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Updated: Jan 31, 2026
Preparation of Washed Human Platelets for Quantitative Metabolic Flux Studies
Published on: January 10, 2025
Platelet expression of PKCepsilon oncoprotein in myelofibrosis is associated with disease severity and thrombotic
Elena Masselli1, Cecilia Carubbi1, Giulia Pozzi1
1Department of Medicine and Surgery, University of Parma, Ospedale Maggiore, Parma 43126, Italy.
Background:
Myelofibrosis (MF) is the most aggressive Philadelphia-negative chronic myeloproliferative neoplasm (MPN) with high morbidity and mortality due to thrombo-hemorrhagic complications and leukemic transformation. MF is characterized by profound alterations of megakaryocytopoiesis, with consequent abnormalities in platelet number and function. We recently showed that the overexpression of the oncoprotein PKCepsilon plays a key role in the aberrant differentiation of MF megakaryocyte clone and that its levels correlate with disease burden. Moreover, our group previously demonstrated that PKCepsilon is over-expressed in platelets from patients with acute myocardial infarction (MI) and accounts for their increased reactivity. On these bases, we investigated here the activation state and PKCepsilon expression of MF platelets, testing potential correlations with thrombotic risk and disease aggressiveness.
Methods:
Platelets were isolated from peripheral blood samples of MF patients and healthy donors (HDs). Patients were stratified according to the IPSS/DIPSS risk category and history of cardiovascular events. Platelet activation was assessed by flow cytometry. PKCepsilon mRNA and protein levels were determined by real time-PCR and western blot.
Results:
MF platelets circulate in an activated status and display significantly higher levels of PKCepsilon compared to HDs. In MF patients, PKCepsilon platelet levels were associated with high-risk disease as well as with a positive history of major cardiovascular events.
Conclusions:
PKCepsilon is configuring as the common denominator of neoplastic transformation and thrombus formation in MF. Overall, our data pinpoint PKCepsilon as a potential novel biomarker of disease aggressiveness and thrombotic risk in this hematologic neoplasm.
Insights
Platelets in myelofibrosis (MF) are activated and show high PKCepsilon levels, correlating with disease risk and cardiovascular events. This suggests PKCepsilon as a biomarker for MF aggressiveness and clotting risk.
Area of Science:
- Hematology
- Oncology
- Molecular Biology
Background:
- Myelofibrosis (MF) is an aggressive Philadelphia-negative chronic myeloproliferative neoplasm (MPN) associated with significant morbidity and mortality.
- MF involves aberrant megakaryocytopoiesis, leading to platelet count and function abnormalities.
- Overexpressed PKCepsilon is implicated in MF megakaryocyte differentiation and platelet hyper-reactivity in myocardial infarction.
Purpose of the Study:
- To investigate the activation status and PKCepsilon expression in platelets from MF patients.
- To explore the correlation between PKCepsilon levels, thrombotic risk, and disease aggressiveness in MF.
Main Methods:
- Peripheral blood platelets were isolated from MF patients and healthy donors (HDs).
- Patient risk stratification used IPSS/DIPSS categories and cardiovascular event history.
- Platelet activation was measured by flow cytometry; PKCepsilon levels by real-time PCR and Western blot.
Main Results:
- MF platelets exhibited an activated state and significantly elevated PKCepsilon levels compared to HDs.
- Higher PKCepsilon levels in MF patients correlated with high-risk disease.
- Elevated PKCepsilon was associated with a history of major cardiovascular events in MF patients.
Conclusions:
- PKCepsilon appears to be a shared factor in MF neoplastic transformation and thrombus formation.
- PKCepsilon is identified as a potential novel biomarker for assessing disease aggressiveness and thrombotic risk in myelofibrosis.
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