A systems-biology analysis of isogenic megakaryocytic and granulocytic cultures identifies new molecular components of megakaryocytic apoptosis

Chen, Chi; Fuhrken, Peter G.; Huang, Li Chi; Apostolidis, Pani A.; Wang, Min; Paredes, Carlos J.; Miller, William M.; Papoutsakis, Eleftherios T.

doi:10.1186/1471-2164-8-384

Cited by 18 publications

(26 citation statements)

References 75 publications

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“…We have recently shown that GADD45a is up-regulated in human megakaryocytic cells undergoing terminal differentiation (41). However, we have not yet validated a functional role of GADD45a on megakaryocytic differentiation.…”

Section: P53 In Mk Differentiationmentioning

confidence: 92%

Tumor Suppressor Protein p53 Regulates Megakaryocytic Polyploidization and Apoptosis

Fuhrken

Apostolidis

Lindsey

et al. 2008

Journal of Biological Chemistry

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The molecular mechanisms underlying differentiation of hematopoietic stem cells into megakaryocytes are poorly understood. Tumor suppressor protein p53 can act as a transcription factor affecting both cell cycle control and apoptosis, and we have previously shown that p53 is activated during terminal megakaryocytic (Mk) differentiation of the CHRF-288-11 (CHRF) cell line. Here, we use RNA interference to reduce p53 expression in CHRF cells and show that reduced p53 activity leads to a greater fraction of polyploid cells, higher mean and maximum ploidy, accelerated DNA synthesis, and delayed apoptosis and cell death upon phorbol 12-myristate 13-acetate-induced Mk differentiation. In contrast, reduced p53 expression did not affect the ploidy or DNA synthesis of CHRF cells in the absence of phorbol 12-myristate 13-acetate stimulation. Furthermore, primary Mk cells from cultures initiated with p53-null mouse bone marrow mononuclear cells displayed higher ploidy compared with wild-type controls. Quantitative reverse transcription-PCR analysis of p53-knockdown CHRF cells, compared with the "scrambled" control CHRF cells, revealed that six known transcriptional targets of p53 (BBC3, BAX, TP53I3, TP53INP1, MDM2, and P21) were down-regulated, whereas BCL2 expression, which is known to be negatively affected by p53, was up-regulated. These studies show that the functional role of the intrinsic activation of p53 during Mk differentiation is to control polyploidization and the transition to endomitosis by impeding cell cycling and promoting apoptosis.

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Section: P53 In Mk Differentiationmentioning

confidence: 92%

Tumor Suppressor Protein p53 Regulates Megakaryocytic Polyploidization and Apoptosis

Fuhrken

Apostolidis

Lindsey

et al. 2008

Journal of Biological Chemistry

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“…Thus, immunofluorescence coupled to mean fluorescent intensity quantification was chosen for quantitating the levels of several proteins, a well established method for quantitating protein expression on a single-cell basis (34 -36). Immunofluorescence staining was performed as described by Kodiha et al (37) (38 -42), as well as from our own laboratory (43)(44)(45). Quantification was performed within the cell perimeter only; to delineate the cell borders, 5 M Syto13 (Life Technologies number S7575) or Syto40 (Life Technologies number S11351) was used.…”

Section: Megakaryocyte Cultures and Exposure Of Mk Cells To Shearmentioning

confidence: 99%

Megakaryocytic Maturation in Response to Shear Flow Is Mediated by the Activator Protein 1 (AP-1) Transcription Factor via Mitogen-activated Protein Kinase (MAPK) Mechanotransduction

Luff

Papoutsakis

2016

Journal of Biological Chemistry

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Megakaryocytes (MKs) are exposed to shear flow as they migrate from the bone marrow hematopoietic compartment into circulation to release pro/preplatelets into circulating blood. Shear forces promote DNA synthesis, polyploidization, and maturation in MKs, and platelet biogenesis. To investigate mechanisms underlying these MK responses to shear, we carried out transcriptional analysis on immature and mature stem cell-derived MKs exposed to physiological shear. In immature (day (d)9) MKs, shear exposure up-regulated genes related to growth and MK maturation, whereas in mature (d12) MKs, it up-regulated genes involved in apoptosis and intracellular transport.

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“…We mined the RNA sequencing data for genes already defined by others as megakaryocyte-specific and genes highly expressed during megakaryocyte differentiation. 23,24 Of the 56 genes curated by the Papoutsakis group as being megakaryocyte related by literature review, 24 were deregulated by loss of both MKL1 and …”

Section: Rna Sequencing Reveals Srf-independent Functions For Mrtfs Imentioning

confidence: 99%

MKL1 and MKL2 play redundant and crucial roles in megakaryocyte maturation and platelet formation

Smith

Thon

Devine

et al. 2012

Blood

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A systems-biology analysis of isogenic megakaryocytic and granulocytic cultures identifies new molecular components of megakaryocytic apoptosis

Cited by 18 publications

References 75 publications

Tumor Suppressor Protein p53 Regulates Megakaryocytic Polyploidization and Apoptosis

Tumor Suppressor Protein p53 Regulates Megakaryocytic Polyploidization and Apoptosis

Megakaryocytic Maturation in Response to Shear Flow Is Mediated by the Activator Protein 1 (AP-1) Transcription Factor via Mitogen-activated Protein Kinase (MAPK) Mechanotransduction

MKL1 and MKL2 play redundant and crucial roles in megakaryocyte maturation and platelet formation

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