Identification of signalling cascades involved in red blood cell shrinkage and vesiculation

Elena B Kostova, Boukje M Beuger, Thomas R L Klei, Pasi Halonen, Cor Lieftink, Roderick Beijersbergen, Timo K van den Berg, Robin van Bruggen

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Even though red blood cell (RBC) vesiculation is a well-documented phenomenon, notably in the context of RBC aging and blood transfusion, the exact signalling pathways and kinases involved in this process remain largely unknown. We have established a screening method for RBC vesicle shedding using the Ca(2+) ionophore ionomycin which is a rapid and efficient method to promote vesiculation. In order to identify novel pathways stimulating vesiculation in RBC, we screened two libraries: the Library of Pharmacologically Active Compounds (LOPAC) and the Selleckchem Kinase Inhibitor Library for their effects on RBC from healthy donors. We investigated compounds triggering vesiculation and compounds inhibiting vesiculation induced by ionomycin. We identified 12 LOPAC compounds, nine kinase inhibitors and one kinase activator which induced RBC shrinkage and vesiculation. Thus, we discovered several novel pathways involved in vesiculation including G protein-coupled receptor (GPCR) signalling, the phosphoinositide 3-kinase (PI3K)-Akt (protein kinase B) pathway, the Jak-STAT (Janus kinase-signal transducer and activator of transcription) pathway and the Raf-MEK (mitogen-activated protein kinase kinase)-ERK (extracellular signal-regulated kinase) pathway. Moreover, we demonstrated a link between casein kinase 2 (CK2) and RBC shrinkage via regulation of the Gardos channel activity. In addition, our data showed that inhibition of several kinases with unknown functions in mature RBC, including Alk (anaplastic lymphoma kinase) kinase and vascular endothelial growth factor receptor 2 (VEGFR-2), induced RBC shrinkage and vesiculation.

Original languageEnglish
JournalFrontiers in Bioscience
Volume35
Issue number2
DOIs
Publication statusPublished - 16 Apr 2015

Cite this

Kostova, E. B., Beuger, B. M., Klei, T. R. L., Halonen, P., Lieftink, C., Beijersbergen, R., ... van Bruggen, R. (2015). Identification of signalling cascades involved in red blood cell shrinkage and vesiculation. Frontiers in Bioscience, 35(2). https://doi.org/10.1042/BSR20150019
Kostova, Elena B ; Beuger, Boukje M ; Klei, Thomas R L ; Halonen, Pasi ; Lieftink, Cor ; Beijersbergen, Roderick ; van den Berg, Timo K ; van Bruggen, Robin. / Identification of signalling cascades involved in red blood cell shrinkage and vesiculation. In: Frontiers in Bioscience. 2015 ; Vol. 35, No. 2.
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abstract = "Even though red blood cell (RBC) vesiculation is a well-documented phenomenon, notably in the context of RBC aging and blood transfusion, the exact signalling pathways and kinases involved in this process remain largely unknown. We have established a screening method for RBC vesicle shedding using the Ca(2+) ionophore ionomycin which is a rapid and efficient method to promote vesiculation. In order to identify novel pathways stimulating vesiculation in RBC, we screened two libraries: the Library of Pharmacologically Active Compounds (LOPAC) and the Selleckchem Kinase Inhibitor Library for their effects on RBC from healthy donors. We investigated compounds triggering vesiculation and compounds inhibiting vesiculation induced by ionomycin. We identified 12 LOPAC compounds, nine kinase inhibitors and one kinase activator which induced RBC shrinkage and vesiculation. Thus, we discovered several novel pathways involved in vesiculation including G protein-coupled receptor (GPCR) signalling, the phosphoinositide 3-kinase (PI3K)-Akt (protein kinase B) pathway, the Jak-STAT (Janus kinase-signal transducer and activator of transcription) pathway and the Raf-MEK (mitogen-activated protein kinase kinase)-ERK (extracellular signal-regulated kinase) pathway. Moreover, we demonstrated a link between casein kinase 2 (CK2) and RBC shrinkage via regulation of the Gardos channel activity. In addition, our data showed that inhibition of several kinases with unknown functions in mature RBC, including Alk (anaplastic lymphoma kinase) kinase and vascular endothelial growth factor receptor 2 (VEGFR-2), induced RBC shrinkage and vesiculation.",
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Kostova, EB, Beuger, BM, Klei, TRL, Halonen, P, Lieftink, C, Beijersbergen, R, van den Berg, TK & van Bruggen, R 2015, 'Identification of signalling cascades involved in red blood cell shrinkage and vesiculation' Frontiers in Bioscience, vol. 35, no. 2. https://doi.org/10.1042/BSR20150019

Identification of signalling cascades involved in red blood cell shrinkage and vesiculation. / Kostova, Elena B; Beuger, Boukje M; Klei, Thomas R L; Halonen, Pasi; Lieftink, Cor; Beijersbergen, Roderick; van den Berg, Timo K; van Bruggen, Robin.

In: Frontiers in Bioscience, Vol. 35, No. 2, 16.04.2015.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Identification of signalling cascades involved in red blood cell shrinkage and vesiculation

AU - Kostova, Elena B

AU - Beuger, Boukje M

AU - Klei, Thomas R L

AU - Halonen, Pasi

AU - Lieftink, Cor

AU - Beijersbergen, Roderick

AU - van den Berg, Timo K

AU - van Bruggen, Robin

PY - 2015/4/16

Y1 - 2015/4/16

N2 - Even though red blood cell (RBC) vesiculation is a well-documented phenomenon, notably in the context of RBC aging and blood transfusion, the exact signalling pathways and kinases involved in this process remain largely unknown. We have established a screening method for RBC vesicle shedding using the Ca(2+) ionophore ionomycin which is a rapid and efficient method to promote vesiculation. In order to identify novel pathways stimulating vesiculation in RBC, we screened two libraries: the Library of Pharmacologically Active Compounds (LOPAC) and the Selleckchem Kinase Inhibitor Library for their effects on RBC from healthy donors. We investigated compounds triggering vesiculation and compounds inhibiting vesiculation induced by ionomycin. We identified 12 LOPAC compounds, nine kinase inhibitors and one kinase activator which induced RBC shrinkage and vesiculation. Thus, we discovered several novel pathways involved in vesiculation including G protein-coupled receptor (GPCR) signalling, the phosphoinositide 3-kinase (PI3K)-Akt (protein kinase B) pathway, the Jak-STAT (Janus kinase-signal transducer and activator of transcription) pathway and the Raf-MEK (mitogen-activated protein kinase kinase)-ERK (extracellular signal-regulated kinase) pathway. Moreover, we demonstrated a link between casein kinase 2 (CK2) and RBC shrinkage via regulation of the Gardos channel activity. In addition, our data showed that inhibition of several kinases with unknown functions in mature RBC, including Alk (anaplastic lymphoma kinase) kinase and vascular endothelial growth factor receptor 2 (VEGFR-2), induced RBC shrinkage and vesiculation.

AB - Even though red blood cell (RBC) vesiculation is a well-documented phenomenon, notably in the context of RBC aging and blood transfusion, the exact signalling pathways and kinases involved in this process remain largely unknown. We have established a screening method for RBC vesicle shedding using the Ca(2+) ionophore ionomycin which is a rapid and efficient method to promote vesiculation. In order to identify novel pathways stimulating vesiculation in RBC, we screened two libraries: the Library of Pharmacologically Active Compounds (LOPAC) and the Selleckchem Kinase Inhibitor Library for their effects on RBC from healthy donors. We investigated compounds triggering vesiculation and compounds inhibiting vesiculation induced by ionomycin. We identified 12 LOPAC compounds, nine kinase inhibitors and one kinase activator which induced RBC shrinkage and vesiculation. Thus, we discovered several novel pathways involved in vesiculation including G protein-coupled receptor (GPCR) signalling, the phosphoinositide 3-kinase (PI3K)-Akt (protein kinase B) pathway, the Jak-STAT (Janus kinase-signal transducer and activator of transcription) pathway and the Raf-MEK (mitogen-activated protein kinase kinase)-ERK (extracellular signal-regulated kinase) pathway. Moreover, we demonstrated a link between casein kinase 2 (CK2) and RBC shrinkage via regulation of the Gardos channel activity. In addition, our data showed that inhibition of several kinases with unknown functions in mature RBC, including Alk (anaplastic lymphoma kinase) kinase and vascular endothelial growth factor receptor 2 (VEGFR-2), induced RBC shrinkage and vesiculation.

KW - Calcium Ionophores/pharmacology

KW - Casein Kinase II/metabolism

KW - Cell-Derived Microparticles/metabolism

KW - Cellular Senescence/drug effects

KW - Erythrocytes/cytology

KW - Humans

KW - Ionomycin/pharmacology

KW - raf Kinases/metabolism

U2 - 10.1042/BSR20150019

DO - 10.1042/BSR20150019

M3 - Article

VL - 35

JO - Frontiers in Bioscience

JF - Frontiers in Bioscience

SN - 1093-9946

IS - 2

ER -

Kostova EB, Beuger BM, Klei TRL, Halonen P, Lieftink C, Beijersbergen R et al. Identification of signalling cascades involved in red blood cell shrinkage and vesiculation. Frontiers in Bioscience. 2015 Apr 16;35(2). https://doi.org/10.1042/BSR20150019