CD38 expression is a poor predictor for VH gene mutational status and prognosis in chronic lymphocytic leukemia

Abstract The mutational status of tumor immunoglobulin VHgenes is providing a powerful prognostic marker for chronic lymphocytic leukemia (CLL), with patients having tumors expressing unmutated VH genes being in a less favorable subset. However, the biologic differences correlating with VH gene status that could determine the clinical course of the disease are unknown. Here we show that differing responses to IgM ligation are closely associated with VH gene status. Specifically, 80% of cases with unmutated VH genes showed increased global tyrosine phosphorylation following IgM ligation, whereas only 20% of samples with mutated VH genes responded (P = .0002). There was also an association between response to IgM ligation and expression of CD38 (P = .015). The Syk kinase, critical for transducing B-cell receptor (BCR)– derived signals, was constitutively present in all CLL samples, and there was a perfect association between global phosphorylation and induction of phosphorylation/activation of Syk. Nonresponsiveness to anti-IgM could be circumvented by ligation of IgD (10 of 15 samples tested) or the BCR-associated molecule CD79α (12 of 15 samples tested). These results suggest that multiple mechanisms underlie nonresponsiveness to anti-IgM in CLL and that retained responsiveness to anti-IgM contributes to the poor prognosis associated with the unmutated subset of CLL. The prognostic power of the in vitro response to IgM ligation remains to be determined in a large series, but the simple technology involved may present an alternative or additional test for predicting clinical course.

Download Full-text

Vegf and bFGF Single-Nucleotide Polymorphisms In Chronic Lymphocytic Leukemia: Impact On Susceptibility

Blood ◽

10.1182/blood.v122.21.5287.5287 ◽

2013 ◽

Vol 122 (21) ◽

pp. 5287-5287

Author(s):

Sandra Ballester ◽

Begoña Pineda ◽

Eduardo Tormo ◽

Blanca Navarro ◽

Ariadna Perez ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Conflicts Of Interest ◽

Expression Patterns ◽

Lymphocytic Leukemia ◽

P Value ◽

Nucleotide Polymorphisms ◽

Exact Test ◽

Cd38 Expression ◽

Mutational Status ◽

The Individual

Abstract Background B-cell chronic lymphocytic leukemia (B-CLL) is a heterogeneous disease with a highly variable clinical outcome. Recent studies have identified a number of different molecular prognostic markers (including mutational status of the IgVH gene, ZAP70 and CD38 expression) that allow to discriminate patients in prognostic subgroups. However, different expression patterns of angiogenic factors as VEGF, VEGFR1 and bFGF have been related with B-CLL susceptibility and treatment requirements. We have analyzed the polymorphisms: -710 C/T in VEGFR1, rs1109324, rs1547651, rs3025039 (936C/T) and rs833052 in VEGF and rs1449683 (223 C/T) in bFGF in order to determine the possible association with susceptibility in B-CLL. Methods Peripheral blood samples from 230 B-CLL patients and 476 healthy controls were genotyped using probes TaqMan SNP Genotyping Assays. Samples were providing from the Hospital Clinic of Valencia. Four SNPs in the VEGF gene, one SNP in the bFGF gene and one SNP in the VEGFR1 gene were evaluated. Statistical analysis was performed using SNPStats program (Catalan Institute of Oncology) and Fisher's exact test was applied to evaluate the significance. Results We have observed an increased frequency of the T allele in the rs1449683 SNP [OR 1.62 (95% CI: 0.98-2.66) p-value =0.063] and in the rs1547651 SNP [OR 0.72 (95% CI: 0.51-1.03), p-value=0.072] in our B-LLC patients when compared to control subjects. Moreover we observed that T allele carriers of rs3025039 (VEGF) have a significant protective effect concerning this disease [OR 0.59 (95% CI: 0.39-0.89) p-value=0.009]. Conclusion Our data indicate an increased frequency of the T allele in polymorphisms rs1449683 (bFGF) and rs1547651 (VEGF) in the group of patients, which possibly account for the individual susceptibility to develop B-CLL. On the other hand the data provided suggest that the T allele of VEGF rs3025039 is likely important genetic marker of susceptibility to B-CLL. Further studies regarding the role of pro-angiogenic markers in B-CLL would be beneficial to help elucidate pathogenic pathways in this disease. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

Comprehensive Assessment of Genetic and Molecular Features Predicting Outcome in Patients With Chronic Lymphocytic Leukemia: Results From the US Intergroup Phase III Trial E2997

Journal of Clinical Oncology ◽

10.1200/jco.2006.08.3089 ◽

2007 ◽

Vol 25 (7) ◽

pp. 799-804 ◽

Cited By ~ 251

Author(s):

Michael R. Grever ◽

David M. Lucas ◽

Gordon W. Dewald ◽

Donna S. Neuberg ◽

John C. Reed ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Prognostic Significance ◽

Variable Region ◽

Hazard Ratio ◽

Lymphocytic Leukemia ◽

Phase Iii ◽

Therapeutic Trial ◽

Cd38 Expression ◽

Molecular Features ◽

Mutational Status

Purpose Genomic features including unmutated immunoglobulin variable region heavy chain (IgVH) genes, del(11q22.3), del(17p13.1), and p53 mutations have been reported to predict the clinical course and overall survival of patients with chronic lymphocytic leukemia (CLL). In addition, ZAP-70 and Bcl-2 family proteins have been explored as predictors of outcome. Patients and Methods We prospectively evaluated the prognostic significance of a comprehensive panel of laboratory factors on both response and progression-free survival (PFS) using samples and data from 235 patients enrolled onto a therapeutic trial. Patients received either fludarabine (FL; n = 113) or fludarabine plus cyclophosphamide (FC; n = 122) as part of a US Intergroup randomized trial for previously untreated CLL patients. Results Complete response (CR) rates were 24.6% for patients receiving FC and 5.3% for patients receiving FL (P = .00004). PFS was statistically significantly longer in patients receiving FC (median, 33.5 months for patients receiving FC and 19.9 months for patients receiving FL; P < .0001). The occurrence of del(17p13.1) (hazard ratio, 3.428; P = .0002) or del(11q22.3) (hazard ratio, 1.904; P = .006) was associated with reduced PFS. CR and overall response rates were not significantly different based on cytogenetics, IgVH mutational status, CD38 expression, or p53 mutational status. Expression of ZAP-70, Bcl-2, Bax, Mcl-1, XIAP, Caspase-3, and Traf-1 was not associated with either clinical response or PFS. Conclusion These results support the use of interphase cytogenetic analysis, but not IgVH, CD38 expression, or ZAP-70 status, to predict outcome of FL-based chemotherapy. Patients with high-risk cytogenetic features should be considered for alternative therapies.

Download Full-text

CD38 expression and Ig VH gene mutation in B-cell chronic lymphocytic leukemia

Blood ◽

10.1182/blood.v97.6.1902 ◽

2001 ◽

Vol 97 (6) ◽

pp. 1902-1902 ◽

Cited By ~ 62

Author(s):

Zoltan Matrai ◽

Ke Lin ◽

Michael Dennis ◽

Paul Sherrington ◽

Mirko Zuzel ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

B Cell ◽

Gene Mutation ◽

Lymphocytic Leukemia ◽

Cd38 Expression ◽

Vh Gene ◽

Cell Chronic Lymphocytic Leukemia

Download Full-text

Biological Prognostic Markers in Chronic Lymphocytic Leukemia

Acta Medica (Hradec Kralove Czech Republic) ◽

10.14712/18059694.2016.99 ◽

2009 ◽

Vol 52 (1) ◽

pp. 3-8 ◽

Cited By ~ 8

Author(s):

Vladimíra Vroblová ◽

Lukáš Smolej ◽

Filip Vrbacký ◽

Karolína Jankovičová ◽

Monika Hrudková ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Clinical Course ◽

Variable Region ◽

Lymphocytic Leukemia ◽

Initial Assessment ◽

Western World ◽

Cd38 Expression ◽

Cytogenetic Aberrations ◽

Mutational Status

Chronic lymphocytic leukemia (CLL) is the most frequent leukemic disease of adults in the Western world. It is remarkable by an extraordinary heterogeneity of clinical course with overall survival ranging from several months to more than 15 years. Classical staging sytems by Rai and Binet, while readily available and useful for initial assessment of prognosis, are not able to determine individual patient’s ongoing clinical course of CLL at the time of diagnosis, especially in early stages. Therefore, newer biological prognostic parameters are currently being clinically evaluated. Mutational status of variable region of immunoglobulin heavy chain genes (IgVH), cytogenetic aberrations, and both intracellular ZAP- 70 and surface CD38 expression are recognized as parameters with established prognostic value. Molecules regulating the process of angiogenesis are also considered as promising markers. The purpose of this review is to summarize in detail the specific role of these prognostic factors in chronic lymphocytic leukemia.

Download Full-text

Correlative Study of ZAP-70 Gene Expression by Quantitative PCR and CD-38 Surface Marker in Patients with Chronic Lymphocytic Leukemia (CLL).

Blood ◽

10.1182/blood.v104.11.4796.4796 ◽

2004 ◽

Vol 104 (11) ◽

pp. 4796-4796

Author(s):

Melissa L. Larson ◽

Jamile M. Shammo ◽

Sari H. Enschede ◽

Parameswaran Venugopal ◽

Daniel Russell ◽

...

Keyword(s):

Flow Cytometry ◽

Prognostic Factors ◽

Chronic Lymphocytic Leukemia ◽

Median Number ◽

Lymphocytic Leukemia ◽

Stage I ◽

Cd38 Expression ◽

Mutational Status ◽

Multiple Treatments

Abstract Patients with chronic lymphocytic leukemia have heterogeneous clinical courses. Many possible prognostic factors have been evaluated as a way to predict each patient’s future course. Two such prognostic factors are IgVH gene mutational status and CD38 expression. Recently, ZAP-70 expression has been found to correlate with IgVH gene mutation, but little is known about the correlation between ZAP-70 and CD 38 expression. We sought to evaluate the concordance between the two. METHODS: A procedure was developed at our institution to evaluate ZAP-70 transcripts by PCR. The results were then confirmed by flow cytometry. The blood or bone marrow samples of 14 patients diagnosed with CLL were analyzed by PCR for ZAP-70 transcripts and flow cytometry for ZAP-70 protein expression and CD38 expression. The results of the ZAP-70 transcripts and CD38 expression were then correlated to the clinical courses of the patients. RESULTS: Of the 14 patients analyzed, the majority had Rai Stage I disease at diagnosis (4 Stage 0, 8 Stage I, and 2 Stage II). There was a median follow-up of 28 months. The median number of treatments for these patients was one (range 0–6). Overall, the results for ZAP-70 and CD38 expression were concordant in eight with CD38 data unavailable for 2 patients. In the four patients with discordant results, 3 of them had high CD38 expression and low ZAP-70. However, patients with high ZAP-70 were more likely to require multiple treatments once treatment was indicated. Patients with high CD38 were also more likely to be treated. Cytogenetic data was available for 9 patients with the following results: 5 with normal cytogenetics, 1 with deletion of 11q, 1 with deletion of 13q, and 2 with deletion of 17p. Five of the nine patients had high ZAP-70 expression. CONCLUSIONS: In our institution, the results of ZAP-70 and CD38 expression were concordant 67% of the time. Longer follow-up is necessary and may further delineate the roles of cytogenetics, ZAP-70 expression, and CD38 expression on prognosis in CLL.

Download Full-text

Elevated Levels of IL-17, MIP-1 β, MIG and ITAC in B-Cell Chronic Lymphocytic Leukemia: Correlation with Prognostic Markers.

Blood ◽

10.1182/blood.v108.11.2788.2788 ◽

2006 ◽

Vol 108 (11) ◽

pp. 2788-2788

Author(s):

Xiao J. Yan ◽

Michael Centola ◽

Igor Dozmorov ◽

Rajendra N. Damle ◽

Steven L. Allen ◽

...

Keyword(s):

B Cells ◽

Chronic Lymphocytic Leukemia ◽

B Cell ◽

Stromal Cells ◽

Lymphocytic Leukemia ◽

Serum Cytokines ◽

Cd38 Expression ◽

Cytokines And Chemokines ◽

Mutational Status ◽

Cell Chronic Lymphocytic Leukemia

Abstract B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the progressive accumulation of mature B cells. The growth and accumulation of B-CLL cells requires survival and migratory signals from endogenously produced cytokines and chemokines, many of which are supplied by stromal cells. In this comparative study we analyzed the expression, as a function of B-CLL disease, of a set of inflammatory and immune cytokines and chemokines known to regulate the growth, survival and/or trafficking of B cells. Serum cytokines were measured in 87 B-CLL patients and in 32 healthy subjects using a combination of multiplex cytokine bead and protein array technologies. A subset of cytokines and chemokines were found to be significantly elevated in serum from B-CLL patients as compared to healthy age-matched controls: IL-17 (p<0.01), MIP-1β (p<0.001), MIP-1α (p<0.01), IL-6 (p<0.05), IL-8 (p<0.001), IL-10 (p<0.001), IL-12 (p<0.05), MIG (p<0.01), and ITAC (p<0.001). Serum levels of IL-1β, IL-1α, IL-2, IL-4, IL-5, IL-12, IL-13, IL-15, TNFα, IFNγ, IFNα, GM-CSF, Eotaxin, and MCP-1 were not significantly different in B-CLL patients as compared to healthy age-matched controls. Previous studies have reported elevated levels of IL-6, IL-8, IL-10, and MIG in B-CLL, and these molecules may contribute anti-apoptotic and growth-promoting signals that allow the expansion of B-CLL cells. IL-17 is known to promote angiogenesis and induce stromal cell expression of cytokines and chemokines that support B-CLL survival, suggesting potential mechanisms whereby this cytokine may impact on B-CLL biology. MIG and ITAC are ligands for CXCR3, a chemokine receptor expressed on B-CLL cells. These chemokines positively affect the responsiveness of pDC to the homing chemokine, SDF-1, raising the possibility that their elevation in B-CLL may lead to increased responsiveness of B-CLL cells to SDF-1, a stromal factor that enhances their survival. MIP-1α and MIP-1β are inflammatory chemokines that regulate cell recruitment and activation. Interestingly, MIP-1α and MIP-1β trigger stromal cells to produce RANKL, which has recently been suggested to contribute to B-CLL pathogenesis by triggering production of IL-8. B-CLL patients divide into populations that present with (a) stable disease and favorable prognostics including mutated VH and low CD38 expression, and (b) progressive disease with poor prognostics including unmutated VH and high CD38. When serum cytokines were analyzed as a function of select prognostic indicators discrete differences were observed. IL-8 levels correlated with mutated VH, IL-17 and IL-6 levels correlated with unmutated VH, and MIG, IP-10 and ITAC levels correlated with high CD38 expression. MIP-1α and MIP-1β levels differed significantly from controls irrespective of mutational status or CD38 expression. The correlation of IL-17, which in addition to its stromal activating properties is known to promote angiogenesis, with unmutated VH suggests a potential role for this cytokine in mediating the increased angiogenic phenotype that has been observed in poor prognosis CLL.

Download Full-text

Mcl-1 expression has in vitro and in vivo significance in chronic lymphocytic leukemia and is associated with other poor prognostic markers

Blood ◽

10.1182/blood-2008-05-157131 ◽

2008 ◽

Vol 112 (9) ◽

pp. 3807-3817 ◽

Cited By ~ 156

Author(s):

Chris Pepper ◽

Thet Thet Lin ◽

Guy Pratt ◽

Saman Hewamana ◽

Paul Brennan ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Gene Mutation ◽

Prognostic Markers ◽

Critical Role ◽

Lymphocytic Leukemia ◽

Mutation Status ◽

Cd38 Expression ◽

Vh Gene

Bcl-2 family proteins play a critical role in the regulation of apoptosis in chronic lymphocytic leukemia (CLL). However, their association with established prognostic markers is unknown. In this study, we analyzed the expression of Bcl-2, Bax, and Mcl-1 in 185 CLL patients and evaluated their relationship with other prognostic markers, in vitro sensitivity to fludarabine, and clinical outcome. Mcl-1 expression was significantly correlated with stage of disease (P < .001), lymphocyte doubling time (P = .01), VH gene mutation status (P < .001), CD38 expression (P < .001), and ZAP-70 expression (P = .003). In addition, Mcl-1 and Mcl-1/Bax ratios showed strong correlations with in vitro resistance to fludarabine (P = .005 and P < .001, respectively). Furthermore, elevated Mcl-1 expression and Mcl-1/Bax ratios were predictive of time to first treatment in the whole cohort (P < .001 and P < .001, respectively) and in stage A patients only (P = .002 and P = .001, respectively). Taken together, our data show that Mcl-1 is a key controller of in vitro drug resistance and is an important regulator of disease progression and outcome in CLL. It therefore represents a promising therapeutic target in this incurable condition. The close correlation between Mcl-1 expression and VH gene mutation status, CD38 expression, and ZAP-70 expression offers a biologic explanation for their association with adverse prognosis.

Download Full-text

ZAP-70 Expression Is More Predictive Than VH Gene Mutational Status of BCR-Mediated Tyrosine Phosphorylation, NF-κB Activation and CLL Cell Survival.

Blood ◽

10.1182/blood.v106.11.2942.2942 ◽

2005 ◽

Vol 106 (11) ◽

pp. 2942-2942

Author(s):

Chris Pepper ◽

Suhair Alghazal ◽

Paul Brennan ◽

Clare Rowntree ◽

Guy Pratt ◽

...

Keyword(s):

B Cell ◽

Cell Survival ◽

Tyrosine Phosphorylation ◽

Surrogate Marker ◽

Cell Receptor ◽

B Cell Receptor ◽

Lymphocytic Leukemia ◽

Cd38 Expression ◽

Mutational Status ◽

Vh Gene

Abstract We analysed tyrosine phosphorylation, NF-κB activation and survival in a cohort of 64 chronic lymphocytic leukemia patients. By graphing VH gene mutational status, CD38 expression and ZAP-70 expression as continuous variables, we investigated correlations between these prognostic markers and cell signaling. There was a strong correlation between CD38 expression and basal levels of tyrosine phosphorylation (r2 = 0.68). VH gene mutational status and ZAP-70 expression correlated with the induction of tyrosine phosphorylation by anti-IgM (r2 = 0.31 and 0.46 respectively). These findings suggest that tyrosine phosphorylation integrates these three prognostic phenotypes. In the context of NF-κB signaling, the percentage change in tyrosine phosphorylation (post anti-IgM) and ZAP-70 expression correlated with NF-κB activation (r2 = 0.2 and 0.19 respectively) and the ability to activate NF-κB was strongly correlated with cell survival (r2 = 0.67). The therapeutic rationale for the inhibition of NF-κB was strengthened by our observations that the NF-κB inhibitor, BAY 11–7082, caused a concentration-dependent increase in apoptosis in CLL samples. Taken together, our data suggest that CD38 is a surrogate marker of basal cellular activation in CLL that is independent of the B-cell receptor. Furthermore, ZAP-70 expression is the most important predictor of tyrosine phosphorylation, downstream NF-κB activation and CLL cell survival following B-cell receptor cross-linking. These findings provide a mechanism-based rationale for the poor prognosis of CD38+ and ZAP-70+ CLL patients.

Download Full-text

The pattern of CD38 expression defines a distinct subset of chronic lymphocytic leukemia (CLL) patients at risk of disease progression

Blood ◽

10.1182/blood-2002-06-1801 ◽

2003 ◽

Vol 101 (4) ◽

pp. 1262-1269 ◽

Cited By ~ 177

Author(s):

Paolo Ghia ◽

Giuseppe Guida ◽

Stefania Stella ◽

Daniela Gottardi ◽

Massimo Geuna ◽

...

Keyword(s):

Chronic Lymphocytic Leukemia ◽

Lymphocytic Leukemia ◽

Progression Rate ◽

Cd38 Expression ◽

Mutational Status ◽

Patients At Risk ◽

Leukemic Clone ◽

Patient Groups ◽

Risk Of Disease ◽

Igvh Mutational Status

Chronic lymphocytic leukemia (CLL) has a variable clinical course. CD38 expression and IgVH gene mutational status are independent predictors of prognosis, but their relationships and the CD38 cutoff level are unknown. Using cytofluorography, we analyzed CD38 in 148 patients, in 108 of whom we were able to evaluate IgVH mutations, make correlations with disease history, and assess cumulative survival. Three different patient groups were identified by the CD38 expression pattern: a group homogeneously CD38−, a group homogeneously CD38+, and a group characterized by a bimodal profile, because of the concomitant presence of variable proportions of 2 distinct populations, one CD38+ and one CD38−. In CD38 bimodal expression patients the CD38+ subset was significantly more represented in the bone marrow than in the peripheral blood. For IgVH mutations, 11.4% of CD38−, 84.6% of CD38+, and 68.0% of CD38 bimodal expression patients had no mutation. CD38 expression, IgVH mutational status, and traditional prognostic factors were concordant. The progression rate was 12.9% for CD38−, 75.0% for CD38+, and 63.3% for CD38 bimodal expression patients. Only 25.8% of the CD38−patients but 63.3% of the bimodal and 75.0% of CD38+patients were treated. The presence of a CD38+ population, albeit small, correlated with the development of autoimmune manifestations. The CD38− group has not yet reached the median survival, which is 183 months in the CD38+ group and 156 months in the CD38 bimodal expression group, regardless of the size of the CD38+ population. The presence of a distinct CD38+ population within the leukemic clone, rather than a numerical cutoff definition, correlates with IgVH gene mutational status and, irrespective of its size, identifies CLL patients who will have progressive disease.

Download Full-text