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Angiopoietin-2 expression in B-cell chronic lymphocytic leukemia: association with clinical outcome and immunoglobulin heavy-chain mutational status

Leukemia volume 21, pages 1312–1315 (2007)Cite this article

For several years chronic lymphocytic leukemia (CLL) has been considered a disease of accumulation of relentless mature-looking malignant monoclonal B cells due to a presumed defect in programmed cell death. However, several observations suggest that CLL cells are not immortal and that they require signals from the microenvironment to maintain viability. Increasing evidence shows that neovascularization plays a role in the biology of CLL.1 Increased microvessel density in marrow and lymph node and increased levels of certain pro-angiogenetic factors in the blood and urine of CLL patients were observed.2 Ang-2 has a pivotal role in the disruption of stability and quiescence of mature vasculature that coincides with the reinitiation of vascular remodelling and angiogenic sprouting in adult, as occurs in tumors. Abnormal serum levels of angiopoietin-2 (Ang-2) were found in several hematological malignancies.3, 4 The report of Hüttmann et al.,5 recently published in Leukemia, described an increase of Ang-2 plasma levels in the ZAP-70+CD38+ as compared to the ZAP-70−CD38− CLL patients. Moreover, Ang-2 plasma levels detected in Binet B and C patients exceeded those of Binet stage A cases, suggesting a correlation between Ang-2 secretion and CLL progression. Exploratory experiments of gene expression profiling (GEP) were previously performed in our laboratory on 29 unpurified CLL samples (% CD5+CD19+cells, 82±9%) (data deposited in NCBIs Gene Expression Omnibus http://www.ncbi.nlm.nih.gov/geo/, Accession:GSE4207) with Agilent Human-1A-Oligo-Microarray comprising 20 173 (60-mer) oligonucleotide probes using the protocol described elsewhere.6 Among the differentially expressed genes between 14 Ig-mutated CLL (M-CLL) and 15 Ig-unmutated CLL (UM-CLL) subsets, Ang-2 was found as one of the most up-regulated genes in UM-CLL relative to M-CLL (P=3.02 × 10−6, ANOVA). These results prompted us to undertake a survey of our own CLL patients to examine further the relationship between Ang-2 expression, immunoglobulin (Ig) mutational status and other established prognostic markers as well as progression-free survival.

We included 63 CLL patients (40 men, 23 women) in this study on the basis of the availability of peripheral blood (PB) at diagnosis with a high percentage of CD5+CD19+ cells (82±9%). The median age was 62 (range, 32–87) and the mean follow-up time was 3.8 years. Informed consent was provided according to the Declaration of Helsinki. IgVH gene sequence analysis was performed using the protocol of Marasca et al.7 Peripheral blood mononuclear cells (PBMCs) were obtained from B-CLL patients by density gradient centrifugation. In 10 CLL cases and in 9 normal controls, the CD19+ fraction was collected from peripheral blood using CD19Microbeads (Miltenyi Biotec, Auburn, CA, USA), obtaining a purity over 99%. Moreover, Ang-2 mRNA levels were measured by quantitative real-time PCR using SYBR Green reagents and an ABI Prism 7700 Sequence Detector (Applied Biosystems, Foster City, CA, USA), whereas Ang-2 protein expression was evaluated in the bone marrow (BM) of 10 CLL cases by immunohistochemistry using specific goat polyclonal antibodies (sc-7016; Santa Cruz Biotechnology, Inc., USA). Progression-free survival functions have been estimated using the product-limit (Kaplan–Meier) method and the curves have been compared between groups using log-rank test.

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References

  1. Peterson L, Kini AR . Angiogenesis is increased in B-cell chronic lymphocytic leukemia. Blood 2001; 97: 2529.

    Article  CAS  PubMed  Google Scholar 

  2. Kini AR, Kay NE, Peterson LC . Increased bone marrow angiogenesis in B cell chronic lymphocytic leukemia. Leukemia 2000; 14: 1414–1418.

    Article  CAS  PubMed  Google Scholar 

  3. Quartarone E, Alonci A, Allegra A, Bellomo G, Calabro L, D’Angelo A et al. Differential levels of soluble angiopoietin-2 and Tie-2 in patients with haematological malignancies. Eur J Haematol 2006; 77: 480–485.

    Article  CAS  PubMed  Google Scholar 

  4. Uneda S, Matsuno F, Sonoki T, Tniguchi I, Kawano F, Hata H . Expressions of vascular endothelial growth factor and angiopoietin-2 in myeloma cells. Haematologica 2003; 88: 113–115.

    CAS  PubMed  Google Scholar 

  5. Huttmann A, Klein-Hitpass L, Thomale J, Deenen R, Carpinteiro A, Nuckel H et al. Gene expression signatures separate B-cell chronic lymphocytic leukaemia prognostic subgroups defined by ZAP-70 and CD38 expression status. Leukemia 2006; 20: 1774–1782.

    Article  CAS  PubMed  Google Scholar 

  6. Marasca R, Maffei R, Zucchini P, Castelli I, Saviola A, Martinelli S et al. Gene expression profiling of acute promyelocytic leukaemia identifies two subtypes mainly associated with flt3 mutational status. Leukemia 2006; 20: 103–114.

    Article  CAS  PubMed  Google Scholar 

  7. Marasca R, Maffei R, Morselli M, Zucchini P, Castelli I, Martinelli S et al. Immunoglobulin Mutational Status Detected through Single-Round Amplification of Partial VH Region Represents a Good Prognostic Marker for Clinical Outcome in Chronic Lymphocytic Leukemia. J Mol Diagn 2005; 7: 566–574.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Author notes

  1. R Maffei and R Marasca: Both authors contributed equally to this work.

Authors and Affiliations

  1. Department of Oncology and Hematology, Hematology Unit, University of Modena and Reggio E, Modena, Italy

    R Maffei, R Marasca, S Martinelli, I Castelli, R Santachiara, P Zucchini, M Fontana, F Giacobbi, G Bonacorsi, P Temperani & G Torelli

  2. Excellence Environmental Carcinogenesis, Environmental Protection and Health Prevention Agency Emilia Romagna Region, Bologna, Italy

    E Morandi, P Silingardi & A M Colacci

  3. Hematology Unit, Arcispedale S. Maria Nuova, Reggio Emilia, Italy

    L Masini

  4. Department of Social, Cognitive and Quantitative Sciences, University of Modena and Reggio E, Reggio Emilia, Italy

    R Serra

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  1. R Maffei
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Correspondence to R Marasca.

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Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu)

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Maffei, R., Marasca, R., Martinelli, S. et al. Angiopoietin-2 expression in B-cell chronic lymphocytic leukemia: association with clinical outcome and immunoglobulin heavy-chain mutational status. Leukemia 21, 1312–1315 (2007). https://doi.org/10.1038/sj.leu.2404650

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