Cell-based evaluation of changes in coagulation activity induced by antineoplastic drugs for the treatment of acute myeloid leukemia

Tsunaka, Misae; Shinki, Haruka; Koyama, Takatoshi

doi:10.1371/journal.pone.0175765

Cited by 6 publications

(5 citation statements)

References 25 publications

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“…Early characterisation studies showed that U937 cells differed from typical lymphoblastoid cell lines [42]. Because of their pro-monocytic phenotype, many regard the U937 cell line to be representative of an AML subtype M5, under the French-American-British (FAB) classification system [43][44][45][46]. This pro-monocytic phenotype and cytochemistry, accounted for the wide use of U937 cells as a model to study AML, including ROS mediated monocyte to macrophage transition [32,47,48].…”

Section: Discussionmentioning

confidence: 99%

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The BCAT1 CXXC Motif Provides Protection against ROS in Acute Myeloid Leukaemia Cells

et al. 2022

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The cytosolic branched-chain aminotransferase (BCAT1) has received attention for its role in myeloid leukaemia development, where studies indicate metabolic adaptations due to BCAT1 up-regulation. BCAT1, like the mitochondria isoform (BCAT2), shares a conserved CXXC motif ~10 Å from the active site. This CXXC motif has been shown to act as a ‘redox-switch’ in the enzymatic regulation of the BCAT proteins, however the response to reactive oxygen species (ROS) differs between BCAT isoforms. Studies indicate that the BCAT1 CXXC motif is several orders of magnitude less sensitive to the effects of ROS compared with BCAT2. Moreover, estimation of the reduction mid-point potential of BCAT1, indicates that BCAT1 is more reductive in nature and may possess antioxidant properties. Therefore, the aim of this study was to further characterise the BCAT1 CXXC motif and evaluate its role in acute myeloid leukaemia. Our biochemical analyses show that purified wild-type (WT) BCAT1 protein could metabolise H2O2 in vitro, whereas CXXC motif mutant or WT BCAT2 could not, demonstrating for the first time a novel antioxidant role for the BCAT1 CXXC motif. Transformed U937 AML cells over-expressing WT BCAT1, showed lower levels of intracellular ROS compared with cells over-expressing the CXXC motif mutant (CXXS) or Vector Controls, indicating that the BCAT1 CXXC motif may buffer intracellular ROS, impacting on cell proliferation. U937 AML cells over-expressing WT BCAT1 displayed less cellular differentiation, as observed by a reduction of the myeloid markers; CD11b, CD14, CD68, and CD36. This finding suggests a role for the BCAT1 CXXC motif in cell development, which is an important pathological feature of myeloid leukaemia, a disease characterised by a block in myeloid differentiation. Furthermore, WT BCAT1 cells were more resistant to apoptosis compared with CXXS BCAT1 cells, an important observation given the role of ROS in apoptotic signalling and myeloid leukaemia development. Since CD36 has been shown to be Nrf2 regulated, we investigated the expression of the Nrf2 regulated gene, TrxRD1. Our data show that the expression of TrxRD1 was downregulated in transformed U937 AML cells overexpressing WT BCAT1, which taken with the reduction in CD36 implicates less Nrf2 activation. Therefore, this finding may implicate the BCAT1 CXXC motif in wider cellular redox-mediated processes. Altogether, this study provides the first evidence to suggest that the BCAT1 CXXC motif may contribute to the buffering of ROS levels inside AML cells, which may impact ROS-mediated processes in the development of myeloid leukaemia.

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Section: Discussionmentioning

confidence: 99%

“…However, our study is limited to a single FAB subtype model. Future work could extend our analysis and examine the antioxidant capacity of the BCAT1 CXXC motif in other FAB subtype models, for example HL60 (AML, M2) and NB4 (AML M3 aka APL) [45].…”

Section: Discussionmentioning

confidence: 99%

The BCAT1 CXXC Motif Provides Protection against ROS in Acute Myeloid Leukaemia Cells

et al. 2022

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“…A portion of target cells (2 × 10 6 ) was suspended in 50 µL phosphate-buffered saline (PBS) and added to 50 µL of healthy human plasma. After the incubation at 37 • C for 3 min, 50 µL of 25 mM calcium chloride was added, and the plasma recalcification time was measured using a semi-automatic coagulator (CA-50, Sysmex, Kobe, Japan) [15]. CA-50 detected changes in plasma optical density during blood coagulation.…”

Section: Procoagulant Activity (Pca) Assaymentioning

confidence: 99%

Interferon-γ Produced by EBV-Positive Neoplastic NK-Cells Induces Differentiation into Macrophages and Procoagulant Activity of Monocytes, Which Leads to HLH

Yoshimori

Nishio

Ohashi

et al. 2021

Cancers

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Epstein–Barr virus (EBV)-positive T- or NK-cell neoplasms show progressive systemic inflammation and abnormal blood coagulation causing hemophagocytic lymphohistiocytosis (HLH). It was reported that inflammatory cytokines were produced and secreted by EBV-positive neoplastic T- or NK-cells. These cytokines can induce the differentiation of monocytes into macrophages leading to HLH. To clarify which products of EBV-positive neoplastic T- or NK-cells have effects on monocytes, we performed a co-culture assay of monocytes with the supernatants of EBV-positive T- or NK-cell lines. The expression of differentiation markers, the phagocytosis ability, and the mRNA expression of the inflammatory cytokines of THP-1, a monocytic cell line, clearly increased after culturing with the supernatants from EBV-NK-cell lines. Co-culturing with the supernatants promoted the expression of CD80 and CD206 as well as M1 and M2 macrophage markers in human monocytes. Co-culturing with the supernatants of EBV-NK-cell lines significantly enhanced the procoagulant activity and the tissue factor expression of monocytes. Interferon (IFN)-γ was elevated extremely not only in the supernatant of EBV-NK-cell lines but also in the plasma of EBV-positive NK-cell neoplasms patients accompanying HLH. Finally, we confirmed that IFN-γ directly enhanced the differentiation into M1-like macrophages and the procoagulant activity of monocytes. Our findings suggest that IFN-γ may potentially serve as a therapeutic target to regulate HLH in EBV-positive NK-cell neoplasms.

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“…Было показано, что при инкубации клеточных культур клеток эндотелия (EAhy926), а также опухолевых клеточных ОМЛ-линий (HL60 (ОМЛ М2), NB4 (ОМЛ М3) и U9379 (ОМЛ М5)) с идарубицином в течение суток происходит укорочение времени свертывания пулированной плазмы здоровых добровольцев (в качестве активатора используется суспензия клеточной культуры). Также инкубация с идарубицином приводила к повышению экспрессии ТФ, тромбомодулина и фосфатидилсерина на поверхности клеток исследуемых клеточных линий [40]. Цитозин-арабинозид не оказывал влияния на исследуемые клеточные линии [40].…”

Section: острый миелобластный лейкозunclassified

“…Также инкубация с идарубицином приводила к повышению экспрессии ТФ, тромбомодулина и фосфатидилсерина на поверхности клеток исследуемых клеточных линий [40]. Цитозин-арабинозид не оказывал влияния на исследуемые клеточные линии [40]. Дополнительная терапия кортикостероидами также может оказывать прокоагулянтное влияние на систему гемостаза, однако эти данные подтверждаются только отдельными исследованиями и не подтверждены с помощью метаанализа.…”

Section: острый миелобластный лейкозunclassified

Modern aspects of the pathogenesis, diagnosis and therapy of hemostasis disorders in children with acute leukemias

Koltsova¹,

Баландина²,

Seregina³

et al. 2019

Ross. ž. det. gematol. onkol.

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Patients with oncohematological diseases, both children and adults, face high risks of thrombotic and hemorrhagic complications.About 40 % of pediatric patients with acute lymphoblastic leukemia develop bleedings, and the incidence of thrombosis in this disease ranges from 1 to 36 %. Most thromboses are associated with the use of central venous catheters and the use of L-asparaginase, which leads to a significant reduction in the synthesis of coagulation proteins.Massive hemorrhages account for two-thirds of all causes of early death in pediatric patients with acute myelogenous leukemia (AML). Absolute risks of death due to bleeding and leukostasis range from 1.8 % in the total population of children with AML to 14.3 % in a population with hyperleukocytosis more than 200 × 109 /l. The risk of thrombotic complications in children with AML varies between 3.4–11 %. In patients with AML, complex systemic coagulopathies may occur, such as disseminated intravascular coagulation (DIC), excessive fibrinolysis, or nonspecific proteolysis. This scale is not yet applicable due to the lack of research on its effectiveness in the pediatric population. The laboratory diagnostics of hemostasis is difficult due to the combined nature of thrombotic and hemorrhagic complications: bleeding, thrombosis and even DIC syndrome (combining both hyper- and hypocoagulation phases) can be expected in each specific patient with hemoblastosis. Because of the long-term nature of the treatment and the varying intensity of the various treatment units, the patient’s hemostasis during disease manifestation does not allow one to predict with any certainty the complications on induction or consolidation therapy. Involving all the components of the hemostasis system – vascular, platelet and plasma – into the pathological process makes prediction and diagnosis of thrombohemorrhagic complications impossible with the help of standard hemostatic tests and a general blood test, since these tests are designed to assess the concentrations of individual proteins and the functioning of individual components of the hemostatic system, and does not assess the balance between its procoagulant and anticoagulant components. Global hemostatic tests such as thromboelastography, thrombodynamics and thrombin generation test adequately reflect hypercoagulable conditions and can serve as a basis for the development of a new set of laboratory hemostasis tests.Conflict of interest. F.I. Ataullakhanov is co-founder of HemaCore LLC, which holds several patents and patent applications that are related to the diagnostic use of Thrombodynamics® (Ataullakhanov F.I., international patent applications: PCT/CH2007/000543 filing date 02.11.2007 and РСТ/RU2012/000570 filing date 16.07.2012). None of the other authors has any competing interests to declare.

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Cell-based evaluation of changes in coagulation activity induced by antineoplastic drugs for the treatment of acute myeloid leukemia

Cited by 6 publications

References 25 publications

The BCAT1 CXXC Motif Provides Protection against ROS in Acute Myeloid Leukaemia Cells

The BCAT1 CXXC Motif Provides Protection against ROS in Acute Myeloid Leukaemia Cells

Interferon-γ Produced by EBV-Positive Neoplastic NK-Cells Induces Differentiation into Macrophages and Procoagulant Activity of Monocytes, Which Leads to HLH

Modern aspects of the pathogenesis, diagnosis and therapy of hemostasis disorders in children with acute leukemias

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