grade lymphoma. There are only limited data on flow cytometry (FCM) and cytogenetics in RS. METHODS In this study, FCM, classic cytogenetics (CC), and fluorescence in situ hybridization (FISH) were performed in eight RS cases. RESULTS Most cases of RS were characterized by a loss/decrease of CD52 and CD62L and increased CD71 expression. CC identified complex karyotypes, with losses of 9/9p and 17/17p as the most frequent in four of seven cases. Seven RS cases demonstrated MYC abnormalities. Disruptions of CDKN2A and IGH were identified in five of seven and four of seven RS cases, respectively. CONCLUSIONS Newly diagnosed RS is an oncologic emergency, and a quick diagnostic decision is crucial in clinical practice. Therefore, in patients with CLL/SLL and rapidly enlarging asymmetric lymphadenopathy and/or extranodal tumors, we strongly advise FCM of fine-needle aspiration biopsy (FNAB) material, including CD62L, CD52, and CD71 analysis as well as assessment of karyotype and at least MYC abnormalities by FISH of the same FNAB material. Loss of CD52 expression in RS most likely predicts resistance to alemtuzumab therapy, which is frequently used in CLL.

Different prognosis of acute myeloid leukemia harboring monosomal karyotype with total or partial monosomies determined by FISH: Retrospective PALG study

A monosomal karyotype (MK) was identified by banding techniques (BT) in acute myeloid leukemia (AML). However, BT may be insufficient to determine the actual loss of a complete chromosome, especially in complex karyotypes. We have investigated the effect of monosomy type, total (MK-t) and partial (MK-p), reevaluated by FISH, on prognosis. We have found that complete remission rate and probability of overall survival at 1 year was higher in MK-p (n=27) than MK-t (n=15) group (40% vs. 15.4%, P=0.19 and 30% vs. 9%, P=0.046, respectively). Our results indicate for the first time that monosomy type influences the prognosis of MK-AML.

Karyotype changes during long-term targeted therapy of chronic myeloid leukemia with imatinib.

The main risk factors during imatinib therapy of chronic myeloid leukemia are still subject to discussion. A group of 39 patients was cytogenetically examined and monitored before and during long-term treatment with imatinib. The cytogenetic response was investigated using karyotype analysis and fluorescence in situ hybridisation method. Different therapy effects were shown for three subgroups distinguished before the start of treatment: patients with the sole translocation t(9;22) with a typical pattern of BCR/ABL fusion vs. patients with submicroscopic deletion in the fusion region ABL/BCR of the sole t(9;22) vs. patients with aberrations additional to t(9;22) and without submicroscopic deletion. Of the two group with sole t(9;22) the group with deletion in the ABL/BCL region suffered a poorer treatment outcome than the group without deletion. The risk of progression of cytogenetic changes in group with deletion was more than nine times higher than in patients with sole t(9;22) without deletion (statistically significant).

Unusual cyclin D1 positive marginal zone lymphoma of mediastinum.

We report a case of 43-yr-old Caucasian female with an unusual, cyclin D1 positive marginal zone lymphoma (MZL) of mucosa-associated lymphoid tissue (MALT) type of the mediastinum. To date, only about 30 cases of this entity have been published. They occur mainly in Asian females with a history of coexisung autoimmune disease. To our knowledge, this is the first case of medistinal MZL with cyclin D1 expression. In the span of 6 yr this patients tumor recurred three times, was surgically treated, and initially diagnosed as paraganglioma. The diagnosis was based on histopathological examination only. Our final diagnosis of MZL was made by combined evaluation of histopathology (HP), immunohistochemistry (IH), flow cytometry (FCM), fluorescence in situ hybridization (FISH), and molecular biology studies. We found a positive cyclin D1 reaction by IH and cyclin D1 mRNA (CCND1) overexpression by reverse transcription polymerase chain reaction (RT-PCR). Very high cyclin D1 to β-actin mRNA ratio in this case was comparable with the ratio, characteristic for mantle cell lymphoma (MCL). However, there was no translocation t(11;14) found by FISH and an immunophenotype by IH and FCM was consistent with MZL ruling out MCL diagnosis. In addition, our case differs from other, previously reported thymic MZL lymphoma cases by no autoimmune disease association, Caucasian origin, and the absence of the plasmacytic differentiation on both HP/IH.

Concomitance of monosomal karyotype with at least 5 chromosomal abnormalities is associated with dismal treatment outcome of AML patients with complex karyotype – retrospective analysis of Polish Adult Leukemia Group (PALG)

Abstract Monosomal karyotype (MK) and complex karyotype (CK) are poor prognostic factors in acute myeloid leukemia (AML). A comprehensive analysis of cytogenetic and clinical factors influencing an outcome of AML-CK+  was performed. The impact of cladribine containing induction on treatment results was also evaluated. We analyzed 125 patients with AML-CK+  treated within PALG protocols. MK was found in 75 (60%) individuals. The overall complete remission (CR) rate of 66 intensively treated patients was 62% vs. 28% in CK+ MK− and CK+ MK+  group (p = .01). No difference in CR rate was observed between DA and DAC arms. The overall survival (OS) in intensively treated patients was negatively influenced by MK, karyotype complexity (≥5 abnormalities), and WBC >20 G/L in multivariate analysis. The addition of cladribine to DA regimen improved OS only in MK− but not in MK+  group. In conclusion, concomitance of MK with ≥5 chromosomal abnormalities is associated with dismal treatment outcome in AMK-CK+.

Mantle cell lymphoma presenting with paraproteinemia.

Mantle cell lymphoma (MCL) is most frequently diagnosed by the routine histological (HP) and immunohistochemical (IH) examination. Herein we present a case of 47-yr-old female with general lymphadenopathy and leukemic blood picture. She was initially diagnosed with marginal zone lymphoma (MZL). This diagnosis was based not only on the HP/IH examination but also on the presence of IgM paraproteinemia. Flow cytometry (FCM) examination was helpful in making of the final diagnosis of MCL. Fluorescence in situ hybridization and cyclin D1 mRNA detection by RT-PCR additionally confirmed the MCL diagnosis. The IgM paraproteinemia is rare in MCL, although is a common feature of lymphoplasmacytoid lymphomas (LPL) and is considered a major characteristic of Waldenström’s macroglobulinemia (WM).

Acute panmyelosis with myelofibrosis with EVI1 amplification.

EVI1 is located on chromosome 3q26 and is up-regulated mostly through an inv(3)(q21q26) or t(3;3)(q21;q26). Chromosomal aberrations involving 3q26 comprise 1-2% of all acute myeloid leukemia (AML). These changes result in overexpression of the EVI1 oncogene. EVI1 transcriptional activation has been reported in up to 10% of AML patients, even in the absence of 3q26 changes, and is an independent indicator of adverse prognosis. Rearrangements of the EVI1 locus are often associated with monosomy 7. We present a case of acute panmyelosis with myelofibrosis with a unique EVI1 amplification within a derivative 8 chromosome, characterized by karyotyping and fluorescence in situ hybridization, conventional high resolution comparative genomic hybridization, as well as by gene expression studies. We conclude that EVI1 overexpression as a consequence of EVI1 gene amplification causes similar biological effects to the changes caused by the typical 3q26 aberrations such as an inv(3)(q21q26) or t(3;3)(q21;q26) with EVI1 gene rearrangements.

Comparison of cytogenetic changes between primary and relapsed patients with borderline tumors of the ovary

The aim of this work was to compare cytogenetic changes in primary and relapsed borderline tumors of the ovary. We analyzed 11 tumors (6 primary and 5 relapsed) by conventional GTG banding analysis and fluorescence in situ hybridization. The tumors studied were clinical stages I and III. Genomic imbalances were detected in both investigated groups. In the primary tumors group, only simple chromosome changes were detected. There were gains of chromosome 12, 7, and 8. The presence of additional copies of chromosomes 12 and 7 was independent of histologic subtype, whereas trisomy 8 appeared only in serous tumors. In the group of relapsed borderline tumors, besides trisomies 7 and 12, the structural aberrations of chromosomes 1, 6q, 7q, and 10q were revealed. Gains of tested oncogenes (CCND1 and MYC) have been demonstrated in both groups of investigated tumors. Gains of CCNC1 and MYC genes could be of prognostic value in borderline tumors, but this assumption requires further research.