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Michael J. Keating - One of the best experts on this subject based on the ideXlab platform.
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Trisomy 12 chronic lymphocytic leukemia expresses a unique set of activated and targetable pathways
Haematologica, 2018Co-Authors: Lynne V. Abruzzo, Michael J. Keating, Carmen D Herling, George A Calin, Christopher C Oakes, Lynn L Barron, Haley E Banks, Vikram Katju, Kevin R CoombesAbstract:Although Trisomy 12 (+12) chronic lymphocytic leukemia (CLL) comprises about 20% of cases, relatively little is known about its pathophysiology. These cases often demonstrate atypical morphological and immunophenotypic features, high proliferative rates, unmutated immunoglobulin heavy chain variable region genes, and a high frequency of NOTCH1 mutation. Patients with +12 CLL have an intermediate prognosis, and show higher incidences of thrombocytopenia, Richter transformation, and other secondary cancers. Despite these important differences, relatively few transcriptional profiling studies have focused on identifying dysregulated pathways that characterize +12 CLL, and most have used a hierarchical cytogenetic classification in which cases with more than one recurrent abnormality are categorized according to the abnormality with the poorest prognosis. In this study, we sought to identify protein-coding genes whose expression contributes to the unique pathophysiology of +12 CLL. To exclude the likely confounding effects of multiple cytogenetic abnormalities on gene expression, our +12 patient cohort had +12 as the sole abnormality. We profiled samples obtained from 147 treatment-naive patients. We compared cases with +12 as the only cytogenetic abnormality to cases with only del(13q), del(11q), or diploid cytogenetics using independent discovery (n=97) and validation (n=50) sets. We demonstrate that CLL cases with +12 as the sole abnormality express a unique set of activated pathways compared to other cytogenetic subtypes. Among these pathways, we identify the NFAT signaling pathway and the immune checkpoint molecule, NT5E (CD73), which may represent new therapeutic targets.
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Second Cancers and Richter Transformation Are the Leading Causes of Death in Patients With Trisomy 12 Chronic Lymphocytic Leukemia
Clinical Lymphoma Myeloma and Leukemia, 2015Co-Authors: Paolo Strati, Lynne V. Abruzzo, Alessandra Ferrajoli, William G. Wierda, Susan O'brien, Michael J. KeatingAbstract:Abstract Background Trisomy 12 (+12) is detected by fluorescence in-situ hybridization (FISH) analysis in up to 20% of patients with chronic lymphocytic leukemia (CLL). Patients with +12 are known to have unique features and to carry an intermediate prognosis. Patients and Methods In order to better define this large group, we reviewed the characteristics of 250 untreated patients with +12. Results When compared to 516 untreated patients negative for +12 by FISH, patients with +12 showed a higher incidence of thrombocytopenia, Richter transformation, and second malignant neoplasms (SMN), in addition to the expected increased rate of CD38 positivity and atypical immunophenotype. At a median follow-up of 51 months, 57% of patients needed first-line treatment; median time to first treatment was 38 months, and on multivariate analysis (MVA), it was found to be shorter in patients with advanced Rai stage, palpable splenomegaly, and deletion of 14q by conventional cytogenetic analysis. The overall response rate with first-line treatment was 94%. The median failure-free survival has not been reached, but on MVA, it was found to be shorter in patients whose disease responded in a manner other than complete remission or with FISH negativity for deletion 13q. The median overall survival for the entire group has not been reached, but MVA revealed it to be shorter in patients with an absolute lymphocyte count of > 30 × 10 9 /L or who developed SMN. Eighteen deaths have been observed so far, and Richter transformation and SMN were the leading causes of death (3 and 6, respectively). Conclusion Patients with +12 CLL show characteristic clinical and biologic features, and may benefit from increased surveillance for second cancers.
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Trisomy 12 clls progress through notch1 mutations
Leukemia, 2013Co-Authors: Veronica Balatti, Susan Lerner, Laura Z Rassenti, Arianna Bottoni, Alexey Palamarchuk, Hansjuerg Alder, Lara Rizzotto, Luciano Cascione, Michael J. KeatingAbstract:B-cell chronic lymphocytic leukemia (CLL) is the most common adult leukemia in Western societies.1 CLL cases can be subgrouped into two types, aggressive or indolent, defined as cases that express high levels of Zeta-chain-associated protein kinase 70 (ZAP70) and unmutated immunoglobulin heavy-chain variable-region genes (IGHV), or low to negligible ZAP70 and mutated IGHV. Chromosomal aberrations can be identified in more than 80% of patients.1 The most frequent genetic alterations include deletion/ inactivation of 13q14 (450%), deletion of 11q22–23 (18%), Trisomy of 12 (12–16%) and deletion 17p (7–10%).1 Two recent studies reported whole-genome sequencing of CLL samples and found NOTCH1 is the most frequently mutated gene in CLL,2,3 although we found a mutation frequency of only 3.5% in a large cohort of CLL.4 Our recent report showed that NOTCH1 mutations in CLL associate with Trisomy 12.4 We found 41.9% NOTCH1 mutation frequency in aggressive Trisomy 12 CLL cases, suggesting that activation of NOTCH1 has a critical role in IGVH unmutated/ ZAP70 + Trisomy 12 CLL.4 NOTCH1 encodes a class I transmembrane protein functioning as a ligand-activated transcription factor.5,6 Upon ligand binding, Notch1 undergoes several proteolytic cleavages resulting in translocation of the Notch1 intracellular domain (ICN) to the nucleus where it has an important role in cell differentiation, proliferation and apoptosis, leading to transcriptional activation of multiple target genes, including MYC.7 ICN contains PEST domain targeting ICN for ubiquitinylation and degradation.5,6 Almost all NOTCH1 mutations in CLL are represented by the two base deletion frame-shift, resulting in a truncated constantly active protein lacking the C-teminal PEST degradation domain.2,3 On the basis of our previous findings, we investigated whether NOTCH1 mutations are early events in Trisomy 12 CLL, or these mutations are associated with CLL progression. Mutations were most commonly observed in 100% of cells with one WT allele and one mutant allele, but we also found samples with different mutation frequencies.4 Looking for change in mutation frequencies, we sequenced serial samples of 78 Trisomy 12 CLL patients, and found 10 informative cases (Table 1). First, we studied two samples from each patient taken as far as possible. Patients were than subgrouped in two categories: patients with constant NOTCH1 mutation rate and patients with changes in NOTCH1 mutation rate. Only samples showing changes in NOTCH1 mutation rate were considered informative. Clinical data were obtained to verify the disease progression and middle time points of these patients were tested to verify changes in NOTCH1 mutation rate during progression. In these cases mutation frequency varied from 100% WT alleles (both alleles were WT) to 10% WT allele and 90% mutated allele (in most cells both alleles were mutated) (Supplementary Figure 1). Table 1 Detailed description of informative CLL samples showing that NOTCH1 mutations associate with CLL progression Four cases (28, 52, 56 and 66) did not show mutations at the first time points, however, increased mutation frequency was evident at later time points. In all these cases increases in NOTCH1 mutation frequencies were associated with progression of the disease. Patient 56 showed increase in the mutant allele from 5 to 20%, while white blood cell count (WBC) increased from 5.1 to 53×109/l, and beta(2)-microglobulin (B2M) increased from 2.7 to 3.8 mg/dl. Clinical data supported CLL progression between the time points provided. Patient was first diagnosed in 2001. In April 2009, patient was treated with flurabine and rituximab and showed 0% of NOTCH1 mutation rate. After successful treatment ended in September 2009, NOTCH1 mutation rate rise to 5% as observed in 2010. Patient relapsed and progressed with increase of NOTCH1 mutation to 20% in 2011. In August 2011, patient reinitiated fludarabine and rituximab treatment. Patient 66 showed increase in the mutant allele from 0 to 90% while WBC increased from 4.6 to 18.6 × 109/l. Patient was treated before the first time point achieving complete response and showed wild-type (WT) NOTCH1 in April 2009. However, contemporaneously with raising of percentage of NOTCH1 mutation rate to 90%, patient developed progressive disease. Bone marrow biopsy revealed presence of lymphocytes constituting 50–60% of overall cellularity, 39% of them having immunophenotype consistent with CLL at flow cytometry evaluation. Patient 52 showed increase in the mutant allele from 0 to 50%, while WBC increased from 4.8 to 13.2 × 109/l, and B2M increased from 2.5 to 7.3 mg/dl. Patient was initially treated in 2004 and achieved partial response. In 2005, NOTCH1 gene sequence was WT. In 2007, indolent relapse of disease with hemolytic anemia occurred and patient was treated with rituxan. In April 2008, patient relapsed again, disease progressed and NOTCH1 mutation rise to 50%. Consecutively, patient was treated with oxiplatin, fludarabine, cytarabine and rituximab. Five other cases (11, 15, 65, 69 and 77) also showed increased mutation frequency with disease progression, although in these cases mutations were already present in the first time point samples. Interestingly, in three patients (15, 65 and 36) decreased mutation frequency was associated with treatment response. Patient 15 showed decrease in the mutant allele from 35 to 0%, while WBC decreased after treatment with fludarabine, rituximab and cyclophosphamide from 439 to 3.5 × 109/l, and B2M decreased from 8.2 to 1.9mg/dl. Patient 36 showed decrease in the mutant allele from 50 to 0%, while WBC decreased after treatment with cyclophosphamide, fludarabine, alemtuzumab and rituximab from 56.8 to 3.4 × 109/l, and B2M decreased from 5.1 to 2.5mg/dl. To investigate the molecular consequences of the expression of constantly active Notch1 protein in Trisomy 12 CLLs, we performed genome-wide Affymetrix array analysis by comparing NOTCH1 WT and NOTCH1-mutated samples. To insure reliability of the results all chosen samples were IGVH unmutated/ZAP70 + , and >50% of cells showed Trisomy 12 (Supplementary Table 1). Ten of ten NOTCH1 WT samples and six of seven NOTCH1-mutated samples showed tight clustering (Supplementary Figure 2A), the remaining NOTCH1-mutated sample (Mut1) showed only partial clustering (Supplementary Figure 2A). Among 20 most upregulated genes in NOTCH1-mutated samples, we did not find any genes with known oncogenic function. However, 10 of the 20 most downregulated genes in NOTCH1-mutated samples have known tumor suppressor or pro-apoptotic function (Table 2). Among these downregulated genes, we found three members of the FOS gene family associated with apoptotic cell death in CLL8(c-FOS, FOSB and FOSL2), CDKN1A (p21), a p53-dependent cell cycle inhibitor,9 and NR4A3 (NOR1), a known inducer of apoptosis in lymphoid cells10 (Table 2). Microarray data were confirmed by real-time reverse-transcription PCR for the following genes: CDKN1A (p21), c-FOS and FOSB (Supplementary Figure 2B). Thus, we concluded that activated Notch1 inhibits multiple tumor suppressors in Trisomy 12 CLLs. Table 2 Ten out of 20 most downregulated genes in NOTCH1-mutated samples have known tumor suppressor or pro-apoptotic function. As recent studies identified NOTCH1 as the most mutated protein-coding gene in CLL,2,4 it is important to determine whether NOTCH1 mutations are essential in the initiation of CLL, or whether these mutations are associated with progression of Trisomy 12 CLLs. Here we found nine CLL cases showing increase of NOTCH1 mutation frequencies associated with severity of the disease including four cases showing WT NOTCH1 at initial time points. Activation of NOTCH1 appears to be involved in downregulation of tumor suppressor and apoptotic key factors, accelerating the progression of the disease. Taken together, these data indicate that NOTCH1 mutations do not cause CLL but rather associate with CLL progression leading to more aggressive form of the disease with poor outcome.
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biological and clinical features of patients with chronic lymphocytic leukemia bearing Trisomy 12
Blood, 2012Co-Authors: Paolo Strati, Susan Lerner, Lynne V. Abruzzo, Alessandra Ferrajoli, William G. Wierda, Susan Obrien, Hagop M Kantarjian, Michael J. KeatingAbstract:Abstract Abstract 3907 Cytogenetic abnormalities are among the most important predictors of clinical course and response to therapy in patients (pts) with chronic lymphocytic leukemia (CLL). Conventional chromosome banding (CBA) and fluorescence in situ hybridization (FISH) analyses detect abnormalities in 40–50% and 80% of pts, respectively. Trisomy 12 (+12), observed in ∼20% of CLL pts by FISH, is associated with atypical morphology and immunophenotype, and a more aggressive clinical course. We, therefore, review the clinical characteristics and outcome of 312 CLL pts with +12 evaluated at our center between 1988 and 2011. FISH analysis for common abnormalities associated with CLL was performed on interphase nuclei obtained from cultured bone marrow cells using a multi-color probe panel designed to detect deletions of 11q22.3 (ATM), 13q14.3 (D13S319), 13q34 (LAMP1), 17p13.1 (TP53) and Trisomy 12 (12p11.1-q11) (Abbott Molecular, Abbott Park, IL). Survival curves were calculated using Kaplan-Meier estimates and compared using the log-rank test. Differences were considered significant for p Patient characteristics at diagnosis are presented in Table 1 . Of 215 pts assessed by both CBA and FISH, 105 were positive for +12 by both analyses and 110 were positive only by FISH. By CBA (112 pts, including 7 assessed only by CBA), +12 was the sole abnormality in 52 pts (47%); +12 was associated with +19 in 17 pts (16%), with del(14q) in 9 pts (8%), with +18 in 8 pts (7%), with +8 in 3 pts (3%), with del(13q) in 3 pts (3%), with t(14;19)(q32;q13) in 3 pts (3%) and with other abnormalities in 17 pts (13%). By FISH (287 pts), +12 was the sole abnormality in 225 pts (78%) and was associated with del(13q) in 62 pts (22%). The median number of interphase nuclei positive for +12 by FISH was 47% (range, 5–93%). One-hundred-eighty-seven pts (60%) needed treatment, with a median Time-To-Treatment (TTT) of 46 months (range, 35–56). The TTT was significantly shorter in pts with Rai stage III-IV disease, splenomegaly, lymphadenopathy, B2m > 4 mg/L, CD38+, ZAP70+, +12 detected by both CBA and FISH, and +12 associated with del(14q) or t(14;19). All 187 pts with progressive disease received treatment: 105 with an FCR-based regimen, 28 with rituximab(R)-based therapy (R+ GM-CSF or R+ methylprednisolone), and 28 with investigational drugs (Lenalidomide, R+ lenalidomide, GS101, or Ibrutinib). Overall response rate was 98%, 89% and 96%, respectively, whereas complete remission rate was 87%, 11% and 36%, respectively. Fifty-five pts failed first-line treatment; their median Failure-Free Survival (FFS) was 27 months (range, 0–87). The FFS was significantly longer in pts who received FCR-based regimens (p Fig 1 ). The median overall survival (OS) has not been reached, and only 33 pts have died. The OS was significantly shorter in pts older than 65 years, with ALC > 30,000, and with a median +12 positivity in >30% of interphase nuclei by FISH. A trend toward longer OS was observed for pts with +12 associated with +19 (p=0.07). Richter’s Syndrome (RS) and second malignancies (SM) were the leading causes of death (5 and 13 of 33 deaths, respectively). RS was reported in 12 pts (4%), after a median time of 36 months from diagnosis. SM was reported in 31 pts (10%), after a median time of 30 months from diagnosis. At the time of diagnosis of SM, 13 patients had received a therapy for CLL and 18 were untreated. In conclusion, pts with CLL and +12 have unique laboratory and clinical features. A high proportion develops progressive disease and requires treatment. Among available therapies, FCR-based regimens are associated with a longer FFS. A high rate of SM is observed in pts with +12, including in pts who have not received prior treatment Download : Download high-res image (166KB) Download : Download full-size image Disclosures: Wierda: Abbott Laboratories: Research Funding. O'Brien: Avila: Research Funding; Bayer: Consultancy; Bristol-Myers Squibb: Research Funding; Gilead Sciences: Consultancy, Research Funding; Celgene: Consultancy; Cephalon: Consultancy; CII Global Research Foundation: Membership on an entity's Board of Directors or advisory committees; Genentech BioOncology: Research Funding; Genzyme: Consultancy; GlaxoSmithKline: Consultancy; MorphoSys: Consultancy; Novartis: Consultancy; Pharmacyclics: Membership on an entity's Board of Directors or advisory committees, Research Funding; Pfizer: Consultancy; Seattle Genetics, Inc.: Consultancy; Sigma Tau Pharmaceuticals: Consultancy; Talon: Research Funding; The Medal Group: Speakers Bureau.
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cd79b expression in chronic lymphocytic leukemia
Archives of Pathology & Laboratory Medicine, 2009Co-Authors: Ellen J Schlette, Jeffrey L Medeiros, Michael J. KeatingAbstract:Abstract Context.—CD79b is a relatively newly characterized B-cell marker that is expressed in a minority of chronic lymphocytic leukemia (CLL) cases. Objective.—To systematically correlate CD79b expression with specific morphologic and immunophenotypic findings and Trisomy 12. Design.—We assessed CD79b expression in 100 consecutively accrued CLL cases that were also analyzed by conventional cytogenetics. Based on the association between Trisomy 12 and CD79b expression, we then assessed 43 additional CLL cases with Trisomy 12. CD79b expression was correlated with morphology and expression of other immunophenotypic markers. Results.—Eighteen (18%) of 100 consecutively accrued cases were CD79b positive. No significant association was found between CD79b expression and atypical morphology. CD79b expression correlated with CD22 and FMC7 positivity. Eight (8%) cases had Trisomy 12; 4 (50%) of these were CD79b positive, suggesting an association with Trisomy 12. Examination of a second group of 51 CLL cases wit...
Michael J Gaffey - One of the best experts on this subject based on the ideXlab platform.
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ovarian granulosa cell tumors with bizarre nuclei an immunohistochemical analysis with fluorescence in situ hybridization documenting Trisomy 12 in the bizarre component corrected
Modern Pathology, 1996Co-Authors: Michael J Gaffey, Vandana Shashi, Christopher Von Kapherr, H F Frierson, Julia C Iezzoni, Stacey E Mills, P B Clement, D J Gersell, Robert H YoungAbstract:Granulosa cell tumors with bizarre nuclei (GCT-BN) are rare lesions with a prognosis apparently similar to that of conventional granulosa cell tumors (GCT-NOS). The immunohistochemical features of GCT-BN have not been described, and the exact nature of the bizarre nuclei (BN) is unclear. Thirteen GCT-BN were studied with antibodies to cytokeratin, vimentin, epithelial membrane antigen, muscle-specific actin, alpha smooth muscle actin, desmin, and S-100 protein. Six cases were also examined by fluorescence in situ hybridization for Trisomy 12, a nonrandom chromosomal aberration found in a proportion of ovarian sex-cord stromal tumors. Histologically, 12 tumors (86%) contained BN areas interspersed with large areas of GCT-NOS. The remaining tumor contained only microscopic foci of GCT-NOS. Immunohistochemically, the tumors stained for vimentin (13 tumors), S-100 protein (11 tumors), muscle-specific actin (10 tumors), cytokeratin (eight tumors), alpha smooth muscle actin (eight tumors), and desmin (one tumor), but none stained for epithelial membrane antigen. Immunostaining results for the BN and GCT-NOS areas were concordant in eight (73%) of the 11 tumors in which both areas could be independently assessed. The remaining three tumors (27%) showed discordant results for only one of the eight markers used. In five patients, Trisomy 12 was detected by fluorescence in situ hybridization in areas of BN but not in areas of GCT-NOS present in the same tumor. Trisomy 12 was also present in another BN tumor in which the foci of GCT-NOS were too small to be evaluated. We conclude that within GCT-BN, areas with BN are immunohistochemically similar to areas of GCT-NOS present in the same tumor. The finding of Trisomy 12 in areas with BN but not GCT-NOS in the same tumor, however, suggests that cells with BN represent a genetically distinct clone of tumor cells arising within GCT-NOS.
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interphase fluorescence in situ hybridization for Trisomy 12 on archival ovarian sex cord stromal tumors
Gynecologic Oncology, 1994Co-Authors: Vandana Shashi, Wendy L Golden, Christopher Von Kapherr, William A Andersen, Michael J GaffeyAbstract:Trisomy 12 is a nonrandom chromosomal abnormality found in a large proportion of ovarian sex cord-stromal tumors (OSCTs), including thecoma-fibromas (TFs) and granulosa cell tumors (GCTs). The prognostic significance of Trisomy 12 in these tumors, however, is unknown. A series of 16 OSCTs, obtained from patients with long-term follow-up, was analyzed for the presence of Trisomy 12 by interphase fluorescence in situ hybridization on paraffin-embedded sections. Sections of the contralateral nonneoplastic ovary were available in five cases and utilized as controls. Evidence of Trisomy 12 was detected in 9 of 10 TFs, and contrary to previous reports, in only one of six GCTs. One TF with Trisomy 12 was a malignant variant that resulted in the death of the patient in 5 months, but the remaining TFs with Trisomy 12 were cytologically and clinically benign in those with follow-up available. The single GCT with Trisomy 12 was a nonaggressive, stage 1 lesion without evidence of recurrence after 264 months, whereas those GCTs without Trisomy 12 included one stage 2 tumor and a cytologically atypical GCT with tumor necrosis and an elevated number of mitotic figures. The evidence suggests that the great majority of OSCTs with Trisomy 12 is clinically benign, but not all benign OSCTs have Trisomy 12. We conclude that the presence of Trisomy 12 is of limited prognostic usefulness in OSCTs.
E Matutes - One of the best experts on this subject based on the ideXlab platform.
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fish analysis for bcl 1 rearrangements and Trisomy 12 helps the diagnosis of atypical b cell leukaemias
Leukemia, 1999Co-Authors: E Matutes, P Carrara, L J A Coignet, Vasantha Britobabapulle, N Villamor, A Wotherspoon, Daniel CatovskyAbstract:We have investigated the diagnostic value of fluorescence in situ hybridisation (FISH) to detect t(11;14) and Trisomy 12 in 53 cases with a B cell leukaemia difficult to classify on clinical and laboratory grounds. These cases were initially diagnosed by morphology and immunophenotype and in 33 of them, on tissue histology, as follows: chronic lymphocytic leukaemia (CLL), 20, 18 of them with atypical features; B cell prolymphocytic leukaemia (B-PLL), two; mantle-cell lymphoma (MCL), 15; splenic lymphoma with villous lymphocytes (SLVL), five; lymphoplasmacytic lymphoma, six; follicular lymphoma, one and, four cases remained unclassifiable. FISH demonstrated BCL-1 rearrangement in the circulating cells from 15 cases classified as: MCL (10), atypical CLL (three) and B-PLL (two). A definitive diagnosis of MCL was made on review of the spleen histology in one out of the three atypical CLL with BCL-1 rearrangement. Trisomy 12 was detected in eight cases which included four atypical CLL, one typical CLL, two MCL and one unspecified B cell lymphoma by histology and morphology. One of the MCL had both Trisomy 12 and BCL-1 rearrangement and the other was CD5+, CD23+ and had a CLL score of 3, suggesting the latter diagnosis. Our findings demonstrate that FISH analysis is useful to clarify the nature of the disease in patients presenting with a B cell leukaemia in which the diagnosis is difficult by conventional methods. FISH established with certainty the diagnosis of MCL by showing BCL-1 rearrangement in over two-thirds of cases in which this was suspected, including blastoid forms, and confirmed the diagnosis of most cases of atypical CLL.
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atypical lymphocyte morphology an adverse prognostic factor for disease progression in stage a cll independent of Trisomy 12
British Journal of Haematology, 1997Co-Authors: David Oscier, Daniel Catovsky, E Matutes, R Gillingham, Adrian Copplestone, R M Pickering, R Chapman, Terry J HamblinAbstract:Summary. We studied 270 patients with Binet stage Achronic lymphocytic leukaemia looking for adverse prog-nostic factors. In a multivariate analysis the followingfeatures were found to be risk factors for disease progression:atypical lymphocyte morphology (defined as either >10%prolymphocytes or >15% lymphocytes with cleaved nucleior lymphoplasmacytoid cells); more than two karyotypicabnormalities; lymphocyte count >30 ×10 9 /l; lymphocytedoubling time <1 year; enlargement of one or more lymphnode groups.In a univariate analysis the presence of Trisomy 12 alsocorrelated with progressive disease, but this was largely aconsequence of the association between Trisomy 12 andatypical lymphocyte morphology. Atypical lymphocytemorphology is an important prognostic factor in stageA CLL, and one which incurs no additional investigationalcost. Keywords: atypical lymphocyte morphology, stage A CLL.The Binet and Rai staging systems separate patients withchronic lymphocytic leukaemia (CLL) into low, intermediateand high risk groups, but cannot predict which low-riskpatients will develop progressive disease (Cheson, 1993;Montserrat & Rozman, 1993). Until recently the predictionof disease progression was of scientific interest but of littlepractical importance since there was no evidence thattreatment of Binet stage A patients with alkylating agentsbefore progression occurred was of any benefit (FrenchCo-operative Group on CLL, 1990). However, there is nowconsiderable interest in the use of new and more intensivetreatments such as nucleoside analogues (Keating
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correlation of Trisomy 12 with proliferating cells by combined immunocytochemistry and fluorescence in situ hybridization in chronic lymphocytic leukemia
Leukemia, 1996Co-Authors: J A Garciamarco, Cathy M Price, J Ellis, E Matutes, M Morey, Daniela Lens, Susan M Colman, D CatovskyAbstract:Conventional G-banding and fluorescence in situ hybridization (FISH) were performed on peripheral blood samples of 340 consecutive untreated cases of chronic lymphocytic leukemia (CLL) for the detection of Trisomy 12 and other chromosome abnormalities. These findings were correlated with the proliferative activity of CLL lymphocytes assessed by the monoclonal antibody Ki-67. Cytogenetic analysis displayed a normal karyotype in 131 (38.5%) cases, Trisomy 12 in 68 (20%), 31 by G-banding and an additional 37 cases by FISH, other clonal abnormalities in 47 (14%), and no metaphases in 94 (27.5%). The percentage of Ki-67-positive cells was significantly higher in cases with Trisomy 12 (4.1 +/- 4.48) than in cases with a normal karyotype (1.5 +/- 2.0), those with other clonal abnormalities (1.35 +/- 1.37) and cases with no metaphases (1.14 +/- 1.6) (P< 0.0001). Cases with Trisomy 12 were associated with more advanced clinical stage, atypical morphology and a higher percentage of Ki-67+ve cells than cases lacking Trisomy 12 (P< 0.0001). Although there was no direct correlation between the percentage of trisomic and proliferating cells, the combination of immunocytochemistry and FISH showed that most Ki-67-positive cells were trisomic for chromosome 12. Our results suggest that the association of Trisomy 12 with a higher proliferative activity supports the view that this abnormality is a secondary event associated with disease progression in CLL.
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Trisomy 12 defines a group of cll with atypical morphology correlation between cytogenetic clinical and laboratory features in 544 patients
British Journal of Haematology, 1996Co-Authors: E Matutes, David Oscier, J Ellis, J Garciamarco, A Copplestone, R Gillingham, Terry J Hamblin, D Lens, G J Swansbury, D CatovskyAbstract:We have analysed the clinical and laboratory features in 544 patients with chronic lymphocytic leukaemia (CLL) with available cytogenetics and fluorescence in-situ hybridization (FISH) analysis for Trisomy 12 in half of them, to examine the correlation between chromosome abnormalities and clinical or laboratory parameters. Five chromosome groups were defined: (1) Trisomy 12 (18%), detected as the sole abnormality or associated with other changes; (2) del(13)(q12-14) (7%); (3) other abnormal karyotypes (20%); (4) normal karyotype (41%); and (5) no divisions (14%). There were no differences in the age distribution between the five groups. Clinical stages (Binet) were: A (74%), B (12%) and C (14%). Stage A was common in cases with del(13q)(82%), normal (84%) and other abnormal karyotypes (74%), whereas it was less common in Trisomy 12 cases (64%) and those with no divisions (48%). Typical CLL morphology was found in 83% of cases; 10% had more than 10% prolymphocytes (CLL/PL) and 7% had other atypical features. CLL with Trisomy 12 was the only group with a high frequency of either CLL/PL (31%) or atypical morphology (24%). Atypical morphology and CLL/PL were even more frequent when Trisomy 12 was associated with other chromosomal abnormalities (70% v 46%). The incidence of cases with CLL/PL and other atypical morphology was significantly lower in the other chromosome groups (P < 0.001). There were no differences in immunophenotype among the various groups except for a higher frequency of stronger Smlg and FMC7 expression in cases with Trisomy 12, particularly those with CLL/PL and other atypical morphology. Our findings confirm that Trisomy 12 defines a subgroup of CLL with more frequent atypical morphology, including CLL/PL, stronger SmIg and FMC7 expression, more advanced stages (B and C in 18%) and possibly worse prognosis.
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Trisomy 12 in b cell chronic lymphocytic leukaemia assessment of lineage restriction by simultaneous analysis of immunophenotype and genotype in interphase cells by fluorescence in situ hybridization
British Journal of Haematology, 1994Co-Authors: J Garcigamarco, David Oscier, J Ellis, E Matutes, D Catovsky, R Morilla, J Fantes, Cathy M PriceAbstract:Summary. We have studied the lineage restriction of Trisomy 12 in six patients with B-cell chronic lymphocytic leukaemia (CLL) by simultaneous analysis of immunophenotype and fluorescence in situ hybridization (FISH) signals in single interphase cells. Fresh uncultured cells from each patient were immunophenotyped by the alkaline phosphatase anti-alkaline phosphatase method (APAAP) using monoclonal or polyclonal antibodies and hybridized with a chromosome 12 specific alpha-satellite DNA probe. In all cases Trisomy 12 was restricted to the clonal B-cells, kappa positive or lambda positive, whereas T-cells (CD3 positive) and non clonal B-cells had only two chromosome 12 signals. Within the clonal B-cell population a large proportion of cells were disomic for chromosome 12, whilst trisomic cells ranged from 21% to 37%. The absence of Trisomy 12 in T-cells and the mosaicism demonstrated in the clonal B-cells suggests that this abnormality is a secondary event during the leukaemic transformation of CLL and develops in an already established neoplastic B-cell population.
Vandana Shashi - One of the best experts on this subject based on the ideXlab platform.
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ovarian granulosa cell tumors with bizarre nuclei an immunohistochemical analysis with fluorescence in situ hybridization documenting Trisomy 12 in the bizarre component corrected
Modern Pathology, 1996Co-Authors: Michael J Gaffey, Vandana Shashi, Christopher Von Kapherr, H F Frierson, Julia C Iezzoni, Stacey E Mills, P B Clement, D J Gersell, Robert H YoungAbstract:Granulosa cell tumors with bizarre nuclei (GCT-BN) are rare lesions with a prognosis apparently similar to that of conventional granulosa cell tumors (GCT-NOS). The immunohistochemical features of GCT-BN have not been described, and the exact nature of the bizarre nuclei (BN) is unclear. Thirteen GCT-BN were studied with antibodies to cytokeratin, vimentin, epithelial membrane antigen, muscle-specific actin, alpha smooth muscle actin, desmin, and S-100 protein. Six cases were also examined by fluorescence in situ hybridization for Trisomy 12, a nonrandom chromosomal aberration found in a proportion of ovarian sex-cord stromal tumors. Histologically, 12 tumors (86%) contained BN areas interspersed with large areas of GCT-NOS. The remaining tumor contained only microscopic foci of GCT-NOS. Immunohistochemically, the tumors stained for vimentin (13 tumors), S-100 protein (11 tumors), muscle-specific actin (10 tumors), cytokeratin (eight tumors), alpha smooth muscle actin (eight tumors), and desmin (one tumor), but none stained for epithelial membrane antigen. Immunostaining results for the BN and GCT-NOS areas were concordant in eight (73%) of the 11 tumors in which both areas could be independently assessed. The remaining three tumors (27%) showed discordant results for only one of the eight markers used. In five patients, Trisomy 12 was detected by fluorescence in situ hybridization in areas of BN but not in areas of GCT-NOS present in the same tumor. Trisomy 12 was also present in another BN tumor in which the foci of GCT-NOS were too small to be evaluated. We conclude that within GCT-BN, areas with BN are immunohistochemically similar to areas of GCT-NOS present in the same tumor. The finding of Trisomy 12 in areas with BN but not GCT-NOS in the same tumor, however, suggests that cells with BN represent a genetically distinct clone of tumor cells arising within GCT-NOS.
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interphase fluorescence in situ hybridization for Trisomy 12 on archival ovarian sex cord stromal tumors
Gynecologic Oncology, 1994Co-Authors: Vandana Shashi, Wendy L Golden, Christopher Von Kapherr, William A Andersen, Michael J GaffeyAbstract:Trisomy 12 is a nonrandom chromosomal abnormality found in a large proportion of ovarian sex cord-stromal tumors (OSCTs), including thecoma-fibromas (TFs) and granulosa cell tumors (GCTs). The prognostic significance of Trisomy 12 in these tumors, however, is unknown. A series of 16 OSCTs, obtained from patients with long-term follow-up, was analyzed for the presence of Trisomy 12 by interphase fluorescence in situ hybridization on paraffin-embedded sections. Sections of the contralateral nonneoplastic ovary were available in five cases and utilized as controls. Evidence of Trisomy 12 was detected in 9 of 10 TFs, and contrary to previous reports, in only one of six GCTs. One TF with Trisomy 12 was a malignant variant that resulted in the death of the patient in 5 months, but the remaining TFs with Trisomy 12 were cytologically and clinically benign in those with follow-up available. The single GCT with Trisomy 12 was a nonaggressive, stage 1 lesion without evidence of recurrence after 264 months, whereas those GCTs without Trisomy 12 included one stage 2 tumor and a cytologically atypical GCT with tumor necrosis and an elevated number of mitotic figures. The evidence suggests that the great majority of OSCTs with Trisomy 12 is clinically benign, but not all benign OSCTs have Trisomy 12. We conclude that the presence of Trisomy 12 is of limited prognostic usefulness in OSCTs.
David Oscier - One of the best experts on this subject based on the ideXlab platform.
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atypical lymphocyte morphology an adverse prognostic factor for disease progression in stage a cll independent of Trisomy 12
British Journal of Haematology, 1997Co-Authors: David Oscier, Daniel Catovsky, E Matutes, R Gillingham, Adrian Copplestone, R M Pickering, R Chapman, Terry J HamblinAbstract:Summary. We studied 270 patients with Binet stage Achronic lymphocytic leukaemia looking for adverse prog-nostic factors. In a multivariate analysis the followingfeatures were found to be risk factors for disease progression:atypical lymphocyte morphology (defined as either >10%prolymphocytes or >15% lymphocytes with cleaved nucleior lymphoplasmacytoid cells); more than two karyotypicabnormalities; lymphocyte count >30 ×10 9 /l; lymphocytedoubling time <1 year; enlargement of one or more lymphnode groups.In a univariate analysis the presence of Trisomy 12 alsocorrelated with progressive disease, but this was largely aconsequence of the association between Trisomy 12 andatypical lymphocyte morphology. Atypical lymphocytemorphology is an important prognostic factor in stageA CLL, and one which incurs no additional investigationalcost. Keywords: atypical lymphocyte morphology, stage A CLL.The Binet and Rai staging systems separate patients withchronic lymphocytic leukaemia (CLL) into low, intermediateand high risk groups, but cannot predict which low-riskpatients will develop progressive disease (Cheson, 1993;Montserrat & Rozman, 1993). Until recently the predictionof disease progression was of scientific interest but of littlepractical importance since there was no evidence thattreatment of Binet stage A patients with alkylating agentsbefore progression occurred was of any benefit (FrenchCo-operative Group on CLL, 1990). However, there is nowconsiderable interest in the use of new and more intensivetreatments such as nucleoside analogues (Keating
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differential rates of somatic hypermutation in vh genes among subsets of chronic lymphocytic leukemia defined by chromosomal abnormalities
Blood, 1997Co-Authors: David Oscier, Freda K Stevenson, Andrew R Thompsett, Delin ZhuAbstract:Chronic lymphocytic leukemia (CLL) is a B-cell tumor involving small lymphocytes that generally express the CD5 antigen and low levels of surface Ig. Within this definition, there is heterogeneity among cases in cell morphology, karyotypic abnormalities, and clinical course. Trisomy 12, the most frequent karyotypic abnormality, is commonly found in a subset of CLL with atypical morphology. It has also been associated with advanced disease, and possibly with a less favorable prognosis. A further subset of cases with abnormalities involving chromosome 13q14 have typical lymphocyte morphology. Occasionally, the two abnormalities are found together. To assess the clonal history of the cell of origin in disease subsets defined by these two chromosomal abnormalities, we investigated the usage of VH genes and the pattern of somatic mutation in 10 cases of Trisomy 12 with atypical morphology and eight cases of 13q14 abnormality with typical morphology. In addition, four cases with both chromosomal abnormalities were analyzed. Results confirm a common usage of the VH1 family in all subsets. However, the patterns of somatic mutation were distinct, with cases of Trisomy 12 showing a minimal level of mutation (mean ± SD, 0.34% ± 0.86%) and cases of 13q14 abnormality showing significant levels (6.5% ± 1.67%). The four cases with both abnormalities showed a mixed pattern. All mutated cases had intraclonal homogeneity, and three of 10 had a pattern indicative of antigen selection. These results suggest that the clonal history of the two subsets of CLL may differ.
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Trisomy 12 and structural abnormalities of 13q14 occurring in the same clone in chronic lymphocytic leukaemia
British Journal of Haematology, 1996Co-Authors: Sarah Mould, Anne Gardiner, Martin Corcoran, David OscierAbstract:Trisomy 12 and deletions or translocations of 13q14 are the commonest cytogenetic abnormalities in chronic lymphocytic leukaemia but rarely co-exist in the same patient. We describe eight patients from a series of > 400 patients with CLL in whom Trisomy 12 and t or del 13 occur in the same clone. Using FISH we have identified clones with Trisomy 12 alone, t or del 13q14 alone and both abnormalities, in each of the patients studied. This implies that neither Trisomy 12 nor t or del 13q14 is the initiating event in leukaemogenesis, but does not exclude the possibility of a submicroscopic abnormality of 13q14 occurring as an early event.
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Trisomy 12 defines a group of cll with atypical morphology correlation between cytogenetic clinical and laboratory features in 544 patients
British Journal of Haematology, 1996Co-Authors: E Matutes, David Oscier, J Ellis, J Garciamarco, A Copplestone, R Gillingham, Terry J Hamblin, D Lens, G J Swansbury, D CatovskyAbstract:We have analysed the clinical and laboratory features in 544 patients with chronic lymphocytic leukaemia (CLL) with available cytogenetics and fluorescence in-situ hybridization (FISH) analysis for Trisomy 12 in half of them, to examine the correlation between chromosome abnormalities and clinical or laboratory parameters. Five chromosome groups were defined: (1) Trisomy 12 (18%), detected as the sole abnormality or associated with other changes; (2) del(13)(q12-14) (7%); (3) other abnormal karyotypes (20%); (4) normal karyotype (41%); and (5) no divisions (14%). There were no differences in the age distribution between the five groups. Clinical stages (Binet) were: A (74%), B (12%) and C (14%). Stage A was common in cases with del(13q)(82%), normal (84%) and other abnormal karyotypes (74%), whereas it was less common in Trisomy 12 cases (64%) and those with no divisions (48%). Typical CLL morphology was found in 83% of cases; 10% had more than 10% prolymphocytes (CLL/PL) and 7% had other atypical features. CLL with Trisomy 12 was the only group with a high frequency of either CLL/PL (31%) or atypical morphology (24%). Atypical morphology and CLL/PL were even more frequent when Trisomy 12 was associated with other chromosomal abnormalities (70% v 46%). The incidence of cases with CLL/PL and other atypical morphology was significantly lower in the other chromosome groups (P < 0.001). There were no differences in immunophenotype among the various groups except for a higher frequency of stronger Smlg and FMC7 expression in cases with Trisomy 12, particularly those with CLL/PL and other atypical morphology. Our findings confirm that Trisomy 12 defines a subgroup of CLL with more frequent atypical morphology, including CLL/PL, stronger SmIg and FMC7 expression, more advanced stages (B and C in 18%) and possibly worse prognosis.
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Trisomy 12 in b cell chronic lymphocytic leukaemia assessment of lineage restriction by simultaneous analysis of immunophenotype and genotype in interphase cells by fluorescence in situ hybridization
British Journal of Haematology, 1994Co-Authors: J Garcigamarco, David Oscier, J Ellis, E Matutes, D Catovsky, R Morilla, J Fantes, Cathy M PriceAbstract:Summary. We have studied the lineage restriction of Trisomy 12 in six patients with B-cell chronic lymphocytic leukaemia (CLL) by simultaneous analysis of immunophenotype and fluorescence in situ hybridization (FISH) signals in single interphase cells. Fresh uncultured cells from each patient were immunophenotyped by the alkaline phosphatase anti-alkaline phosphatase method (APAAP) using monoclonal or polyclonal antibodies and hybridized with a chromosome 12 specific alpha-satellite DNA probe. In all cases Trisomy 12 was restricted to the clonal B-cells, kappa positive or lambda positive, whereas T-cells (CD3 positive) and non clonal B-cells had only two chromosome 12 signals. Within the clonal B-cell population a large proportion of cells were disomic for chromosome 12, whilst trisomic cells ranged from 21% to 37%. The absence of Trisomy 12 in T-cells and the mosaicism demonstrated in the clonal B-cells suggests that this abnormality is a secondary event during the leukaemic transformation of CLL and develops in an already established neoplastic B-cell population.
