Pro- and Antifibrotic Factors in the Serum of Patients with Chronic Myeloproliferative Disorders

AA Silyutina, NM Matyukhina, EG Lisina, VI Khvan, SN Leleko, NT Siordiya, OV Sirotkina, PA Butylin

VA Almazov National Medical Research Center, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341

For correspondence: Pavel Andreevich Butylin, PhD, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341; e-mail: butylinp@gmail.com

For citation: Silyutina AA, Matyukhina NM, Lisina EG, et al. Pro- and Antifibrotic Factors in the Serum of Patients with Chronic Myeloproliferative Disorders. Clinical oncohematology. 2017;10(4):479–84 (In Russ).

DOI: 10.21320/2500-2139-2017-10-4-479-484


ABSTRACT

Background. The study of pro- and antifibrotic factors in the serum of patients with Ph-negative chronic myeloproliferative disorders (CMPDs) will allow to understand better the mechanisms of myelofibrosis development, as well as to identify new diagnostic markers.

Aim. To assess the correlation between the levels of classic (TGF-β, bFGF, MMP-2, -9, -13 and VEGF) and new proinflammatory serum factors (galectin-3), involved into development of myelofibrosis in different Ph-negative forms of CMPDs and genetic abnormalities.

Materials & Methods. The research included 55 CMPD patients (13 with polycythemia vera, 17 with essential thrombocythemia, 25 with primary myelofibrosis) and 8 healthy controls. Whole blood genomic DNA extraction was used to evaluate mutations JAK2V617F, CALR (deletions and insertions), MPLW515L, and MPLW515K. Antibody-immobilized ELISA was used to evaluate the levels of galectin-3, TGF-β, bFGF, VEGF, MMP-2, MMP-9 and MMP-13.

Results. The analysis showed the differences in serum MMP-9, VEGF, TGF-β and galectin-3 levels in patients with different CMPDs. A tendency towards the decrease of serum MMP-9 levels in patients with CALR mutations was shown.

Conclusion. The shown differences between patients with different CMPDs may serve as a basis for improving diagnostic protocols in challenging differential diagnosis of CMPDs.

Keywords: Ph-negative chronic myeloproliferative disorders, pro- and antifibrotic factors, JAK2V617F, CALR, MPLW515L, MPLW515K, MMP-2, MMP-9, MMP-13, galectin-3.

Received: April 26, 2017

Accepted: July 5, 2017

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Myelofibrosis Models: Literature Review and Own Data

AA Silyutina, II Gin, NM Matyukhina, EN Balayan, PA Butylin

VA Almazov Federal North-West Medical Research Center, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341

For correspondence: Pavel Andreevich Butylin, PhD, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341; e-mail: butylinp@gmail.com

For citation: Silyutina AA, Gin II, Matyukhina NM, et al. Myelofibrosis Models: Literature Review and Own Data. Clinical oncohematology. 2017;10(1):75–84 (In Russ).

DOI: 10.21320/2500-2139-2017-10-1-75-84


ABSTRACT

Background & Aims. Chronic myeloproliferative disorders typically develop during a long latent period, and it complicates the study of the mechanism of its pathogenesis. Observations from the clinical practice should be confirmed by experiments. The mechanisms of oncological transformation related to mutations associated with chronic myeloproliferative diseases were confirmed in transgene animal models. Biological models permitted to determine a complex nature of myelofibrosis. However, studies of the cellular mechanisms of myelofibrosis require new models. This paper presents a review of published models of myeloproliferative disorders, mainly, primary myelofibrosis, and results of studies of a new cell line with expression of JAK2 V617F. The aim of this study is to create a new cell line with expression of transforming JAK2 V617F mutation in acute monocytic leukemia THP-1 cells.

Methods. Transgenic cell lines were created on the basis of monocytic leukemia THP-1 cell line that can differentiate into macrophages. Direct mutagenesis was used to cause V617F mutation. Two cell lines were created: one with JAK2 expression with V617F mutation, the other with wild type JAK2.

Results. Both transgenic lines were characterized by increased JAK2 expression as compared to non-modified cells. In routine cultivation, transgenic THP-1 cells retained the morphology of monocytes. After treatment with phorbol ester, THP-1 differentiated into macrophages and become adherent to culture plastic. Adherent cells demonstrated the variety of shapes: some of them were spherical, the other ones had pseudopodia. No significant differences in viability of cells were observed. However, macrophages expressing mutant JAK2 and overexpressing the wild type JAK2 demonstrated a tendency to decreased amount unlivable cells during cultivation.

Conclusion. The obtained cell model can be used for estimating the influence of JAK2 V617F mutation on pro- and antifibrotic potential of macrophages that can help to investigate the pathogenetic role of macrophages in myelofibrosis development. In addition, this model can help to develop novel methods of therapy and diagnostics of primary and secondary myelofibrosis.

Keywords: Ph-negative chronic myeloproliferative disorders, primary myelofibrosis, JAK2 V617F, transgenic animals.

Received: September 15, 2016

Accepted: December 13, 2016

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