Strikingly Homologous Immunoglobulin Gene Rearrangements and Poor Outcome in VH3-21-Utilizing Chronic Lymphocytic Leukemia Independent of Geographical Origin and Mutational Status.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 175-175
Author(s):  
Fiona Murray ◽  
Mia Thorselius ◽  
Alexander Krober ◽  
Ulf Thunberg ◽  
Gerard Tobin ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Light Chain ◽  
Geographical Origin ◽  
Lymphocytic Leukemia ◽  
Gene Rearrangements ◽  
Immunoglobulin Gene ◽  
Mutation Status ◽  
Mutational Status ◽  
Significant Difference ◽  
Vh Genes

Abstract We recently reported that Swedish VH3-21-utilizing chronic lymphocytic leukemia (CLL) patients showed restricted immunoglobulin gene features and poor prognosis despite VH mutation status. To investigate whether VH3-21+ CLLs have similar characteristics in different parts of the world, we analyzed the VH and VL gene rearrangements in 90 patients from Sweden, Germany, Italy, USA, Finland and Australia and correlated these data with survival. Sixty-three percent of cases exhibited mutated VH genes and 37% had unmutated VH genes. Fifty patients (56%) displayed a short and homologous heavy-chain CDR3, many of these with the amino acid motif, DANGMDV. Also, a highly biased Vλ2-14 usage was evident in 73% of patients with a restricted light-chain CDR3, QVWDS(S/G)SDHPWV. Combined restricted heavy- and light-chain CDR3s were found in patients from all included countries. Although VH3-21+ CLLs have a remarkably predominant λ-expression, analyses of kappa deleting element showed a conserved rearrangement order of the light-chain loci. The overall survival was poor in the VH3-21+ cohort (median survival 88 months) with no significant difference in relation to mutation status or homologous/non-homologous CDR3. In summary, highly restricted B-cell receptors and worse outcome characterize VH3-21+ CLLs independent of geographical origin and mutation status. VH3-21 usage should now be included in prognostic stratification of CLL when assessing mutation status.

scholarly journals Mutation Status and Immunoglobulin Gene Rearrangements in Patients from Northwest and Central Region of Spain with Chronic Lymphocytic Leukemia

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
I. González-Gascón y Marín ◽  
J. A. Hernández ◽  
A. Martín ◽  
M. Alcoceba ◽  
M. E. Sarasquete ◽  
...  
Keyword(s):  
Overall Survival ◽  
Chronic Lymphocytic Leukemia ◽  
Central Region ◽  
Lymphocytic Leukemia ◽  
Gene Rearrangements ◽  
Immunoglobulin Gene ◽  
Mutation Status ◽  
Almost All ◽  
Time To First Treatment ◽  
Immunoglobulin Gene Rearrangements

The aim of this study was to investigate the frequency and mutation status of the immunoglobulin heavy variable chain (IGHV) in a cohort of 224 patients from northwest and central region of Spain diagnosed with chronic lymphocytic leukemia (CLL), and to correlate it with cytogenetic abnormalities, overall survival (OS) and time to first treatment (TTFT). 125 patients had mutated IGHV, while 99 had unmutated IGHV. The most frequently used IGHV family was IGHV3, followed by IGHV1 and IGHV4. The regions IGHV3-30, IGHV1-69, IGHV3-23, and IGHV4-34 were the most commonly used. Only 3.1% of the patients belonged to the subfamily IGHV3-21 and we failed to demonstrate a worse clinical outcome in this subgroup. The IGHV4 family appeared more frequently with mutated pattern, similar to IGHV3-23 and IGHV3-74. By contrast, IGHV1-69 was expressed at a higher frequency in unmutated CLL patients. All the cases from IGHV3-11 and almost all from IGHV5-51 subfamily belonged to the group of unmutated CLL.

KDE Analysis in Chronic Lymphocytic Leukemia Patients with B Cells Expressing IGHV3-21 and Discordant Mutational Status between Heavy and Light Chain.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1127-1127
Author(s):  
Emanuela M. Ghia ◽  
Laura Z. Rassenti ◽  
George F. Widhopf ◽  
Gregg J. Silverman ◽  
Andrew W. Greaves ◽  
...  
Keyword(s):  
B Cells ◽  
Chronic Lymphocytic Leukemia ◽  
Light Chain ◽  
Sequence Homology ◽  
Lymphocytic Leukemia ◽  
Light Chains ◽  
Receptor Editing ◽  
Mutation Status ◽  
Aggressive Disease ◽  
Mutational Status

Abstract Patients with chronic lymphocytic leukemia (CLL) cells that use IGHV3-21 typically have aggressive disease similar to that of patients with CLL cells that use unmutated IGHV. However, the IGHV3-21 genes expressed in CLL often have less than 98% sequence homology with the known germline IGHV3-21 and hence are considered mutated. However, many “mutated” IGHV3-21 in CLL have higher homology with the germline IGHV3-21 (e.g. >97%) than do most other IGHV in CLL that also are considered mutated (mean percent homology = 93 ± 4% (SD), N= 1,260). As such, some argue that 97% be used as the threshold for defining IGHV mutation status. To test whether this definition is valid for IGHV3-21 we examined the mutation status of the rearranged and/or expressed Ig light chain V genes. We examined 63 (2.6%) IgVH3-21 cases from a large cohort (N=2,457) of CLL patients evaluated by the CRC. Forty of 63 (63.5%) IGHV3-21 cases used ? and 23 (36.5%) used ? light chains. While the ?-expressing cases had perfect correlation in mutational status between IgH and IgL, 12 of 40 (30%) ?-expressing cases were discordant in that they had IGHV3-21 with 94–97.6% germline sequence homology and IGLV that were all unmutated. Nine of these 12 (75%) also displayed a strikingly homologous and short IgH third complementary determining region (CDR) motif (DANGMDV) and light chains encoded by IGLV3-21. To study the differentiation history of such cases, we examined for productively-rearranged, and formerly-expressed, IGKV alleles and non-functional IGKV alleles that had undergone rearrangement with the kappa deleting element (KDE). Analysis for IGKV-KDE rearrangements was performed on 7 of the 9 IGHV3-21/IGLV3-21 cases. Two cases had two non-functional IGKV alleles and five (71%) had one non-functional IGKV allele and one productively-rearranged IGKV allele that apparently was aborted via receptor editing. All 5 productively rearranged IGKV alleles had somatic mutations with germline-homology ranging from only 93.6% to 97.6% (median 96.3%). Non-conservative mutations were clustered in the CDRs, arguing that they were selected in an apparent antigen-driven response. All 9 non-functional IGKV alleles had 100% homology with known germline IGKV genes. For comparison, we examined for IGKV-KDE rearrangements in 4 cases of ?-expressing CLL cells that used the unmutated IGHV1-69 gene and IGLV3-09. All such cases each had two non-functional IGKV alleles that had 100% homology with known IGKV rearranged with KDE, indicating that none of these cases experienced IGKV receptor editing, a significantly lower incidence than that noted for cases using IGHV3-21/IGLV3-21 (P<0.01). These results indicate that despite sharing association with more aggressive disease, most CLL cases that use IGHV3-21/IGLV3-21 are distinctive from cases that use unmutated IGHV1-69 in their B cell differentiation history in that they are derived from B cells that formerly expressed ? light chains that had undergone somatic mutation and selection in an antigen-driven immune response prior to undergoing IGKV receptor editing. Despite expressing IGHV with >97% homology to the known germline IGHV3-21 and IGLV with >98% homology to known germline IGLV, these cases have experienced somatic mutation and apparent germinal center maturation prior to leukemogenesis.

scholarly journals Total Proteome Analysis Identifies Migration Defects As a Major Pathogenetic Factor in IGHV-Unmutated Chronic Lymphocytic Leukemia

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 718-718 ◽  
Author(s):  
Gina L Eagle ◽  
Rosalind E Jenkins ◽  
Kathleen J Till ◽  
Jithesh Puthen ◽  
Ke Lin ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Computational Analysis ◽  
Lymphocytic Leukemia ◽  
Differentially Expressed ◽  
Differentially Expressed Proteins ◽  
Data Set ◽  
Clinical Behaviour ◽  
Mutational Status ◽  
Significant Difference ◽  
Total Proteome

Abstract The mutational status of the immunoglobulin heavy chain variable region (IGHV) defines two clinically distinct forms of chronic lymphocytic leukemia (CLL) known as mutated (M-CLL) and un-mutated (UM-CLL). Patients with M-CLL usually have a favourable outcome whereas those with UM-CLL develop progressive disease and have shorter survival. However, the molecular mechanisms responsible for the more aggressive clinical behaviour associated with UM-CLL are not well understood. Here we describe the application of isobaric tags for relative and absolute quantification (iTRAQ) based mass spectrometry (MS) to analyse the total proteome of M-CLL and UM-CLL samples. This has enabled us to generate the largest quantity of proteomic information for CLL to date and, in particular, to directly compare the functions of differentially expressed proteins between UM-CLL and M-CLL cells through a systems biology approach. We isolated CLL cells from the peripheral blood from 18 CLL patients (9 UM-CLL, 9 M-CLL) and prepared cellular protein extracts which were digested and subjected to labelling with iTRAQ reagents, as previously described (Kitteringham et al, J Proteomics, 2010;73(8):1612-1631). Principal component analysis was used to assess variance across the data set generated by iTRAQ-MS. Statistical significance of the difference in the levels of expression of proteins between UM-CLL and M-CLL samples was determined using student T-test (2-tailed). Several differentially expressed proteins identified by iTRAQ-MS were also validated by immunoblotting. Computational analysis was performed to examine the functions of the differentially expressed proteins and their associated signalling pathways using the GeneGo pathway maps in the Metacore™ database (Thomson Reuters, NY, USA). Unsupervised clustering, based on the expression of 3521 identified proteins, separated CLL samples into two groups corresponding to IGHV mutational status. We identified 274 proteins that were differentially expressed between UM-CLL and M-CLL subgroups (p<0.05, Figure 1A). Hierarchical clustering based on the relative expression of differentially expressed proteins also separated individual CLL cases into two distinct clusters according to their IGHV status (Figure 1B). Computational analysis showed that 43 cell migration/adhesion pathways were significantly enriched (p<0.05) by 39 differentially expressed proteins, 35 of which were expressed at significantly lower levels in UM-CLL samples. Furthermore, UM-CLL cells under-expressed proteins associated with cytoskeletal remodelling and over-expressed proteins associated with transcriptional and translational activity. Taken together, these findings indicated that UM-CLL cells are less migratory and more adhesive than M-CLL cells, resulting in their retention in lymph nodes where they are exposed to proliferative stimuli. In agreement with this hypothesis, analysis of an extended cohort of 120 CLL patients revealed that twice as many patients with UM-CLL than M-CLL had documented lymphadenopathy (50% v 24%; P<0.01). The association between UM-CLL and lymphadenopathy was not simply a reflection of increased tumour burden as there was no significant difference in the leukocyte count between the two groups (medians of 37 x 109/L and 28 x 109/L, respectively; P>0.05). In addition, other pathways that promote cell survival and proliferation in UM-CLL cells were also enriched by the differentially expressed proteins. These include the immune response pathway involving B-cell receptor (BCR) signalling (P=0.006), the endoplasmic reticulum (ER) stress response pathway (P=0.035) and the Wnt signalling pathway (P=0.006). Our study has shown that quantitative analysis of the total proteome by iTRAQ-MS was able to separate individual CLL cases according to IGHV status and explained the more aggressive clinical behaviour of UM-CLL and its particular sensitivity to novel therapeutic agents that induce anatomical displacement from the lymph node microenvironment, such as ibrutinib and idelalisib. Moreover, in keeping with the ability of proteomics to detect alterations in gene expression resulting from both transcriptional and post-transcriptional mechanisms, the study illustrates the considerable potential of iTRAQ-MS coupled with computational analysis to elucidate pathogenetic mechanisms and indicate therapeutic strategies in cancer. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

Distinctive IgVH Gene Segments Usage and Mutation Status in Chinese Patients with Chronic Lymphocytic Leukemia.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4708-4708
Author(s):  
Lijuan Chen ◽  
Yaping Zhang ◽  
Wenjuuan Zheng ◽  
Jianyong Li ◽  
Changgeng Ruan
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Gene Family ◽  
Gene Families ◽  
Lymphocytic Leukemia ◽  
Chinese Patients ◽  
Chain Gene ◽  
Mutation Status ◽  
Significant Difference ◽  
Variable Heavy ◽  
Vh Gene

Abstract Chronic lymphocytic leukemia (CLL) is characterized by the relentless accumulation of monoclonal B cells with the appearance of small mature lymphocytes and a characteristic CD5 and CD19 co-expression immunophenotype. The incidence of CLL in Asian countries is lower than that in the Western ones, where CLL is the most common leukemia. To evaluate the frequency and mutation status of immunoglobulin (Ig) variable heavy chain gene (IgVH) expression in Chinese patients with CLL. We investigated IgVH gene segments usage and mutation status by multiplex RT-PCR in 52 CLL patients, and analyzed the relationship between IgVH somatic mutation status and the expression of CD38, ZAP-70 and CLLU1. 38 patients had mutated IgVH, and 14 had unmutated IgVH. The most frequently expressed VH gene family was found to be VH3 (46.2%) followed by VH4 (40.4%), VH1 (5.8%), VH2 (5.8%) and VH7 (1.9%), with no expression of VH5 and VH6 gene families. VH1-69 and VH3-21 which commonly overused in Western CLL weren’t detected in our cohort. The frequency of IgVH gene families indicates significant difference in Chinese CLL patients compared with Western patients, suggesting involvement of ethnic and/or environmental factors in CLL disease initiation. IgVH gene mutation status was significantly associated with the expression of CD38 and CLLU1. The expression of them may be simple and reliable surrogates for the identification of IgVH mutations. VH gene family usage and mutation status VH family n Mutated VH gene Unmutated VH gene VH1 3 3 0 VH2 3 2 1 VH3 24 19 5 VH4 21 16 5 VH5 0 0 0 VH6 0 0 0 VH7 1 0 1

scholarly journals 98% IGHV Gene Identity Is Still the Optimal Cutoff to Predict the Prognosis of Chronic Lymphocytic Leukemia Patients in China

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 5545-5545
Author(s):  
Yi Xia ◽  
Ke Shi ◽  
Qian Sun ◽  
Chun Qiao ◽  
Huayuan Zhu ◽  
...  
Keyword(s):  
Prognostic Factor ◽  
Chronic Lymphocytic Leukemia ◽  
Conflicts Of Interest ◽  
Somatic Hypermutation ◽  
Lymphocytic Leukemia ◽  
Study Cohort ◽  
Optimal Cutoff ◽  
Entire Cohort ◽  
Mutational Status ◽  
Significant Difference

Abstract Background: Immunoglobulin heavy chain variable region (IGHV) has been an important prognostic factor for chronic lymphocytic leukemia (CLL) for decades. 98% being a cut-off value for IGHV is a mathematical choice and researches on the best cut-off value have never been stopped. Chinese CLL patients are known to differ from Caucasian CLL patients on both clinical and genetical features. However, the optimal cutoff for IGHV mutational status has not yet been studied in this particular ethnic group. Method: We carried out a study on 595 Chinese CLL patients in order to find out whether 98% is the best cut-off value for IGHV in Chinese CLL patients. Genomic DNA from peripheral blood or bone marrow was subjected to PCR amplification following the IGH Somatic Hypermutation Assay v2.0 protocol (InVivoScribe). Sequences were aligned to ImMunoGeneTics/V-QUEry and Standardization (IMGT/-VQUEST) database. Result: 600 sequences were received after IGHV rearrangement sequencing. IGHV3-23, IGHV4-34, IGHV3-7, IGHV4-39 and IGHV1-69 were the most frequently used IGHV genes. 352 (58.7%) cases were IGHV-mutated while 248 (41.3%) cases were IGHV-unmutated if the classical 98% classification by ERIC was used. In order to determine the optimal cut-off value, we used 1% as the interval to divide the entire cohort into 7 groups according to the mutational rate, which were <95%, 95%-95.99%, 96%-96.99%, 97%-97.99%, 98%-98.99%, 99%-99.99% and 100% respectively. Binet A patients had a relatively indolent course of disease and cases with different IGHV mutational rates had no significant differences in time to first treatment (TTFT) apart from truly unmutated (100%) cases. For the whole study cohort, significant difference appeared at 98% interval (P<0.001 and P=0.005 for TTFT and OS respectively) while intervals less than 98% had no significant difference compared with the <95% group. Similarly, there was no clear dissimilarities among 98%, 99% and 100% intervals (Table 1a and b). All the other prognostic factors including del(17p), del(11q), TP53 mutation, MYD88 mutation, NOTCH1 mutation, SF3B1 mutation, CD38, ZAP-70, Binet staging, gender, β2-microglobulin and EBV-DNA were differently distributed between group <98% and group ³98%, but not among subgroups in ³98%. In multivariate analysis, the 98% IGHV was also an independent prognostic factor for TTFT and OS. Conclusion: 98% is the optimal cutoff value for IGHV mutational status to predict the prognosis of CLL patients in China. Table 1. Table 1. Disclosures No relevant conflicts of interest to declare.

Demonstration of Biclonal Chronic Lymphocytic Leukemia with Mutated and Unmutated Clones and Concurrent but Clonally Unrelated Myeloma in the Same Patient by Single Cell Immunoglobulin Heavy and Light Chain Gene Analysis.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4772-4772
Author(s):  
Selim A. Yavuz ◽  
Dan-Paul Hartmann ◽  
Said Baidas ◽  
Peter E. Lipsky ◽  
Metin Ozdemirli
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
B Cell ◽  
Single Cell ◽  
Light Chain ◽  
Lymphocytic Leukemia ◽  
Chain Gene ◽  
Gene Rearrangements ◽  
Kappa Light Chain ◽  
B Cell Malignancies ◽  
Myeloma Cells

Abstract Patients with chronic lymphocytic leukemia (CLL) may develop other B cell malignancies in their clinical course including aggressive diffuse large B-cell lymphomas and rarely myelomas. In a large proportion of cases, the secondary B cell malignancies reflected the emergence of immunophenotypically and genetically different clones. An immature type plasma cell myeloma developed in a 73-year-old female patient in whom CLL was diagnosed four years previously. The CLL expressed CD5, CD19, CD23, CD38 and surface kappa light chain, but were negative for ZAP-70. Trisomy 12 was detected by FISH analysis. PCR analysis of the peripheral blood for immunoglobulin heavy chain genes demonstrated two sharp bands that were initially interpreted as biallelic heavy chain gene rearrangements. Myeloma cells were CD38 and CD138 positive, CD19 negative and expressed cytoplasmic kappa light chain, but not heavy chains. In order to investigate the clonal relationship between these B cell malignancies, a detailed analysis of VHDJH and VκJκ gene rearrangements in individually sorted CD5 and CD19 double-positive CLL cells and also in CD38-positive and CD19-negative myeloma cells by single cell PCR of genomic DNA and direct sequencing was carried out. This technique permitted identification and pairing of both the heavy and light chain immunoglobulin genes from the same individually sorted cell. A total of 17 individual CLL and 23 myeloma cells were successfully analyzed. Our analysis demonstrated (a) the presence of two discrete clones of CLL, one with usage of [VH1-2*04/D3-3*01/J3*02]-[Vκ2-28*01/J1*01] without VH and Vκ hypermutation and the other with usage of [VH1-2*04/D4-11*01/J6*02]-[Vκ1-5*03/J1*01] with VH and Vκhypermutation; (b) no clonal relationship between the CLL and myeloma cells that utilized different VHDJH and VκJκ rearrangements [VH3-66*02/3-10*01/J4*03]-[Vκ1-33*01/J2*02] with VH and Vκ hypermutation. To our knowledge, this is the first demonstration of a biclonal CLL with mutated and unmutated clones in the same patient along with a third clonally unrelated B cell malignancy. This result suggests that single cell analysis may be necessary to detect subtle biclonality of CLL that might be associated with a more aggressive phenotype.

Relative Value of CD38 and ZAP-70 Versus Immunoglobulin Mutation Status in Predicting Early Disease Progression in Chronic Lymphocytic Leukemia.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2778-2778 ◽  
Author(s):  
Laura Z. Rassenti ◽  
Donna S. Neuberg ◽  
Lang Huynh ◽  
William G. Wierda ◽  
John G. Gribben ◽  
...  
Keyword(s):  
Chronic Lymphocytic Leukemia ◽  
Variable Region ◽  
Lymphocytic Leukemia ◽  
Added Value ◽  
Logrank Test ◽  
Region Gene ◽  
Mutation Status ◽  
Mutational Status ◽  
Relative Value ◽  
Median Level

Abstract We assessed the relative value of CD38 in predicting the need for early treatment in 307 patients with chronic lymphocytic leukemia (CLL) previously characterized for ZAP-70 and immunoglobulin heavy-chain-variable region gene (IgVH) mutational status (NEJM2004, 351;9;893–901). Recursive partitioning by maximally selected chi-square analyses of flow cytometry and clinical data identified the optimal cut-off for designating a CLL sample as CD38+ was at 34%, which was highly similar to the conventional cut-off of 30%. Since these cut-offs identified highly similar cohorts of CD38+ CLL patients (e.g. 32.2% (99/307) vs. 35.5% (109/307)) we chose to use 30% as the cut-off for this analysis. We used the logrank test to compare CD38 expression-status to the time from diagnosis to therapy (TTT), initiated per established NCI-Working Group guidelines. Patients with CLL cells that were CD38-negative by this criterion had a median TTT of 7.8 years; whereas, patients with CD38+ CLL cells had a median TTT of 3.7 years (p<0.0001). There was a significant association between expression of CD38 and ZAP-70 or use of unmutated IgVH genes (p < 0.0001). The median level of CD38 among cases that used mutated IgVH was 4.0%, compared to 27.4% among cases that used unmutated IgVH (p < 0.0001). The median level of CD38 among ZAP-70-negative cases was 5.3% (range from 0.1% to 99.5%), compared to 27.2% among ZAP-70-positive cases (range from 0.1% to 99.4%) (p < 0.0001). We explored the TTT based only on flow cytometric parameters (CD38 and ZAP-70) and investigated whether addition of mutation status significantly altered our predictions. For most patients, the knowledge of CD38 and ZAP-70 provided a reliable means for predicting the TTT and the knowledge of the IgVH mutation status did not substantially alter the prediction. Only in 41 cases (14% of the total), where the CLL cells were CD38+ but negative for ZAP-70, was the prediction significantly improved by inclusion of IgVH mutation status. The median TTT of this group of 41 patients was 7.8 years. Addition of mutation status identified a subgroup of 20 patients with unmutated IgVH genes for whom median TTT was estimated at 4.6 years; 21 patients with mutated IgVH genes had a median TTT of 8.4 years. For patients with CLL cells that were CD38-negative and either negative or positive ZAP-70, stratification via IgVH mutation status did not identify subgroups within each of these categories that had significantly different median TTT. We conclude that knowledge of CD38 and ZAP-70, as assessed by the CRC, can reliably predict TTT in most patients with CLL. For the 14% of cases that are CD38+ but negative for ZAP-70, determining the IgVH mutation status may provide added value in predicting the time from diagnosis to initial therapy.

Sign in / Sign up

Export Citation Format

Share Document