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MicroRNA expression profile in granulocytes from primary myelofibrosis patients
Paola Guglielmelli1, Lorenzo Tozzi, Alessandro Pancrazzi
1Department of Hematology, University of Florence, Florence, Italy.
Objective:
Expression profiling of microRNA (miRNA) was performed in granulocytes isolated from patients with primary myelofibrosis (PMF), with the aim of identifying abnormally expressed miRNAs in comparison with normal subjects or patients with polycythemia vera (PV) or essential thrombocythemia (ET).
Patients And Methods:
Using stem loop-primed reverse transcription and TaqMan quantitative real-time polymerase chain reaction, the expression of 156 mature miRNAs was evaluated using pooled granulocytes from PMF patients, either wild-type or JAK2(617V>F) mutant with >51% allele burden, and control subjects. Differentially expressed miRNAs were then validated on additional control and PMF samples, and also on PV or ET granulocytes.
Results:
There was a global downregulation of miRNA expression in PMF granulocytes; 60 miRNAs, of 128 called present, displayed differential expression compared to normal samples. Twelve miRNAs, which had been selected based on statistically different expression level, were finally validated. In PMF granulocytes, levels of miR-31, -150, and -95 were significantly lower, while those of miR-190 significantly greater, than control and PV or ET samples; on the other hand, miR-34a, -342, -326, -105, -149, and -147 were similarly reduced in patients with PMF, PV, or ET compared to controls. Increased expression of miR-182 and -183 correlated with JAK2(617V>F) allele burden. Three in silico-predicted putative target genes (DTR, HMGA2, and MYB), showed deregulated expression in PMF granulocytes that correlated with expression level of regulatory miRNA.
Conclusions:
A defined miRNA profile distinguishes PMF granulocytes from those of normal subjects and, partially, also from PV or ET patients.
Insights
MicroRNA (miRNA) expression is globally downregulated in primary myelofibrosis (PMF) granulocytes. A distinct miRNA profile differentiates PMF from normal and other myeloproliferative neoplasms.
Area of Science:
- Hematology
- Molecular Biology
- Genetics
Background:
- Primary myelofibrosis (PMF) is a myeloproliferative neoplasm characterized by bone marrow fibrosis.
- MicroRNAs (miRNAs) are small non-coding RNAs that play crucial roles in gene regulation and have been implicated in various cancers, including hematological malignancies.
Purpose of the Study:
- To investigate the differential expression of miRNAs in granulocytes from PMF patients compared to healthy controls and patients with polycythemia vera (PV) or essential thrombocythemia (ET).
- To identify specific miRNA signatures associated with PMF and potential correlations with JAK2 mutation status.
Main Methods:
- Expression profiling of 156 mature miRNAs using stem loop-primed reverse transcription and TaqMan quantitative real-time PCR in granulocytes.
- Validation of differentially expressed miRNAs in independent cohorts of PMF, PV, ET, and control samples.
- Correlation analysis of miRNA expression with JAK2(617V>F) allele burden and target gene expression.
Main Results:
- A global downregulation of miRNA expression was observed in PMF granulocytes.
- Sixty out of 128 detected miRNAs showed differential expression in PMF compared to normal samples.
- Specific miRNAs (e.g., miR-31, -150, -95, -190) exhibited distinct expression patterns in PMF granulocytes compared to controls and other myeloproliferative neoplasms. Increased expression of miR-182 and -183 correlated with JAK2(617V>F) allele burden.
Conclusions:
- A unique miRNA expression profile can distinguish PMF granulocytes from normal subjects and partially from PV or ET patients.
- These findings suggest that aberrant miRNA expression is a feature of PMF and may contribute to its pathogenesis.
