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Bone marrow microvessel density and plasma angiogenic factors in myeloproliferative neoplasms: clinicopathological and molecular correlations

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Abstract

Increased angiogenesis in BCR-ABL1 negative myeloproliferative neoplasms (MPNs) has been recognized, but its connection with clinical and molecular markers needs to be defined. The aims of study were to (1) assess bone marrow (BM) angiogenesis measured by microvessel density (MVD) using CD34 and CD105 antibodies; (2) analyze correlation of MVD with plasma angiogenic factors including vascular endothelial growth factor, basic fibroblast growth factor, and interleukin-8; (3) examine the association of MVD with clinicopathological and molecular markers. We examined 90 de novo MPN patients (30 polycythemia vera (PV), primary myelofibrosis (PMF), essential thrombocythemia (ET)) and 10 age-matched controls. MVD was analyzed by immunohistochemistry “hot spot” method, angiogenic factors by immunoassay and JAK2V617F, and CALR mutations by DNA sequencing and allelic PCR. MVD was significantly increased in MPNs compared to controls (PMF > PV > ET). Correlation between MVD and plasma angiogenic factors was found in MPNs. MVD was significantly increased in patients with JAK2V617F mutation and correlated with JAK2 mutant allele burden (CD34-MVD: ρ = 0.491, p < 0.001; CD105-MVD: ρ = 0.276, p = 0.02) but not with CALR mutation. MVD correlated with leukocyte count, serum lactate dehydrogenase, hepatomegaly, and splenomegaly. BM fibrosis was significantly associated with CD34-MVD, CD105-MVD, interleukin-8, and JAK2 mutant allele burden. JAK2 homozygote status had positive predictive value (100%) for BM fibrosis. Patients with prefibrotic PMF had significantly higher MVD than patients with ET, and we could recommend MVD to be additional histopathological marker to distinguish these two entities. This study also highlights the strong correlation of MVD with plasma angiogenic factors, JAK2 mutant allele burden, and BM fibrosis in MPNs.

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Acknowledgements

This work was supported by a grant from Ministry of education, science, and technological development of the Republic of Serbia (175053) and Swiss National Science Foundation through Joint research project (SCOPES) IZ73Z0_152420/1. We are grateful to pathologists Maja Perunicic-Jovanovic, Tatjana Terzic, and Ljubomir Jakovic who have done histolopathological analysis and evaluation of bone marrow fibrosis.

Author contribution

Danijela Lekovic, Mirjana Gotic, Radek Skoda, and Vladan Cokic designed the study, performed the research, analyzed data and wrote manuscript. Bojana Cokic- Beleslin, Olivera Mitrovic-Ajtic, Dijana Sefer, Tijana Suboticki, Dragana Markovic, Marijana Buac, Milos Diklic, and Ronny Nienhold performed research. Natasa Milic performed statistical analyses.

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Authors and Affiliations

  1. Clinic for Hematology, Clinical Center of Serbia, Koste Todorovica 2, Belgrade, 11000, Serbia

    Danijela Lekovic, Mirjana Gotic & Dijana Sefer

  2. Medical Faculty, University of Belgrade, Belgrade, Serbia

    Danijela Lekovic, Mirjana Gotic & Natasa Milic

  3. Department of Biomedicine, Experimental Hematology, University Hospital Basel and University of Basel, Basel, Switzerland

    Radek Skoda & Ronny Nienhold

  4. Clinic for Endocrinology, Diabetes and Metabolic Diseases, Genetic laboratory, Clinical Center of Serbia, Belgrade, Serbia

    Bojana Beleslin-Cokic

  5. Institute for Medical Statistics, University of Belgrade, Belgrade, Serbia

    Natasa Milic

  6. Institute for Medical Research, University of Belgrade, Belgrade, Serbia

    Olivera Mitrovic-Ajtic, Tijana Suboticki, Marijana Buac, Dragana Markovic, Milos Diklic & Vladan P. Cokic

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  1. Danijela Lekovic
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Correspondence to Danijela Lekovic.

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Informed consent was obtained from all patients for being included in the study.

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Electronic supplementary material

Supplemental Table 1.

List of antibodies used for analyzing microvessel density (DOCX 11 kb)

Supplemental table 2.

The following primers were used for allelic PCR (DOCX 11 kb)

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Lekovic, D., Gotic, M., Skoda, R. et al. Bone marrow microvessel density and plasma angiogenic factors in myeloproliferative neoplasms: clinicopathological and molecular correlations. Ann Hematol 96, 393–404 (2017). https://doi.org/10.1007/s00277-016-2890-9

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  • DOI: https://doi.org/10.1007/s00277-016-2890-9

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