DETECTION of CHROMOSOMAL 14q32 ABNORMALITIES IN B-CELL CHRONIC LYMPHOCYTIC LEUKEMIA.

Abstract Abstract 4385 Introduction Conventional cytogenetic and FISH studies with the standard panel (13q14, 11q22.3, 17p13, 12, 6q23) are used to detect chromosomal abnormalities of clinical significance in patients with chronic lymphocytic leukemia (B-CLL). Recently, translocations and 5'IGH deletions involving chromosome14q32 are associated with B-CLL. Aim Our aim was to investigate the presence and the characteristics of chromosome 14q32 aberrations in a group of patients with B-CLL. Patients and Methods A total of 58 patients with B-CLL were investigated by conventional cytogenetic, in addition the cultures were stimulated with CpG oligodeoxynucleotide plus IL2. FISH studies with 14q32/IGH break-apart probe designed to detect chromosomal breakage of the IGH (14q32) locus, were done for 59 patients. Results Chromosomal abnormalities were detected in 60.3% of cases by cytogenetic. 14 patients showed complex cariotype while trisomy 12 was the most frequent anomaly found in 8 patients. Among the twelve other patients several chromosomal aberrations were noted: del(13)(q14), del(13)(q12q21), del(13)(q12q14), del(13)(q13q32), +21, t(11;13)(q23;q14), +12 t(1;4)(q31;p14), t(3;14)(p21;q22), del (11)(q12) +21, del(6)(q21), inv3(?p13q21), del(11)(q12). Abnormalities of chromosome 14 were found in 23.8% of patients. Deletion of 5'IGH, corresponding to the variable IGH segment, was the most frequent anomaly found in 8 patients. Interesting, a 3'IGH deletion was detected in two patients, while only one patient showed a complete deletion of chromosome 14 (47,XXY,add14q32). Three patients showed 14q32 translocations involving the IGH locus. Conclusions Based on our findings, deletions of the variable region of the IGH gene (IGHv) and 14q32/IGH translocations are involved in B-CLL. As these preliminary data are based on small sample size, our goal will be to study the cytogenetic profile of a large number of CLL patients. Future studies will permit the identification of 14q32 translocations and the type and the frequency of IGH rearrangements. Finally, chromosome 14 abnormalities will be correlated to other cytogenetic and FISH abnormalities, and associations with known prognostic markers, such as IGVH mutation status and ZAP-70 expression, will be investigated. Disclosures: No relevant conflicts of interest to declare.

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Hodgkin lymphoma arising in patients with chronic lymphocytic leukemia: outcomes from a large multi-center collaboration

Haematologica ◽

10.3324/haematol.2020.256388 ◽

2020 ◽

pp. 0-0

Author(s):

Deborah M. Stephens ◽

Ken Boucher ◽

Elizabeth Kander ◽

Sameer A. Parikh ◽

Erin M. Parry ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Hodgkin Lymphoma ◽

Hematopoietic Cell Transplantation ◽

Small Sample Size ◽

Case Series ◽

Standard Of Care ◽

Median Number ◽

Small Sample ◽

Lymphocytic Leukemia ◽

Treatment Type

Chronic lymphocytic leukemia (CLL) patients who develop Hodgkin lymphoma (HL) have limited survival. No current therapeutic standard of care exists. We conducted a multi-center retrospective study of patients with Hodgkin Transformation (HT) of CLL. Clinicobiologic characteristics, treatment type, and survival outcomes were analyzed and compared with historic case series. Ninety-four patients were identified. Median age at HT was 67 years (range, 38-85). Median time from CLL diagnosis to HT was 5.5 years (range, 0-20.2). Prior to HT, patients received a median of 2 therapies for CLL (range, 0-12). As initial therapy for HT, 61% (n=62) received ABVD-based regimens (adriamycin, bleomycin, vinblastine, and dacarbazine). Seven (7%) patients received hematopoietic cell transplantation (HCT) while in first complete remission (CR1). The median number of treatments for HT per patient was 1 (range, 0-5) with 59 (61%) patients only receiving one line of therapy. After HT, patients had a median follow-up of 1.6 years (range, 0-15.1). Two-year overall survival (OS) after HT diagnosis was 72% (95%CI 62-83%). The patients who received standard ABVD-based therapy had a median OS of 13.2 years. Although limited by small sample size, the patients who underwent HCT for HT in CR1 had a similar 2-year OS (n=7; 67%) compared to patients who did not undergo HCT for HT in CR1 (n=87; 72%; p=0.46). In this multi-center study, HT patients treated with ABVD-based regimens had prolonged survival supporting the use of these regimens as standard of care for these patients.

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In Chronic Lymphocytic Leukemia Females Survive for Longer Than Males Even in Patients with Unmutated Immunoglobulin Genes.

Blood ◽

10.1182/blood.v104.11.16.16 ◽

2004 ◽

Vol 104 (11) ◽

pp. 16-16 ◽

Cited By ~ 2

Author(s):

Terry John Hamblin ◽

Jenny A. Orchard ◽

Zadie A. Davies ◽

Rachel E. Ibbotson ◽

Ann C. Gardiner ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Cell Biology ◽

Chromosomal Abnormalities ◽

Alkylating Agents ◽

Variable Region ◽

Lymphocytic Leukemia ◽

First Line ◽

Free Survival ◽

Significant Difference ◽

Males And Females

Abstract Among the proposed prognostic factors for chronic lymphocytic leukemia (CLL), gender has been constant. In published trials females survive longer. With the new prognostic markers that relate to cell biology it seemed that an explanation was forthcoming. The sex ratio for patients with unmutated immunoglobulin variable region heavy chain (IgVH) genes is M:F=2:1 whereas for those with mutated IgVH genes it is close to unity. Females are less likely to have the more malignant subset. We have explored whether this is a sufficient explanation for the better outcome in females. 310 patients with CLL were studied. 260 were local patients representing the referral pattern of a district hospital and 50 were second-opinion referrals. 180 were male and 130 female. Overall, the median survival of females was 161 months compared to 119 months for males (p<0.005). 189 (101 males; 88 females) had mutated and 121 (79 males; 42 females) unmutated IgVH genes. As expected patients with mutated IgVH genes had longer median survivals (mutated 183 months, unmutated 95 months; p<0.0001). Thus males are more likely to belong to the shorter-lived subtype. However, this is not a complete explanation. Among those with unmutated IgVH genes, females also survived significantly longer (135 months versus 97 months; p<0.05), whereas among those with mutated IgVH genes there was no difference. Censoring for unrelated deaths did not affect the findings. We also looked at treatment-free survival. This was better for females than males (median time to treatment not yet reached versus 113 months; p=0.0097) but among those with unmutated IgVH genes there was no significant difference (49 months versus 31 months; p=0.57). However, survival following first treatment was significantly better for females than for males both for the whole group (127 months versus 70 months; p=0.0092) and for the unmutated subgroup (100 months versus 45 months; p=0.014), but there was no difference for the mutated group. This finding is consistent with the findings in clinical trials for which survival is calculated from date of entry into the trial, not date of diagnosis. There were other slight differences between males and females. Mean age at presentation was slightly greater for females (66.7 v 64.3; p=0.0063), and females who were treated were more likely to have received alkylating agents alone (20 males, 17 females; p=0.0015) but neither of these factors is likely to have led to greater longevity. 117 patients (86 males; 41 females) had required treatment. Apart from the group treated with alkylating agents alone there were no significant differences in how males and females were treated. 44 patients received an alkylating agent followed by fludarabine (30 males and 14 females), and 27 received a fludarabine containing regimen as first line (22 males and 5 females). Seven patients received monoclonal antibodies (5 male, 2 females) and there were 3 allografts (two male, one female) and 1 autograft (male). The prevalence of adverse chromosomal abnormalities as detected by FISH was 6.4% for 17p13 deletions and 12.3% for 11q23 deletions and not significantly different (separately or together) between the sexes. We conclude that females survive for longer following treatment for CLL for unknown reasons. We are aware that androgen receptors play a part in lymphopoiesis, but how this relates to the gender differences in CLL is unclear.

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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 ◽

10.1182/blood.v108.11.2092.2092 ◽

2006 ◽

Vol 108 (11) ◽

pp. 2092-2092 ◽

Cited By ~ 4

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.

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Evidence for Autostimulatory Mechanisms in Chronic Lymphocytic Leukemia.

Blood ◽

10.1182/blood.v120.21.2880.2880 ◽

2012 ◽

Vol 120 (21) ◽

pp. 2880-2880

Author(s):

Martin Trepel ◽

Fabian Muller ◽

Mareike Frick ◽

Janina Rahlff ◽

Claudia Wehr ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Phage Display ◽

B Cell ◽

Conflicts Of Interest ◽

Specific Binding ◽

Variable Region ◽

Cell Receptor ◽

B Cell Receptor ◽

Lymphocytic Leukemia ◽

Fab Fragment

Abstract Abstract 2880 Background: The development and / or course of chronic lymphocytic leukemia (CLL) may be driven by the recognition of antigens through the B cell receptor (BCR). While it has been recognized that the diversity of epitope recognition may be astonishingly confined in CLL, knowledge on antigens recognized by CLL BCRs is still limited. Here, we identified and characterized an epitope recognized by a defined CLL BCR which may broaden our view on potential mechanisms of antigenic drive in CLL. Methods: The B- cell receptor of a random CLL-patient was cloned and expressed as Fab fragment in E.coli. Random phage display reptile litanies we skeletal on the immobilized Fab and landed peptides were tested for specific binding. Specific clones we sequenced and sequences were analyzed for homology to known proteins. Recognition of candidate proteins was verified in brooding assays or recombinant proteins. Results: Screening random phage display peptide libraries, we identified a CLL BCR epitope mimic that displayed a high degree of homology to a conserved peptide string in the variable region of immunoglobulin heavy and light chains. CLL BCR binding to this epitope as well as binding to full length heavy and light immunoglobulin chains was verified by binding assays and a protein array screening. Interestingly, the CLL BCR also interacted with itself, as the identified epitope was also present in its own primary amino acid sequence. Conclusions: These findings suggest the possibility of self-recognition of BCRs within the CLL cell membrane or BCR interactions between neighboring CLL cells. This may potentially result in autostimulation of the leukemic cell independent of “exogenous” antigens and may account for self-sufficient signaling of some CLL-BCRs in driving disease progression. As the peptide mimicking this immunoglobulin epitope is known to be recognized by BCRs of other CLL cases in addition to the index case investigated here, such autostimulatory mechanisms may be relevant to a large number of CLL patients. Disclosures: No relevant conflicts of interest to declare.

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TP53-Induced Glycolysis and Apoptosis Regulator Protects from Spontaneous Apoptosis and Predicts Poor Prognosis in Chronic Lymphocytic Leukemia

Blood ◽

10.1182/blood.v128.22.5101.5101 ◽

2016 ◽

Vol 128 (22) ◽

pp. 5101-5101

Author(s):

Ming Hong ◽

Yi Xia ◽

Yu Zhu ◽

Huihui Zhao ◽

Yue Xie ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Conflicts Of Interest ◽

Aerobic Glycolysis ◽

Variable Region ◽

Lymphocytic Leukemia ◽

Newly Diagnosed ◽

Spontaneous Apoptosis ◽

Free Survival ◽

Stromal Microenvironment ◽

Binet Stage

Abstract The circulating chronic lymphocytic leukemia (CLL) cells appear not to be overly utilizing aerobic glycolysis. However, CLL cells' recurrent contact with the stromal microenvironment leads to an increase of aerobic glycolysis and the cells' overall glycolytic capacity, which promotes cell survival and proliferation. TP53-induced glycolysis and apoptosis regulator (TIGAR) has been directly implicated in cellular metabolism in the control of glycolysis. TIGAR inhibits glycolysis and protects cells from intracellular reactive oxygen species (ROS)-associated apoptosis. Here, we investigated correlation between TIGAR expression and apoptosis in CLL primary cells, along with its relationship with the clinical characteristics and prognosis in 102 newly diagnosed CLL patients. Our data showed TIGARoverexpression correlated with protection from spontaneous apoptosis in CLL cells, and is strongly associated with advanced Binet stage, unmutated immunoglobulin heavy-chain variable region (IGHV) status, CD38 positivity, β2-microglobulin and p53 aberrations. Higher expression of TIGAR was associated with briefer treatment-free survival (median: 3 months vs. 51 months, p = 0.0108) and worse overall survival (median: 74 months vs. not reached, p = 0.0242), as well as their diverse responses to fludarabine based chemotherapy. TIGAR expression of the patients who were resistant to chemotherapy was significantly higher than those who were sensitive to chemotherapy (mean: 0.3859±0.1710 vs. 0.0974±0.0291, p = 0.0290). Taken together, our findings depict how bioenergetics characteristics could be therapeutically exploited in CLL. Disclosures No relevant conflicts of interest to declare.

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Surface IgM of Chronic Lymphocytic Leukemia Cells Displays Unusual Glycans Indicative of Antigen Engagement In Vivo.

Blood ◽

10.1182/blood.v114.22.55.55 ◽

2009 ◽

Vol 114 (22) ◽

pp. 55-55

Author(s):

Graham Packham ◽

Serge Krysov ◽

Christopher Ian Mockridge ◽

Kathy N Potter ◽

Freda K Stevenson

Keyword(s):

B Cells ◽

Chronic Lymphocytic Leukemia ◽

Conflicts Of Interest ◽

Variable Region ◽

Immunoglobulin M ◽

Lymphocytic Leukemia ◽

Glycosylation Pattern ◽

Mature Form

Abstract Abstract 55 Several lines of evidence support the idea that surface immunoglobulin M (sIgM) plays a key role in determining the clinical behavior of chronic lymphocytic leukemia (CLL). For example, the presence of somatic mutations in immunoglobulin variable region genes is a strong prognostic marker with unmutated CLL (U-CLL) associated with a poor outcome relative to mutated CLL (M-CLL). U-CLL also generally express higher levels of sIgM and retain the ability to signal via this receptor. In this study, we used surface biotinylation to analyse sIgM in CLL and discovered that it exists in two forms with differing mobility on SDS-PAGE. Treatment with glycosidases revealed that these forms were due to different N-glycosylation patterns in the μ constant region. One form is similar to that of normal B cells in bearing mature complex glycans common to most cell surface glycoproteins. The other is an immature mannosylated form more characteristic of endoplasmic reticulum (ER)-located μ chains. CLL cells expressed variable proportions of the immature mannosylated form and quantitative analysis demonstrated that, on average, the proportion of mannosylated sIgM was approximately 2-fold higher (p=0.006) in U-CLL compared to M-CLL. Although normal B cells isolated from blood expressed only the mature form of sIgM, in vitro treatment with anti-μ resulted in upregulation of the immature form, suggesting that glycan modification is a consequence of antigen exposure. Consistent with this, in vitro incubation of CLL cells was associated with increased expression of the mature form of sIgM. Phosphotyrosine analysis demonstrated that both forms of sIgM were able to signal following sIgM engagement in vitro. Taken together, these findings support the concept that CLL cells are continuously exposed to antigen in vivo. This process leads to a change in the N-glycosylation pattern of the re-expressed sIgM so that a mannosylated form predominates, especially in U-CLL. Strikingly, expression of mannosylated sIgM is also characteristic of follicular lymphoma, where it is constitutively displayed via N-glycosylation sites in the Ig variable region (Radcliffe et al. J Biol Chem. 2007; 282, 7405-15). Persistent mannosylation of sIgM appears to be a feature common to several B-cell malignancies, suggesting a role in pathogenesis. Disclosures: No relevant conflicts of interest to declare.

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SNP-Arrays: a Routine Cytogenetic Tool In Chronic Lymphocytic Leukemia ?.

Blood ◽

10.1182/blood.v116.21.4611.4611 ◽

2010 ◽

Vol 116 (21) ◽

pp. 4611-4611

Author(s):

Alban Godon ◽

Franck Genevieve ◽

Malgorzata Truchan-Graczyk ◽

Laurence Baranger ◽

Virginie Eclache ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Chromosomal Abnormalities ◽

Conflicts Of Interest ◽

Snp Array ◽

Lymphocytic Leukemia ◽

Primary Data ◽

Assay Sensitivity ◽

Snp Arrays ◽

Median Size ◽

Trisomy 12

Abstract Abstract 4611 Background: Chronic lymphocytic leukemia (B-CLL) is the most common adult leukemia in Western countries, and is characterized by a highly variable clinical course. Interphase fluorescent in situ hybridization (I-FISH) has been able to identify chromosomal abnormalities in ~80% of B-CLL, including deletions at 13q, 11q, 17p and trisomy 12, which has proven to be prognostic indicators for disease progression and survival. Although recent immunostimulatory methods have substantially improved analysis via conventional metaphase cytogenetics (CC), detection of chromosome changes is limited by the low mitotic activity of CLL cells in vitro. High-density single nucleotide polymorphism (SNP) arrays are commercially available technologies, which allow genome-wide detection of allelic copy number gains or losses, and loss-of-heterozygosity (LOH) regions. Aims: To assess whether SNP-arrays are more sensitive than I-FISH and CC to detect specific chromosomal abnormalities associated with prognosis in B-CLL, i.e. del13q, del11q, del17p and tri12. Methods: Blood samples from 24 patients with B-CLL at diagnosis were tested in parallel by I-FISH, CC and SNP-array. FISH: blood smear samples were hybridized with 4 probes, in order to detect deletions at 17p13.1, 11q22.3, 13q14.3, and trisomy 12 [respectively LSI p53, ATM, D13S319 and centromeric CEP12 probes, Abbott]. In normal lymphocytes, an average of 6.7% nuclei showed one signal (truncated nuclei), and we defined the cut-off level for detection of a deletion at 11% (mean+3SD). The cut-off for detection of tri12 was defined at 5% (mean+3SD). CC: blood lymphocytes are cultivated for 72 hours with immunostimulants (DSP30 and IL-2) and metaphases analyzed according to standard procedures. SNP-arrays: DNA samples (200 ng) were hybridized on the Illumina HumanCNV370-quad v3 BeadChips, which assess 373,397 markers with a median marker spacing of 4.9 kb (mean 7.8 kb). The I-Scan system was used to scan the BeadChips (primary data). GenomeStudio 1010 v1 and CNVPartition 2.4.4 package were used to process primary data and identify chromosomal deletions/amplifications and LOH regions. Results: I-FISH identified deletions at 13q (15-95% of nuclei; mean=51%), 11q (35-54%; mean=43%) and trisomy 12 (32-49%; mean=37%) in 17 (71%) [including 2 cases of biallelic deletions], 3 (12.5%) and 4 (17%) cases, respectively. No del17p was detected. Five B-CLL cases presented associated FISH abnormalities. SNP-arrays identified all changes (100%) detected by I-FISH (del13q, median size: 13 Mb – range, 0.49–50 Mb; del11q, median size: 39 Mb – range, 34.8–42 Mb]. However, SNP-arrays showed 5 additional deletions at 13q14.3. Three patients had cryptic deletions (~52 to 82 kb), not detected by the FISH LSI D13S319 probe (~130 kb) or CC, and two others had large deletions (1.1 and 33.3 Mb) but with one signal below the cut-off of 11% by I-FISH (therefore considered as negative- both had normal CC). In these two cases, tumor cell enrichment before I-FISH allowed the detection of the deletion. In one case with del13q and tri12 (30% of nuclei by FISH), an additional del11q was also detected by SNP-array and CC (in 5/24 mitoses). The size of this deletion was 37.8 Mb and involved the cytogenetic bands 11q14.1 to 11q23.2 (including ATM). In addition, SNP-arrays enabled to define more precisely the size and location of the abnormalities. For instance, in one case with tri12 (as identified by the centromeric FISH probe), a partial trisomy of chromosome 12 short arm was indeed detected by SNP-array. No 17p abnormality was detected by SNP-array (either deletion or LOH). Only one of the 24 samples had no abnormality by SNP, I-FISH or CC for the loci studied. Conclusions: Despites using a relatively low density SNP-array (~370,000 markers), a higher sensitivity of Illumina BeadChips was observed. “SNP+/FISH negative” cases can be explained by either cryptic deletions or low leukemic cell content (< cut-off level). “SNP+/CC negative” cases can be explained by either a higher resolution, or the presence of normal mitosis in excess following stimulation. Higher density SNP-arrays (> 5 million markers) may be used to improve the assay sensitivity, especially regarding del13q14 which seems to be present in a great majority of our cases. Our study confirms usefulness of SNP-arrays for prognostic evaluation of B-CLL patients at diagnosis, and suggests the use of this assay as a routine procedure. Disclosures: No relevant conflicts of interest to declare.

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Karyotype-specific microRNA signature in chronic lymphocytic leukemia

Blood ◽

10.1182/blood-2009-06-229211 ◽

2009 ◽

Vol 114 (18) ◽

pp. 3872-3879 ◽

Cited By ~ 132

Author(s):

Rosa Visone ◽

Laura Z. Rassenti ◽

Angelo Veronese ◽

Cristian Taccioli ◽

Stefan Costinean ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Chromosomal Abnormalities ◽

Variable Region ◽

Normal Karyotype ◽

Lymphocytic Leukemia ◽

High Expression ◽

Mutation Status ◽

Ighv Gene ◽

Trisomy 12 ◽

Disease Initiation

Abstract Chromosomal abnormalities, immunoglobulin heavy chain variable–region (IGHV) gene mutation status, and ζ-associated protein 70 (ZAP-70) expression levels have independent prognostic relevance in chronic lymphocytic leukemia (CLL); however, their concordance is variable. Because deregulation of microRNAs has been linked to disease initiation and progression in CLL, we studied the value of the microRNAs as a signature for CLL patients with specific chromosomal abnormalities. We identified 32 microRNAs able to discriminate the 11q deletion, 17p deletion, trisomy 12, 13q deletion, and normal karyotype cytogenetic subgroups. The expression values of 9 among the 32 microRNAs (miR-151-3p, miR-34a, miR-29c, miR-29b, miR-155, miR-148a, miR-146a, miR-146b5p, and miR-640) were correlated with gene expression data from the same samples to assess their biologic impact on CLL. In this study we also found that IGHV unmutated, high expression of ZAP-70 protein, and low expression of the miR-223, miR-29c, miR-29b, and miR-181 family were strongly associated with disease progression in CLL cases harboring 17p deletion, whereas in those harboring trisomy 12 only high expression of the miR-181a, among the analyzed parameters, suggested more aggressive disease. Thus, the use of the microRNA-based classifications may yield clinically useful biomarkers of tumor behavior in CLL.

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Molecular Cytogenetics Analysis of 13q14 Biallelic Deletion in Chronic Lymphocytic Leukemia: A Study on 250 Patients

Blood ◽

10.1182/blood.v118.21.1454.1454 ◽

2011 ◽

Vol 118 (21) ◽

pp. 1454-1454

Author(s):

Alessandro Gozzetti ◽

Giulia Papini ◽

Rosaria Crupi ◽

Adele Frasconi ◽

Francesco Forconi ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Chromosomal Abnormality ◽

Chromosomal Abnormalities ◽

Conflicts Of Interest ◽

Strong Association ◽

Lymphocytic Leukemia ◽

The Other ◽

Median Percentage ◽

Mutational Status ◽

13Q Deletion

Abstract Abstract 1454 Deletion 13q14 (13q-) detected by fluorescence in situ hybridization (FISH) is the most frequent chromosomal abnormality in chronic lymphocytic leukemia (CLL). When 13q- is detected as sole abnormality has a good prognosis, while aggressive outcome is registered when 13q- is combined with other chromosomal abnormalities such as del 11q or del 17p. A recent study evidenced also that patients with higher percentage of 13q-deleted cells (>70%) are at higher risk for aggressive disease. Some studies evidenced that 13q- deletion size (involving D13S319 +/−Rb1) seems to matter in terms of time to treatment (TTT) and prognosis (OS). Few studies evaluated so far the incidence and prognosis of a biallelic 13q- deletion, i.e. the deletion of both alleles of the minimal deleted region (MDR) of chromosome 13q.In particular prognosis has been reported controversial. We analyzed at single institution 250 CLL patients by FISH in order to evaluate the incidence and prognosis of biallelic 13q- by using probes for D13S319 and RB1 that map to DLEU2/MIR15A/MIR16-1 and RB1 loci. Results were correlated in terms of TTT and OS with IGHV mutational status (mutated vs unmutated), RAI/BINET stage, CD38 positivity and/ or ZAP-70 positivity, beta-2M, LDH, other chromosomal abnormality (+12, del17p, del11q). Deletion 13q was considered present if >10% of nuclei were deleted out of 300 nuclei scored by two different and independent observers. All biallelic cases were confirmed by FISH using a probe for LSI-D13S319 and 13q34 to exclude false positive results. 135/250 (52%) patients presented a monoallelic del 13q. 45/135 (32%) presented a monoallelic del of RB1 while 20/135 (15%) cases presented a biallelic 13q-.12/20 (80%) presented a biallelic 13q- as sole abnormality, while 8/20 presented a 13q- associated with other cytogenetic abnormalities (one 17p-, five 11q-, two +12). The median percentage of 13q-deleted cells was 50% (range 14–86). Median age was 65 (range 50–85), M/F 12/8; 80% of the patients were RAI stage 0–1, while only 10% were RAI stage 4. No differences were seen in patients with biallelic deletion of 13q when LDH, b2M, ZAP-70,IGHV were considered. CD38 was negative in 16/20 patients. Regarding the MDR of chromosome 13q, 11/20 patients presented a biallelic del of D13S319, while 5/20 had a biallelic deletion of RB1; 5/20 patients presented a mono+biallelic del of D13S319 while 3/20 a mono+biallelic deletion of RB1. When we analyzed clinical and biological characteristics comparing patients with biallelic13q-,monoallelic 13q- and with no 13q-, we did not find differences in terms of: stage (RAI-Binet), P=0.2,P=0.9; B2 M, P=0.4; LDH,P=0.1; CD38 positivity, P=0.2; ZAP-70, P=0,1; IGHV M vs UM,P=0.65; P53 mutated vs wild type, P=0.1; del 11q was significantly associated more with biallelic 13q-, P=0.02. TTT and OS were not significantly different between biallelic 13q- patients and the other two groups (164 vs 212 vs 211, P=0.9). 8/20 patients with biallelic 13q- have been treated, all 5 patients carrying also a 11q- received treatment, the other 3 patients had: 1patient RB1 deletion in 92% of cells and 2 deletion 13q- in>75% of cells. In conclusion, biallelic 13q- are present in about 8% of all cases of CLL and in 15% of 13q- patients. A strong association with del 11q was found and this correlated with disease progression and treatment.CD38 was negative in the majority of patients with biallelic 13q-. RB1 was deleted in 32% of 13q- patients. No differences were found in terms of clinical characteristics, TTT and OS with monoallelic 13q-. Disclosures: No relevant conflicts of interest to declare.

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Chromosome Study On Chronic Lymphocytic Leukemia Using CpG- Oligodeoxynucleotide as Immunostimulant Agent.

Blood ◽

10.1182/blood.v114.22.4383.4383 ◽

2009 ◽

Vol 114 (22) ◽

pp. 4383-4383

Author(s):

Yafang Wu ◽

Yongquan Xue ◽

Li Yao ◽

Hui Jiang ◽

Jun Zhang ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Detection Rate ◽

Conflicts Of Interest ◽

Interleukin 2 ◽

Lymphocytic Leukemia ◽

Banding Technique ◽

Cpg Odn ◽

Partial Deletion ◽

Karyotypic Analysis ◽

Cpg Oligodeoxynucleotide

Abstract Abstract 4383 Objective To verify whether CpG-oligodeoxynucleotide (CpG-ODN) can raise the detection rate of the karyotypic abnormalities in chronic lymphocytic leukemia (CLL). Methods The bone marrow (BM) or peripheral blood (PB) cells from 57 cases of CLL were collected and cultured with CpG-ODN DSP30+interleukin-2 (IL-2), phytohemagglutinin (PHA), pokeweed(PWM) or IL-2, respectively. 5 days later cells were harvested for chromosome preparation. Karyotypic analysis was performed using R banding technique. Panel fluorescence in situ hybridization (FISH) was carried out on 19 cases of CLL with normal karyotype using following probes: Cen12, D13S25, Rb1, ATM, P53, MYB and IgH. Genomic DNA from 21 cases of them was extracted from BM or PB. The immunoglobulin variable heavy chain (IgVH) were amplified by polymerase reaction (PCR) and sequenced. CD38 and ZAP70 expressions in leukemic cells were determined by flow cytometry (FCM). Results The detection rate of karyotypic abnormalities in CpG-ODN+IL-2 group (43.85%) was obviously higher than PHA (15.09%), PWM (17.31%) and IL-2 (3.13%) groups (P<0.01). 52 karyotypic abnormalities were found. Among them, trisomy12 (+12) or +12 with other abnormalities were the most common, while translocations were the most frequent structural abnormalities including 3 unbalanced and 11 balanced translocations among which 7 had 14q32 rearrangements. 13 cases showed one or more abnormalities on FISH examination including trisomy12 and P53 deletion each in one case, IgH rearrangement and partial deletion each in one case, 13q14.3 deletion in 11 cases of which 5 cases also had Rb1 deletion, one case had Rb1 partial deletion. No cases with ATM or MYB deletions were found. IgVH mutation was detected in 10/21 cases by PCR and sequencing. FCM showed 10/45 cases expressed CD38, while 11/27 cases expressed ZAP70. Among 26 cases examined for CD38 and ZAP70 expressions simultaneously, 5 cases were CD38+ZAP70+, 13 cases CD38-ZAP70-, 6 cases CD38-ZAP70+, 2 cases CD38+ZAP70-, respectively. Statistics disclosed a correlation between complex karyotype and IgVH without mutation, but no association between karyotype and CD38 or ZAP70 expression. Conclusion CpG-ODN immunostimulation can obviously raise the detection rate of abnormal karyotypes, especially balanced and unbalance translocations in CLL. FISH is important complement to conventional karyotypic analysis, the combination of both methods can provide more comprehensive genetic information for CLL. Disclosures: No relevant conflicts of interest to declare.

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