Interrelation between polyploidization and megakaryocyte differentiation: a gene profiling approach

Raslová, Hana; Kauffmann, Audrey; Sekkaı̈, Dalila; Ripoche, Hugues; Larbret, Frédéric; Thomas, Robert; Roux, Diana Tronik‐Le; Kroemer, Guido; Debili, Najet; Dessen, Philippe; Lazar, Vladimir; Vainchenker, William

doi:10.1182/blood-2006-07-037838

Cited by 99 publications

(98 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Validation of the proteomics with antibodies against key proteins, including NMM-IIA and vinculin (Fig. S7D), proved consistent with mRNA up-regulation of NMM-IIA and vinculin in MKs (30).…”

Section: Results and Analysissupporting

confidence: 55%

Myosin-II inhibition and soft 2D matrix maximize multinucleation and cellular projections typical of platelet-producing megakaryocytes

Shin

Swift

Spinler

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Cell division, membrane rigidity, and strong adhesion to a rigid matrix are all promoted by myosin-II, and so multinucleated cells with distended membranes-typical of megakaryocytes (MKs)-seem predictable for low myosin activity in cells on soft matrices. Paradoxically, myosin mutations lead to defects in MKs and platelets. Here, reversible inhibition of myosin-II is sustained over several cell cycles to produce 3-to 10-fold increases in polyploid MK and a number of other cell types. Even brief inhibition generates highly distensible, proplatelet-like projections that fragment readily under shear, as seen in platelet generation from MKs in vivo. The effects are maximized with collagenous matrices that are soft and 2D, like the perivascular niches in marrow rather than 3D or rigid, like bone. Although multinucleation of other primary hematopoietic lineages helps to generalize a failure-to-fission mechanism, lineage-specific signaling with increased polyploidy proves possible and novel with phospho-regulation of myosin-II heavy chain. Labelfree mass spectrometry quantitation of the MK proteome uses a unique proportional peak fingerprint (ProPF) analysis to also show upregulation of the cytoskeletal and adhesion machinery critical to platelet function. Myosin-inhibited MKs generate more platelets in vitro and also in vivo from the marrows of xenografted mice, while agonist stimulation activates platelet spreading and integrin αIIbβ3. Myosin-II thus seems a central, matrix-regulated node for MK-poiesis and platelet generation. matrix elasticity | blebbistatin | proteomics | thrombocytopenia

show abstract

Section: Results and Analysissupporting

confidence: 55%

Myosin-II inhibition and soft 2D matrix maximize multinucleation and cellular projections typical of platelet-producing megakaryocytes

Shin

Swift

Spinler

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…We previously found by gene-profiling assays that p19 INK4D expression was increasing linearly during ploidization. 12 Such a result was not found for other CDKIs belonging to either the Cip/Kip or INK4 families, whether the analyses were performed by microarrays 12 or by quantitative real-time RT-PCR ( Figure 1C). In addition, our data ( Figure 1D) show that the p19 INK4D protein also increased with ploidization, except at the 4N stage.…”

Section: Discussionmentioning

confidence: 84%

“…11 By performing a transcriptional profile of genes regulated during MK ploidization, we have shown that, among the CDKI, only p19 INK4D mRNA was linearly regulated through the 2N to 16N stages. 12 Submitted September 17, 2007; accepted February 6, 2008. Prepublished online as Blood First Edition paper, February 14, 2008 DOI 10.1182 DOI 10.…”

mentioning

confidence: 99%

P19INK4D links endomitotic arrest and megakaryocyte maturation and is regulated by AML-1

Gilles

Guièze

Bluteau

et al. 2008

Blood

Self Cite

View full text Add to dashboard Cite

The molecular mechanisms that regulate megakaryocyte (MK) ploidization are poorly understood. Using MK differentiation from primary human CD34 ؉ cells, we observed that p19 INK4D expression was increased both at the mRNA and protein levels during ploidization. p19 INK4D knockdown led to a moderate increase (31.7% ؎ 5%) in the mean ploidy of MKs suggesting a role of p19 INK4D in the endomitotic arrest. This increase in ploidy was associated with a decrease in the more mature MK population (CD41 high CD42 high ) at day 9 of culture, which was related to a delay in differentiation. Inversely, p19 INK4D Until now, the best-studied model of polyploidization concerns megakaryocytes (MKs) in which after several replicative rounds, the process of classic mitosis is replaced by an endomitotic process. MK endomitosis is very similar to mitosis, but MKs skip anaphase B, telophase, and cytokinesis giving rise to a polyploid cell with a single polylobulated nucleus. 1,2 Although polyploidization is a part of the MK differentiation program, terminal differentiation that is associated with an increased platelet protein synthesis and the development of specific organelles requires an arrest of the cell cycle. Both the mitotic and endomitotic processes require an amplification of the DNA content. However, while mitosis leads to an increase in cell number, polyploidization results in an increase in the cell size and protein mass. In the case of MK, the platelet precursor, polyploidization increases platelet production.DNA replication is a regulated mechanism that depends on a well-organized network of the cyclin-dependent kinases (CDKs) and cyclin-dependent inhibitors (CDKIs) of the cell cycle. Progression through G1 to S phase of cell cycle in mammalian cells requires the activity of G1-cyclins (D and E) associated with their catalytic subunits (CDK4 and CDK6). The mitogen-stimulated cyclin D and CDK4/6 complexes phosphorylate the retinoblastoma protein (Rb) allowing its dissociation from E2Fs transcription factors. Activated E2Fs then regulate the expression of genes necessary for DNA synthesis during the S phase of cell cycle. Cells exit cell cycle and accumulate in a quiescent G 0 /G 1 state either in the absence of mitogenic stimuli when the cyclin D-dependent activity is lost or after CDK4/6 inhibition. CDKs are regulated by inhibitory phosphorylations mediated by the WEE1 and MYT1 kinases 3 or by induction of specific inhibitors from the INK4 and CIP/KIP families. Till now, 4 members of the INK4 family (p19 INK4D , p18 INK4C , p16 INK4A , and p15 INK4B ) are described to specifically block the activity of CDK4/6 either by forming inactive ternary INK4-CDK4/6-cyclin D or binary INK4-CDK4/6 complexes. 4 The members of the CIP/KIP family (p21 CIP1/WAF1 , p27 KIP1 , and p57 KIP2 ) inhibit cyclin A-or cyclin E-associated CDK2 activity, but stabilize cyclin D-associated CDK4/6 activity. 5,6 In MKs, a high level of 2 CIP/KIP family members, p21 CIP1 and p27 KIP1 ,7,8 and 1 member of the INK4 family, p16 INK4A , 7 was detected...

show abstract

“…Large MEGs are a result of endomitosis which allows an increase in cell mass without the need to devote energy to execute all aspects of mitosis 21. Furthermore, polyploidy differentially regulates genes needed for MEG maturation and platelet production 22. Thus, endomitosis results in higher differentiated cells that are capable of producing more platelets than a cell of lower ploidy 23.…”

Section: Discussionmentioning

confidence: 99%

Iron deficiency alters megakaryopoiesis and platelet phenotype independent of thrombopoietin

et al. 2014

View full text Add to dashboard Cite

Iron deficiency is a common cause of reactive thrombocytosis, however, the exact pathways have not been revealed. Here we aimed to study the mechanisms behind iron deficiency‐induced thrombocytosis. Within few weeks, iron‐depleted diet caused iron deficiency in young Sprague–Dawley rats, as reflected by a drop in hemoglobin, mean corpuscular volume, hepatic iron content and hepcidin mRNA in the liver. Thrombocytosis established in parallel. Moreover, platelets produced in iron deficient animals displayed a higher mean platelet volume and increased aggregation. Bone marrow studies revealed subtle alterations that are suggestive of expansion of megakaryocyte progenitors, an increase in megakaryocyte ploidy and accelerated megakaryocyte differentiation. Iron deficiency did not alter the production of hematopoietic growth factors such as thrombopoietin, interleukin 6 or interleukin 11. Megakaryocytic cell lines grown in iron‐depleted conditions exhibited reduced proliferation but increased ploidy and cell size. Our data suggest that iron deficiency increases megakaryopoietic differentiation and alters platelet phenotype without changes in megakaryocyte growth factors, specifically TPO. Iron deficiency‐induced thrombocytosis may have evolved to maintain or increase the coagulation capacity in conditions with chronic bleeding. Am. J. Hematol. 89:524–529, 2014. © 2014 Wiley Periodicals, Inc.

show abstract

Interrelation between polyploidization and megakaryocyte differentiation: a gene profiling approach

Cited by 99 publications

References 42 publications

Myosin-II inhibition and soft 2D matrix maximize multinucleation and cellular projections typical of platelet-producing megakaryocytes

Myosin-II inhibition and soft 2D matrix maximize multinucleation and cellular projections typical of platelet-producing megakaryocytes

P19INK4D links endomitotic arrest and megakaryocyte maturation and is regulated by AML-1

Iron deficiency alters megakaryopoiesis and platelet phenotype independent of thrombopoietin

Contact Info

Product

Resources

About