Simonetta Kerim

Chronic myeloid leukemia: a prospective comparison of interphase fluorescence in situ hybridization and chromosome banding analysis for the definition of complete cytogenetic response, a study of the GIMEMA CML WP

In chronic myeloid leukemia, different methods are available to monitor the response to therapy: chromosome banding analysis (CBA), interphase fluorescence in situ hybridization (I-FISH), and real-time quantitative polymerase chain reaction (RT-Q-PCR). The GIMEMA CML WP (Gruppo Italiano Malattie Ematologiche Adulto Chronic Myeloid Leukemia Working Party) has performed a prospective study to compare CBA and I-FISH for the definition of complete cytogenetic response (CCgR). Samples (n = 664) were evaluated simultaneously by CBA and I-FISH. Of 537 cases in CCgR, the number of positive nuclei by I-FISH was less than 1% in 444 cases (82.7%). Of 451 cases with less than 1% positive nuclei by I-FISH, 444 (98.4%) were classified as CCgR by CBA. The major molecular response rate was significantly greater in cases with I-FISH less than 1% than in those with I-FISH 1% to 5% (66.8% vs 51.6%, P < .001) and in cases with CCgR and I-FISH less than 1% than in cases with CCgR and I-FISH 1% to 5% (66.1% vs 49.4%, P = .004). I-FISH is more sensitive than CBA and can be used to monitor CCgR. With appropriate probes, the cutoff value of I-FISH may be established at 1%. These trials are registered at http://www.clinicaltrials.gov as NCT00514488 and NCT00510926.

Deletions of the Derivative Chromosome 9 Do Not Influence the Response and the Outcome of Chronic Myeloid Leukemia in Early Chronic Phase Treated With Imatinib Mesylate: GIMEMA CML Working Party Analysis

PURPOSE Deletions of the derivative chromosome 9 [der(9)] have been associated with a poor prognosis in chronic myeloid leukemia (CML) across different treatment modalities. In the imatinib era, the prognostic impact of der(9) deletions has been evaluated mainly in patients with late chronic-phase (CP) CML, giving partially conflicting results. Few data are available in the early CP setting. For this reason, in 2006, the European LeukemiaNet recommendations still considered der(9) deletions as a candidate adverse prognostic factor and required a careful monitoring of the patient. PATIENTS AND METHODS To investigate the prognostic value of der(9) deletions in early CP CML, we performed an analysis of three prospective imatinib trials of the Italian Group for Hematological Malignancies of the Adult (GIMEMA) CML Working Party. RESULTS A fluorescent in situ hybridization (FISH) analysis of bone marrow cells was performed at diagnosis; der(9) deletions were detected in 60 (12%) of 521 evaluable patients. At 60 months, the cumulative incidence of complete cytogenetic response and major molecular response-and the probability of event-free survival, failure-free survival, progression-free survival, and overall survival-in patients with and without deletions were not statistically different. CONCLUSION Our data strongly support the notion that, when investigated by FISH, der(9) deletions are not a poor prognostic factor in patients with early CP CML treated with imatinib.

Translocation t(6;9) occurring in acute myelofibrosis, myelodysplastic syndrome, and acute nonlymphocytic leukemia suggests multipotent stem cell involvement☆

The cytological and cytogenetic features of six patients with myeloid neoplasia and t(6;9)(p23;q34) including a case of acute myelofibrosis (AMF), a refractory anemia with excess of blasts (RAEB), and four cases of acute nonlymphocytic leukemia (ANLL) are described. Two patients in this series, both affected by ANLL type M2, presented an increase of bone marrow basophils, suggesting that this cytological-cytogenetic association is not absolute and that it may be more frequently observed in ANLL with maturation. All patients with de novo ANLL showed associated myelodysplastic features, and one patient presented a dysmyelopoietic syndrome, later evolving into ANLL. The presence of the t(6;9) in a range of myeloid neoplasias, with either concurrent myelodysplastic features or a preleukemic phase in cases of ANLL, provide evidence that this chromosome aberration may always involve a multipotent myeloid stem cell. Data on toxic exposure of the patients suggests that myeloproliferative disorders with the t(6;9) may frequently represent environmentally induced neoplasias.

cFLIP expression correlates with tumour progression and patient outcome in non-Hodgkin lymphomas of low grade of malignancy

The present study investigated whether the expression of cellular Fas‐associated death domain‐like interleukin‐1β‐converting enzyme (FLICE) inhibitory protein (cFLIP) conveys prognostic information in non‐Hodgkin lymphomas (NHLs). cFLIP expression was quantified by immunohistochemistry and immunofluorescence in biopsy specimens from 86 NHL patients for whom clinical information was available. NHL malignancy was graded as high/intermediate or low according to the World Health Organization Classification of Lymphoid Neoplasms. cFLIP was positive in 23 of 45 high‐/intermediate‐grade NHLs and in 25 of 41 low‐grade NHLs. Negative expression of cFLIP was associated with the presence of apoptotic cells in the tumour mass, regardless of the histotype and of the malignancy grade. In NHLs positive for cFLIP, 11 of 23 (48%) high‐/intermediate‐grade cases and 18 of 25 (72%) low‐grade cases showed a bad outcome. In NHLs negative for cFLIP, only four of 22 (18%) high‐/intermediate‐grade patients and 12 of 16 (75%) low‐grade patients achieved complete remission. All these correlations were statistically significant. The correlation of cFLIP expression with clinical outcome was independent of therapy, whether or not it included anti‐CD20 antibody (Rituximab). The present findings strongly indicate that cFLIP is a reliable predictor of tumour progression and clinical prognosis in NHLs of low grade of malignancy.

Co-expression of plexin-B1 and Met in human breast and ovary tumours enhances the risk of progression

Background: Plex-B1, the receptor of Sema4D, has been implicated in tumour growth, angiogenesis and metastasis. The binding of Sema4D to Plex-B1 can trigger the activation of Met tyrosine kinase, thereby promoting cell dissociation and invasive growth. We tested the hypothesis that the expression of Plex-B1, either alone or in association with Met, can be of predictive value for tumour progression. Methods: The expression and distribution of Plex-B1 and Met were investigated by immunohistochemistry and immunofluorescence in 50 human neoplasias originating in the breast and ovary, and correlated with clinical–pathological data at diagnosis. Results: Plex-B1 and Met were individually expressed in 14% and in 24% of the tumours, respectively. Plex-B1 and Met were co-expressed in 24/50 cases (48%), and in the majority of these (83%) Met was tyrosine phosphorylated. The expression of Plex-B1 or Met alone showed no significant correlation with tumour aggressiveness, whereas advanced stage tumours (III–IV) frequently showed Plex-B1–Met double-positive (9/13). Tumours co-expressing Plex-B1 and Met were characterised by worse grading and higher incidence of lymph node metastases. Out of 22 tumours with lymph node metastases, as many as 19 were Plex-B1 and Met double-positive (p=0.0008), and 17 expressed phosphorylated Met (p=0.002). Conclusions: Plex-B1 assumes a predictive value for unfavourable outcome when co-expressed with Met.

Human herpesvirus type 7 in Hodgkin's disease

Several lines of evidence have pointed to the involvement of a viral agent in the pathogenesis of Hodgkins disease (HD). Therefore we investigated the presence of human herpesvirus type 7 (HHV‐7) in 53 cases of HD by polymerase chain reaction (PCR), DNA in situ hybridization (ISH) and immunohistochemistry. HHV‐7 DNA was frequently detected (68% of the cases) in HD biopsies by PCR independently of the histological type, whereas only 32% (P < 0.05) of positive cases were found in 19 reactive lymph nodes. However, by applying the quantitative PCR technique, the majority of the samples showed a low level of viral load. Moreover, ISH for HHV‐7 DNA was positive in a low number of small T lymphocytes and consistently negative in Hodgkin and Reed‐Sternberg (HRS) cells, which appeared negative for HHV‐7 also at immunohistochemistry.  These results indicate that the high frequency of HHV‐7 infection in HD: (i) is probably non‐productive, (ii) mainly involves small lymphocytes belonging to the T‐lineage, and (iii) is probably due to the recruitment of non‐malignant reactive cells in HD tissue.

Rearrangements of Immunoglobulin and Tcr Genes in Lymphoid Blast Crisis of Ph+ Chronic Myeloid-leukemia

Clonal rearrangements of immunoglobulin heavy chain genes as well as both T cell receptor (TCR) δ and γ genes were found in four cases of blast crisis of Ph + chronic myeloid leukaemia with unequivocal B cell precursor (common) immunophenotype. In one case, the TCR β chain gene was also rearranged. Although the developmental sequence of TCR δ, γ and β rearrangements in T lymphocytes appeared to be respected, a full phenotypic effect, characteristic of T cell was not observed in these otherwise typical ‘common’ blast cells. Cytogenetic analysis ruled out the occurrence of TCR rearrangement due to structural chromosome changes. A high incidence of unexpected TCR gene rearrangements has been previously reported in the de novo‘common’ acute lymphoblastic leukaemias (ALL). Our cases of chronic myeloid leukaemia (CML) in lymphoid blast crisis show that genotypic similarities may exist between these two haematological entities.

Heterogeneous immunoglobulin gene rearrangement in a B-chronic lymphocytic leukemia progressing into non-Hodgkin lymphoma (Richter syndrome).

Background. The relationship between chronic lymphocytic leukemia (CLL) and supervening non‐Hodgkin lymphoma is debated, as is whether a particular genomic pattern is related to the emergence of the terminal lymphoma. To investigate these features, the molecular organization of the immunoglobulin (Ig) gene region in a case during both the B‐CLL and Richter transformation phase was studied.

Trisomy 8 and an unbalanced t(5;17)(q11;p11) characterize two karyotypically independent clones in a case of idiopathic myelofibrosis evolving to acute nonlymphoid leukemia

In a patient with idiopathic myelofibrosis (MFI) that had progressed to acute nonlymphoid leukemia (ANLL) after a long-lasting cytotoxic treatment, we observed two karyotypically independent cell populations, one showing trisomy of chromosome 8 as the only anomaly and one with an unbalanced translocation t(5;17)(q11) resulting in partial monosomy of 5q and 17p. The overall karyotypic configuration suggested that chromosome changes occurred as secondary events during the multistep process of leukemogenesis. The probable sequence of cytogenetic events in this patient and a review of the literature indicated that the t(5;17) may represent a therapy-induced abnormality nonrandomly related to the terminal phase of myeloid disorders.

Translocation t(11;21)(q24;q11.2) is a new nonrandomly occurring chromosome change in myelodysplastic syndromes

An identical translocation, t(11;21)(q24;q11.2), has been observed in three patients with a myelodysplastic syndrome. In all cases, duplication of the 11q+ marker and loss of the normal chromosome 11 were observed either at diagnosis or during the evolution of the disease. This apparently characteristic chromosome abnormality has not been previously described.