scholarly journals Evaluation of MLPA as a comprehensive molecular cytogenetic tool to detect cytogenetic markers of chronic lymphocytic leukemia in Egyptian patients

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Ola M. Eid ◽  
Rania M. A. Abdel Kader ◽  
Lamiaa A. Fathalla ◽  
Amany H. Abdelrahman ◽  
Ahmed Rabea ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Chromosomal Aberrations ◽  
Cost Effective ◽  
Lymphocytic Leukemia ◽  
Fish Analysis ◽  
Molecular Cytogenetic ◽  
Diagnosis And Prognosis ◽  
Trisomy 12 ◽  
Screening Platform ◽  
Adult Leukemia

Abstract Background Chronic lymphocytic leukemia (CLL) is the most common form of adult leukemia. This disease is genetically heterogeneous, and approximately 85% of patients with CLL harbor chromosomal aberrations that are considered effective prognostic biomarkers. The most frequent aberrations include deletions in 13q14, followed by trisomy 12, and deletions in 11q22.3 and 17p13 (TP53). Currently, fluorescence in situ hybridization (FISH) is the most widely used molecular cytogenetic technique to detect these aberrations. However, FISH is laborious, time-consuming, expensive, and has a low throughput. In contrast, multiplex ligation-dependent probe amplification (MLPA) is a reliable, cost-effective, and relatively rapid technique that can be used as a first-line screening tool and complement with FISH analysis. This study aimed to evaluate the contributions of MLPA as a routine standalone screening platform for recurrent chromosomal aberrations in CLL in comparison to other procedures. Thirty patients with CLL were screened for the most common genomic aberrations using MLPA with SALSA MLPA probemix P038-B1 CLL and FISH. Results In 24 of the 30 cases (80%), the MLPA and FISH results were concordant. Discordant results were attributed to a low percentage of mosaicism. Moreover, the MLPA probemix contains probes that target other genomic areas known to be linked to CLL in addition to those targeting common recurrent CLL aberrations. Conclusions The usage of MLPA as the first screening platform followed by FISH technique for only the negative cases is the most appropriate approach for CLL diagnosis and prognosis.

scholarly journals A high-resolution allelotype of B-cell chronic lymphocytic leukemia (B-CLL)

Blood ◽  
2002 ◽  
Vol 100 (5) ◽  
pp. 1787-1794 ◽  
Author(s):  
Urban Novak ◽  
Elisabeth Oppliger Leibundgut ◽  
Jörg Hager ◽  
Dominique Mühlematter ◽  
Martine Jotterand ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Chromosomal Aberrations ◽  
Cytogenetic Analysis ◽  
Prognostic Significance ◽  
Genetic Alterations ◽  
Lymphocytic Leukemia ◽  
Accurate Information ◽  
Trisomy 12 ◽  
Cell Chronic Lymphocytic Leukemia

The most frequent chromosomal aberrations in B-cell chronic lymphocytic leukemia (B-CLL) are deletions on 13q, 11q, and 17p, and trisomy 12, all of which are of prognostic significance. Conventional cytogenetic analysis and fluorescence in situ hybridization (FISH) are used for their detection, but cytogenetic analysis is hampered by the low mitotic index of B-CLL cells, and FISH depends on accurate information about candidate regions. We used a set of 400 highly informative microsatellite markers covering all chromosomal arms (allelotyping) and automated polymerase chain reaction (PCR) protocols to screen 46 patients with typical B-CLL for chromosomal aberrations. For validation, we compared data with our conventional karyotype results and fine mapping with conventional single-site PCR. All clonal cytogenetic abnormalities potentially detectable by our microsatellite PCR (eg, del13q14 and trisomy 12) were picked up. Allelotyping revealed additional complex aberrations in patients with both normal and abnormal B-CLL karyotypes. Aberrations detectable in the samples with our microsatellite panel were found on almost all chromosomal arms. We detected new aberrant loci in typical B-CLL, such as allelic losses on 1q, 9q, and 22q in up to 25% of our patients, and allelic imbalances mirroring chromosomal duplications, amplifications, or aneuploidies on 2q, 10p, and 22q in up to 27% of our patients. We conclude that allelotyping with our battery of informative microsatellites is suitable for molecular screening of B-CLL. The technique is well suited for analyses in clinical trials, it provides a comprehensive view of genetic alterations, and it may identify new loci with candidate genes relevant in the molecular biology of B-CLL.

Incidence of Genomic Aberrations and Associated Efficacy from a Phase III Study Comparing Alemtuzumab (CAMPATH®, MABCAMPATH®) vs Chlorambucil as First Line Therapy for B-Cell Chronic Lymphocytic Leukemia (BCLL).

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2092-2092 ◽  
Author(s):  
Tadeusz Robak ◽  
Anna Dmoszynska ◽  
Raouf Fetni ◽  
Ying Wang ◽  
Malika Belkacz ◽  
...  
Keyword(s):  
Chromosomal Aberrations ◽  
Small Sample Size ◽  
Variable Region ◽  
Small Sample ◽  
Lymphocytic Leukemia ◽  
Phase Iii ◽  
Chain Gene ◽  
Fish Analysis ◽  
Myc Oncogene ◽  
Trisomy 12

Abstract CAM307 is a randomized Phase III trial comparing the efficacy and safety of alemtuzumab (CAM) with chlorambucil (CHLO). The trial enrolled 297 previously untreated patients (pts) requiring therapy according to NCI-WG criteria. Pts were randomized 1:1 to CAM (n=149) vs CHLO (n=148) using standard dosing regimens. Fluorescence in situ hybridization (FISH) on interphase nuclei of lymphocytes isolated from blood was analyzed for cytogenetic abnormalities prior to the start of therapy. FISH analysis was performed using 13 DNA probes to detect chromosomal aberrations in 17p13.1 (P53), 13q14 (RB1, D13S319 and D13S25), 11q22.3-11q23 (ATM and MLL), 6q27 (subtelomere), 6q21 (chromosome 6q21/alphasatellite 6 cocktail probe), trisomy 8q24 (c-myc), trisomy 12 (CEP12) and translocations involving the locus of immunoglobulin heavy chain gene (IGH, 14q32.33). Samples were analyzed in 282 pts (95%); chromosomal aberrations were detected in 231 pts (82%) while 51 pts (18%) exhibited a normal interphase FISH pattern. The most frequent abnormalities were deletions (del) at loci 13q (49%), sole del 13q (24%), 11q (19%), 17p (7 %), 6q (4 %), and trisomies 12 (14%) and 8q (5%). Translocations IGH, 14q32.33 were detected in 10 pts (4%). An exploratory analysis was performed to correlate time to event variables (assessed by an independent response review panel) with cytogenetics. Overall 165 pts (59%) revealed combination abnormalities. The most frequently observed chromosomal associations were: del 13q + del 14q (N=20, 12%), del 11q + del 13q (N=17, 10%), del 11q + del 13q + del 14q (N=11, 7%), del 11q + del 14q (N=7, 4%), trisomy 12 + del 13q (N=5, 3%), del 13q + del 17p (N=4, 2%), del 11q + trisomy 12 (N=3, 2%) and del 17p + del 6q (N=3, 2%). Coexistence of del 17p and del 11q was not observed. Although del 13q was observed with all chromosome abnormalities, nearly half of the cases del 13q14.3 (D13S25 and D13S319) coincided with an ATM deletion (11q22.3). FISH analysis has allowed the detection of uncommon abnormalities: tetraploidy (n=1), hyperdiploidy (n=1), trisomy 18 (n=1) and c-myc oncogene amplification (>15 copies per nuclei) (n=2). The latter is a well known abnormality in solid tumors but rarely seen in leukemia. In addition, del of the IGH variable region was detected in 70 pts. The biological and clinical significance of this abnormality is to be investigated. Conclusions: Overall, 82% of treatment naïve BCLL pts revealed cytogenetic aberrations and 59% were combination abnormalities. CAM307 demonstrates a significant improvement in PFS in pts treated with CAM vs CHLO who present with del 13q as the sole abnormality; no difference in pts with del 11q. However, a trend towards improved PFS was observed in pts with trisomy 12 and del 17p, which did not reach significance due to small sample size. Further investigation of CAM therapy in high risk cytogenetic subgroups is warranted.

Influence of Chromosomal Aberrations on Response to First Line Therapy in B-Cell Chronic Lymphocytic Leukemia: Interim Analysis of PALG-CLL3 Randomized Study Comparing Cladribine Plus Cyclophosphamide vs Fludarabine Plus Cyclophosphamide.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2046-2046
Author(s):  
Tadeusz Robak ◽  
Jerzy Z. Blonski ◽  
Ewa Wawrzyniak ◽  
Aleksandra Palacz ◽  
Joanna Gora-Tybor ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Chromosomal Aberrations ◽  
Interim Analysis ◽  
Chromosomal Abnormalities ◽  
Complete Response ◽  
Lymphocytic Leukemia ◽  
Lower Probability ◽  
Trisomy 12 ◽  
Cell Chronic Lymphocytic Leukemia

Abstract Impact of cytogenetic abnormalities on treatment with different purine nucleoside analogs in patients (pts) with B-cell chronic lymphocytic leukemia (B-CLL) is largely unknown. One of objectives of PALG-CLL3 trial, comparing cladribine plus cyclophosphamide (CC) with fludarabine plus cyclophosphamide (FC) in previously untreated progressive B-CLL, was to verify the response to treatment in subsets of pts characterized by common cytogenetic abberrations. Chromosomal abnormalities were assessed using fluorescence in situ hybridization (FISH) on interphase nuclei of lymphocytes on whole blood smears prior to the start of the study treatment. Pts were screened for trisomy 12, deletions (del) 11q, del 13q and del 17p using DNA probes: CEP12, LSI: ATM, D13S319 and p53 (Vysis), respectively. For the purpose of the present interim analysis complete cytogenetic results were available in 133 pts out of 423 pts included to the study. In this group the chromosomal aberrations were detected in 102 pts (77%) including single abnormalities observed in 69 pts (52%) and two or more aberrations in 33 pts (25%). Thirty-one pts (23%) exhibited a normal interphase FISH pattern. The most frequent single abnormality was del 13q found in 38 pts (29%), while del 17p, trisomy 12 and del 11q were identified in 14 pts (11%), 11 pts (8%), and 6 pts, (5%), respectively. The most frequently observed associations of chromosomal aberrations were: del 13q with del 11q (11 pts, 8%) and del 13q with del 17p (10 pts, 8%). Four pts (3%) revealed three chromosomal abnormalities including association of trisomy 12/del 11q/del 13q in two pts, trisomy 12/del 11q/del 17p in one pt and del 11q/del 13q/del 17p in one pt. Overall, treatment was completed and response assessed in 113 out of 133 pts with known FISH pattern. In this group of pts del 17p was the only chromosomal abnormality that correlated significantly with treatment outcome. Pts with del 17p (21, 19%) had lower probability to achieve a complete response (CR) (0.044). Interestingly, in independent analyses of both treatment arms, the negative impact of 17p was seen in pts treated with FC (p=0.002), but not in pts treated with CC (p=0.6). Moreover, comparing response rates between treatment arms we found that CC was superior to FC in terms of complete response in pts with del 17p (57% CR in CC v 14% CR in FC arm, p=0.04). In conclusion, chromosomal abnormalities can be detected in majority B-CLL pts requiring treatment. Our preliminary results suggest that CC combination may have some advantage in terms of CR achievement in B-CLL pts harboring del 17p.

Normal Fish Cytogenetics and 13q Deletions Unveil Marked Biological and Clinical Heterogeneity In Chronic Lymphocytic Leukemia

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2692-2692
Author(s):  
Giovanni Del Poeta ◽  
Maria Ilaria Del Principe ◽  
Francesca Maria Rossi ◽  
Angela Coletta ◽  
Francesco Buccisano ◽  
...  
Keyword(s):  
Prognostic Factor ◽  
Chronic Lymphocytic Leukemia ◽  
Independent Prognostic Factor ◽  
Lymphocytic Leukemia ◽  
Fish Analysis ◽  
Fish Cytogenetics ◽  
Cd69 Expression ◽  
Trisomy 12 ◽  
Normal Fish ◽  
Normal Cytogenetics

Abstract Abstract 2692 Cytogenetic aberrations are considered major prognostic indicators for predicting the survival of chronic lymphocytic leukemia (CLL) patients (pts) [Dohner et al, 2000]. Given the difficulties in obtaining abnormal metaphases in CLL, fluorescent in situ hybridization (FISH) with specific probes is generally used to detect the most frequent abnormalities. Interphase FISH (I-FISH) is a rapid and sensitive technique for analysis of chromosome aberrations in CLL, but the limited number of DNA probes available to screen B-CLL results in a high number of cases presenting normal cytogenetics. Moreover, deletion 13q14 on FISH analysis which is the most common cytogenetic abnormality in CLL is a favourable prognostic biomarker when detected as a sole abnormality, but a higher percentage of 13q- nuclei was found to be associated with significantly shorter time to treatment (P<0.001), [van Dike et al, 2010]. On this line, the primary endpoints of our research were: 1) to determine progression free survival (PFS) and overall survival (OS) within the normal FISH cytogenetics subset on the basis both of ZAP-70, CD38, CD69 expression by flow cytometry and IgVH status; 2) to correlate percentages of 13q- nuclei (<50% or ≥50%) with PFS and OS and finally 3) to confirm I-FISH as an independent prognostic factor. We investigated 400 pts, median age 65 years (range 33–89), 235 males and 165 females. With regard to modified Rai stages, 115 pts had a low stage, 266 an intermediate stage and 19 a high stage. One hundred forty-two pts (35.5%) exhibit a normal karyotype, 133 pts (33.2%) showed an isolated 13q-, 69 pts (17.3%) presented trisomy 12, 30 pts (7.5%) 11q deletion, 20 (5%) 17p deletion and 6 (1.5%) other chromosome abnormalities. Clearly, pts with intermediate/poor cytogenetic abnormalities (trisomy 12, del11q-, del17p-) showed significant shorter PFS and OS (8% vs 33% at 16 years, P<0.0001 and 41% vs 65% at 16 years, P=0.0001) in comparison with normal or del13q- pts. Therefore, in order to better define the apparently homogenous subset of the patients with normal cytogenetics, we analysed ZAP-70, CD38, CD69 and IgVH status within this subgroup. Interestingly, we observed significant longer PFS and OS for ZAP-70 negative pts (60% vs 29% at 10 years, P<0.0001 and 92% vs 23% at 16 years, P=0.0006), for CD38 negative pts (54% vs 22% at 10 years, P=0.0008 and 83% vs 0% at 14 years, P=0.03), for CD69 negative pts (60% vs 23% at 10 years, P=0.0003 and 83% vs 63% at 14 years, P=0.02) and finally also for IgVH mutated cases (64% vs 35% at 10 years, P=0.0002 and 95% vs 45% at 14 years, P=0.04). Moreover, pts with isolated 13q- in <50% of nuclei (60 pts) showed a longer PFS and OS (68% vs 6% at 16 years, P=0.0003 and 92% vs 0% at 18 years, P=0.002; Figure) compared to those with ≥50% of nuclei (72 pts). No significant correlation was found between 13q- percentages and CD38 or ZAP-70 or IgVH status, while it was found with CD69 expression (P=0.01). In multivariate analysis of PFS, FISH cytogenetics was confirmed to be an independent prognostic factor (P=0.002) together with Rai stages (P=0.005), ZAP-70 (P=0.0001), IgVH status (P=0.0002) and CD69 (P=0.007). Therefore, in the clinical practice, ZAP-70, CD38, IgVH status and CD69 are irreplaceable in order to define prognosis and treatment within the large series of CLL pts with normal cytogenetics, probably presenting abnormalities undetectable by FISH. Besides, the percentage of nuclei exhibiting 13q- has to be considered an important predictor of the outcome and the clinical implications of 13q- in CLL seem to appear more complex than originally considerated. In conclusion, a comprehensive scoring prognostic system based on a combination of clinical, genetic, phenotypic parameters and IgVH status improves the separation of prognostic subgroups in CLL already early in the course of the disease. Disclosures: No relevant conflicts of interest to declare.

Flow Cytometric Identification of Biclonal Disease Among 5,523 Patients with Chronic Lymphocytic Leukemia and Its Genetic Characterization

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3608-3608
Author(s):  
Wolfgang Kern ◽  
Susanne Schnittger ◽  
Frank Dicker ◽  
Torsten Haferlach ◽  
Claudia Haferlach
Keyword(s):  
Light Chain ◽  
Chromosomal Aberrations ◽  
Molecular Genetic ◽  
Lymphocytic Leukemia ◽  
Fish Analysis ◽  
Employment Equity ◽  
Mutational Status ◽  
Trisomy 12 ◽  
Genetic Level ◽  
Equity Ownership

Abstract Abstract 3608 Chronic lymphocytic leukemia (CLL) is an indolent disease with largely heterogeneous clinical course. While the diagnosis is made based on the characteristic immunophenotype as determined by multiparameter flow cytometry (MFC) another aspect of major clinical importance is the estimation of the prognosis which includes the analysis of chromosomal aberrations, the IGHV mutational status as well as the expression of CD38 and ZAP-70. In general, the methods used to determine these parameters are applied in the assumption of analyzing one homogeneous leukemic population and are evaluated accordingly. The potential presence of subpopulations and even subclones may not always be considered adequately in this regard. We identified 76 out of 5,523 patients (1.4%) in whom MFC identified biclonal disease based on the presence of both kappa- and lambda-light chain restricted leukemic subpopulations. In 45 of these cases fluorescence in situ hybridization (FISH) analysis was performed applying a standard set of probes for the detection of del(6q), del(11q22.3) (ATM), trisomy 12, del(13q14) (D13S25, D13S319), and del(17p13) (TP53). In 17 of the 76 patients a chromosome banding analysis (CBA) was performed and the IGHV mutational status was determined in 38 of 76 patients. The patients` ages ranged from 47.7 to 88.0 years (median, 71.3 years), 50 patients were male. The median WBC count amounted to 19 ×10e9/l (range, 0.6–237 ×10e9/l). In most cases the kappa-light chain restricted subpopulation was larger than the lambda-light chain restricted one. The median values and ranges for the respective percentages of subpopulations amounted to: kappa, 24%, 1%-88%; lambda, 12%, 1%-85%. The respective peripheral blood concentrations amounted to: kappa, 3.68 ×10e9/l, 0.12–142 ×10e9/l; lambda, 2.37 ×10e9/l, 0.01–128 ×10e9/l. The median ratio kappa population/lambda population amounted to 1.9 (range, 0.05–76). FISH analysis identified del(6q) in 2/44 (4.5%) cases, del(11q) in 2/44 (4.5%), trisomy 12 in 7/44 (15.9%), del(13q) in 28/45 (62.2%), del(13q) as sole aberration detected by FISH in 23/43 (53.5%), and del(17p) in 1/45 (2.2%). In three cases more than one aberration was detected by FISH: two cases with del(11q) and del(13q) and one case with trisomy 12 and del(13q). While in two of these three cases the size-ratios of the respective subpopulations were similar in MFC and FISH analysis (1.7:1 vs. 4.3:1 and 2.0:1 vs. 1.5:1) this was not true for the third case (15.5:1 vs. 1.1:1). The further two cases could be considered in line with both methods detecting independent clones. In the latter case the chromosomal aberrations were present in 54% and 59% of the cells and the subpopulations detected by MFC amounted to 62% and 4%. Thus, both chromosomal aberrations must be considered to coexist in one population and not related to the two subpopulations detected by MFC. Overall, however, no clear-cut conclusions can be drawn from FISH results regarding the presence of independent subpopulations and therefore we next focused on the results of CBA. Within the 17 patients analyzed by CBA 12 cases showed an aberrant karyotype (70.6%). In four of these cases more than one clone was identified by differences in the chromosomal aberrations, respectively. In three cases chromosomal evolution was suggested by shared aberrations in both clones and additional aberrations in one of both clones only, respectively. Conversely, in the fourth case two completely different aberration patterns were observed. In 14 out of the 38 patients (36.8%) in whom an IGHV mutational analysis was performed two independent clones were identified by the presence of two different B-cell receptor rearrangements. The presence of biclonal disease had no impact on the clinical outcome of the patients as assessed by time to therapy and overall survival. This data indicates that subpopulations can be identified in a significant number of patients with CLL based on the immunophenotype as well as on the cytogenetic and molecular genetic level. These subpopulations at least in part must be considered as subclones with differing genetic background. These subclones may be associated with differing clinical courses. This data therefore suggests to vigorously screen patients with CLL for subpopulations by MFC and to comprehensively characterize positive cases cytogenetically by CBA and FISH analysis as well as on the molecular genetic level. Disclosures: Kern: MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Schnittger:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Dicker:MLL Munich Leukemia Laboratory: Employment. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership.

scholarly journals Coexistence of trisomy 12 and del(13)(q14.3) in two patients with chronic lymphocytic leukemia

2009 ◽  
Vol 61 (4) ◽  
pp. 607-611
Author(s):  
Marija Dencic-Fekete ◽  
D. Antic ◽  
Sanja Davidovic-Mrsic ◽  
Ivana Franic ◽  
Nada Kraguljac-Kurtovic ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Prognostic Significance ◽  
Lymphocytic Leukemia ◽  
Stable Phase ◽  
Fish Analysis ◽  
Fluo Rescence ◽  
Trisomy 12 ◽  
One Year ◽  
The Given

We describe two patients with diagnosis of chronic lymphocytic leukemia (CLL) in whom interphase fluo?rescence in situ hybridization (FISH) analysis revealed trisomy 12 and del(13)(q14.3) occurring in the same clone. These abnormalities are rarely seen together and the prognostic relevance of their coexistence is still unclear. According to some data, a probable adverse prognosis for this group of patients is suggested. Our patients have been in a stable phase of the disease for more than one year since the given abnormalities were documented in their karyotypes. Further study is necessary to determine the prognostic significance of coexistence of these abnormalities in CLL patients.

Rearrangement of 14q32 in the Absence of t(14;18) Is Associated with Short Time to First Treatment in Chronic Lymphocytic Leukemia

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1438-1438
Author(s):  
Matthew S. Davids ◽  
Alexander R. Vartanov ◽  
Lillian Werner ◽  
Donna Neuberg ◽  
Paola Dal Cin ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Univariate Analysis ◽  
Multivariable Analysis ◽  
Lymphocytic Leukemia ◽  
Fish Analysis ◽  
Early Therapy ◽  
Cytogenetic Abnormalities ◽  
Increased Risk ◽  
Trisomy 12 ◽  
Time To First Treatment

Abstract Abstract 1438 The hierarchical cytogenetic classification for chronic lymphocytic leukemia (CLL) provides a valuable prognostic framework validated in several studies; however, the relationship between these cytogenetic abnormalities and IGHV and ZAP-70 has not been as well studied. Less is also known about the prognostic significance of different 14q32 rearrangements and combined cytogenetic abnormalities. We retrospectively examined cytogenetic abnormalities in 576 CLL patients seen at the Dana-Farber Cancer Institute between 2005 and the present. Interphase FISH analysis was performed using commercial probe sets (Abbott Molecular, Des Plaines, IL, USA) including P53/ATM, 12cen/13q14.1, and CCND1-IGH to exclude mantle cell lymphoma and identify chromosome 14q32 rearrangements, which were further evaluated with a t(14;18) probe. t(14;18) was evaluated separately from other 14q32 rearrangements. Fisher's exact test was used to assess the association of ZAP-70 and IGHV mutation status with cytogenetics. Median time to first treatment (TTFT) was summarized using Kaplan Meier estimates. In univariate analysis, the log rank test was used to assess associations of TTFT and cytogenetics, and in multivariable analysis, a Cox regression model was used. P-values were not adjusted for multiplicity of testing. The median age at diagnosis was 55 (range 25–86). At time of analysis, 43% (n=247) of patients had been treated, and 78% (n=452) of patients were still alive. Of 432 patients tested, 57.4% (n=248) had mutated IGHV, and of 420 patients tested, 42% (n=243) were ZAP-70 positive. Cytogenetic abnormalities of the 576 patients are summarized in table 1. Patients with 2 abnormalities were classified as the poorer risk abnormality by the Dohner hierarchy. The del(13q) group was significantly associated with negative ZAP-70 status (p = 0.003), whereas del(11q) was associated with positive ZAP-70 (p=0.02). A significant association was observed between unmutated IGHV and del(17p), del(11q), and trisomy 12 (p < 0.001 for each), and with complex cytogenetics (p = 0.03), whereas del(13q) had a significant association with mutated IGHV (p < 0.0001). In univariate analysis, a shorter median TTFT was associated with del(17p) vs. other (44 vs. 77 months, p = 0.0006), del(11q) vs. other (22 vs. 77 months, p < 0.0001), and 14q32 rearrangement vs. other (44 vs. 76 months, p=0.001), whereas a longer TTFT was observed for del(13q) vs. other (131 vs. 51 months, p < 0.0001). Of note, TTFT was not associated with trisomy 12 (p=0.20), trisomy 12 + del(13q) (p=0.61), IgH t(14;18) (p=0.57), normal cytogenetics (p=0.27), or complex cytogenetics (p=0.15). TTFT was similar between patients with mono-allelic, bi-allelic, or mixed del(13q) (131, 103, or 153 months, respectively, p = 0.97). In multivariable analysis, adjusting for ZAP-70 and IGHV, shorter TTFT was independently associated with 14q32 rearrangement (HR 2.51, 95% CI (1.41, 4.45), p=0.002) and del(17p) (HR 1.53, 95% CI (1.0, 2.33), p = 0.047). Del(13q) remained independently associated with longer TTFT (HR 1.97, 95% CI (1.40, 2.77), p < 0.0001). Overall, we show that high risk abnormalities such as del(17p) and del(11q) are associated with unmutated IGHV status and shorter TTFT, and that low risk del(13q) is associated with mutated IGHV, negative ZAP-70 status, and longer TTFT. Also, TTFT is similar between mono- and biallelic del(13q). Interestingly, trisomy 12 was highly associated with unmutated IGHV, but not shorter TTFT, suggesting that trisomy 12 may somehow mitigate the risk of unmutated IGHV. In contrast, 14q32 rearrangement was not associated with either IGHV or ZAP-70, but did predict a short TTFT of 44 months. This suggests that 14q32 rearrangement may hasten the need for treatment, regardless of IGHV or ZAP-70 status. Notably, 14q32 rearrangement due to t(14;18) did not confer increased risk of requiring early therapy. Ongoing analysis will assess overall survival for these cytogenetic abnormalities.Table 1.Cytogenetic abnormalities in 576 CLL patientsCytogenetic Abnormalityn%TTFT (months)del(13q)–total27548131· del(13q x 1)–mono-allelic18667.6% of del(13q)131· del(13q x 0)–bi-allelic238.4% of del(13q)103· del(13q)–mixed5118.5% of del(13q)153Normal701268Trisomy 12631154Trisomy 12 + del(13q)19353Del(17p)47844Del(11q)4172214q32 rearrangement25444IgH t(14;18)19395Complex (3 or more abnormalities)17358 Disclosures: No relevant conflicts of interest to declare.

Clinical Significance of 13q14 Number of Deleted Cells in Chronic Lymphocytic Leukemia

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4581-4581
Author(s):  
Giovanni Del Poeta ◽  
Francesca Maria Rossi ◽  
Maria Ilaria Del Principe ◽  
Michele Dal Bo ◽  
Annalisa Biagi ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Conflicts Of Interest ◽  
Normal Karyotype ◽  
Progression Free Survival ◽  
Intermediate Stage ◽  
Lymphocytic Leukemia ◽  
Prognostic Indicators ◽  
Fish Analysis ◽  
Clinical Difference ◽  
Trisomy 12

Abstract Abstract 4581 Cytogenetic aberrations are considered major prognostic indicators for predicting the survival of chronic lymphocytic leukemia (CLL) patients (Dohner et al, 2000) and interphase fluorescent in situ hybridization (I-FISH) with specific probes is generally used to detect the most frequent abnormalities. Moreover, deletion 13q14.3 on FISH analysis which is the most common cytogenetic abnormality in CLL is a favorable prognostic biomarker when detected as a sole abnormality, even if a higher percentage of 13q- nuclei was found to be associated with significantly shorter time to treatment (P<0.001), (van Dike et al, 2010; Dal Bo et al, 2011). Therefore, the primary endpoints of our research were: 1) to determine progression free survival (PFS) and overall survival (OS) on the basis of percentages of 13q- nuclei and 2) to confirm 13q14 number of deleted cells as an independent prognostic factor. We investigated 503 pts, median age 65 years (range 33–89), 291 males and 212 females. With regard to modified Rai stages at diagnosis, 163 had a low stage, 320 an intermediate stage and 20 a high stage. Probes for chromosome 13q (LSI-D13S319), 11q (LSI-ATM), 17p (LSI-p53) and chromosome 12 (CEP12) were used on nuclei collected at diagnosis. One hundred fifty-three patients (30.4%) exhibit a normal karyotype, 203 pts (40.4%) showed an isolated 13q-, 63 pts (12.5%) presented trisomy 12, 49 pts (9.7%) 11q deletion, 26 (5.2%) 17p deletion and 9 (1.8%) other chromosome abnormalities. Clearly, patients with intermediate/poor cytogenetic abnormalities (trisomy 12, del11q-, del17p-) showed significant shorter PFS and OS (7% vs 36% at 14 years and 45% vs 77% at 14 years, P<0.0001) in comparison with normal or del13q- pts. Maximally selected log-rank statistics identified the 50% of nuclei bearing del13S319 as the most appropriate cut-off value capable of separating del13q- cases into two subgroups with different PFS and OS distributions. In fact, pts with isolated 13q- in <50% of nuclei (110 pts) showed a longer PFS and OS (62% vs 16% at 12 years, P<0.0001 and 95% vs 47% at 16 years, P=0.0007, Figure) compared to those with ≥50% of nuclei (93 pts). Noteworthy, 64 (69%) of 93 of 13q- >50% pts had received chemotherapy at the time of analysis, whereas only 27 (25%) of 110 of 13q- <50% pts had been treated (P <0.0001). There was no significant clinical difference between heterozygous and homozygous 13q- patients as well as between 13q- cases with RB1 deletion (delRB1) and 13q- without delRB1. There was a significant correlation between number of 13q deleted nuclei and number of B-lymphocytes/microliter (P<0.0001) at diagnosis as well as between 13q- nuclei percentages and lymphocyte doubling time (P=0.001), meaning that 13q- nuclei represent both the amount of disease and the proliferation rate in CLL. Only slight significant correlations were found between 13q- percentages and CD38 (P=0.04) or ZAP-70 (P=0.01) or IgHV status (P=0.03), whereas 36 of 54 (67%) CD69+ pts had del13q- >50% (P=0.0003). In the context of del13q- subset, multivariate analysis of PFS confirmed the percentage of nuclei (P=0.00007) together with IgHV status (P=0.003) and ZAP-70 (P=0.0002) as an independent prognosticator. With regard to OS, percentage of nuclei (P=0.02) together with age (P=0.009) and IgHV status (P=0.03) was again confirmed as an independent variable. Therefore, the percentage of nuclei exhibiting 13q- at diagnosis has to be considered an important predictor of the clinical outcome and the clinical implications of an isolated 13q deletion in CLL appear more complex and important than originally considerated. Disclosures: No relevant conflicts of interest to declare.

THE ROLE OF THE GENETIC ABNORMALITIES, EPIGENETIC AND microRNA IN THE PROGNOSIS OF CHRONIC LYMPHOCYTIC LEUKEMIA

2018 ◽  
Vol 40 (4) ◽  
pp. 261-267 ◽  
Author(s):  
K Tari ◽  
Z Shamsi ◽  
H Reza Ghafari ◽  
A Atashi ◽  
M Shahjahani ◽  
...  
Keyword(s):  
B Cells ◽  
Chronic Lymphocytic Leukemia ◽  
Kinase Inhibitors ◽  
Bone Marrow Involvement ◽  
P53 Mutation ◽  
Lymphocytic Leukemia ◽  
Gene Promoters ◽  
Epigenetic Changes ◽  
Genetic Changes ◽  
Trisomy 12

Chronic lymphocytic leukemia (CLL) is increased proliferation of B-cells with peripheral blood and bone marrow involvement, which is usually observed in older people. Genetic mutations, epigenetic changes and miRs play a role in CLL pathogenesis. Del 11q, del l17q, del 6q, trisomy 12, p53 and IgVH mutations are the most important genetic changes in CLL. Deletion of miR-15a and miR-16a can increase bcl2 gene expression, miR-29 and miR-181 deletions decrease the expression of TCL1, and miR-146a deletion prevents tumor metastasis. Epigenetic changes such as hypo- and hypermethylation, ubiquitination, hypo- and hyperacetylation of gene promoters involved in CLL pathogenesis can also play a role in CLL. Expression of CD38 and ZAP70, presence or absence of mutation in IgVH and P53 mutation are among the factors involved in CLL prognosis. Use of monoclonal antibodies against surface markers of B-cells like anti-CD20 as well as tyrosine kinase inhibitors are the most important therapeutic approaches for CLL.

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