Modelization of Blood-Borne Hypercoagulability in Myeloma: A Tissue-Factor-Bearing Microparticle-Driven Process

Abstract

Introduction Hypercoagulability is a common blood alteration in newly diagnosed chemotherapy naïve patients with multiple myeloma. The identification of the procoagulant potential of cancer cells, which is principally related to tissue factor (TF) expression, attracts particular interest. The mechanisms by which myeloma plasma cells (MPCs) activate blood coagulation have been poorly investigated.

Aim To identify the principal actors related with MPCs that boost thrombin generation (TG).

Methods TF and annexin V expression by MPCs and MPC-derived microparticles (MPC-dMPs) was analyzed by flow cytometry. TF activity (TFa) and TF gene expression were also determined. TG in the presence of MPCs or MPC-dMPs was assessed with the calibrated automated thrombogram assay (CAT) in normal human PPP and in plasma depleted of factor VII or XII. TG was also assessed in plasma spiked with MPCs and MPC-dMPs.

Results MPC-dMPs expressed approximately twofold higher levels of TF as compared with MPCs. The TFa expressed by MPC-dMPs was significantly higher compared with that expressed by MPCs. MPCs and MPC-dMPs enhanced TG of human plasma. TG was significantly higher with MPC-dMPs compared with MPCs.

Conclusion MPCs indirectly induce blood-borne hypercoagulability through the release of MPC-dMPs rich in TF. Since MPCs, expressing low TFa, represent a weak procoagulant stimulus, the hypercoagulability at the microenvironment could be the resultant of MPC-dMPs rich in TF.

Keywords

multiple myeloma - thrombin generation - tissue factor - microparticles - hypercoagulability

Authors' Contributions

L.P. did the experiments, has made substantial contributions to study design and organization, acquisition, analysis, and interpretation of data, and has been involved in drafting the manuscript. K.A.H. had substantial contribution in analysis and interpretation of the data from flow cytometry measurements. S.T. did the PCR experiments and interpreted the corresponding results. E.M. did the cultures of the cancer cells and prepared the samples with microparticles. H.C. was involved in the cultures of the cancer cells and thrombin generation experiments. L.G. contributed to the design of the study and the acquisition of the MPC cultures. I.E. was involved in the critical revision of the manuscript.

A.L. critically revised the manuscript. P.V.D. has made substantial contribution to the design of the study, the interpretation of the data, and the writing and editing of the manuscript. M.A.D. critically revised the manuscript. M.M. contributed to critical revision of the manuscript. G.T.G. as the principal investigator has made substantial contributions to conception and design of the study, analysis, and interpretation of data, was involved in drafting the manuscript, gave the final approval of the version to be published, and agreed to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.

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