Contributions
Abstract: PB2260
Type: Publication Only
Background
Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases involved in remodeling of the extracellular matrix. MMPs have multiple roles in cancer cell proliferation and inflammation, including myeloproliferative neoplasm (MPN).
Aims
The goal of our study is to explore the MMPs level in essential thrombocythemia (ET) and primary myelofibrosis (PMF), during inflammation.
Methods
Using zymography, we analyzed expression of MMP2,3,9 and 13 in plasma of healthy controls (n=5), ET (n=17) and PMF (n=17), according to JAK2V617F, calreticulin (CALR) and thrombopoietin receptor (MPL) mutation status by DNA sequencing. In addition, we determined the level of MMP2 in JAK2V617F mutated human erythroleukemia cell line (HEL) treated with pro-inflammatory IL6 and anti-inflammatory IL10. Also, MMP2 levels were determined in peripherial blood mesenchymal stromal cells of healthy controls, with or without mononuclear cells of PMF origin, treated with IL6 and JAK1/2 inhibitor Ruxolitinib. We divide ET and PMF patients per three groups: JAK2V617F positive (JAK2+, n=6), CALR positive (CALR+, n=6), and triple (JAK2V617F / CALR / MPL) negative (n=5).
Results
The levels of MMP3 and MMP13 were significantly higher in all three groups of PMF patients (p<0.001) compared to control and moreover the level of MMP3 and MMP2 were significantly lower (p<0.01) in triple negative ET/PMF compared to JAK2+ and CALR+ ET/PMF patients. The MMP9 was significantly higher in triple negative and JAK2+ patients with PMF compared to control (p<0.01). The level of MMP2 was significantly higher and in ET JAK2+ and CALR+ compared to control (p<0.001). The level of MMP3 was statistically lower in ET triple negative and JAK2+ patients compared to healthy controls (p<0.01), in opposite to MMP9 levels. The MMP13 levels were significantly higher and in ET JAK2+ and CALR+ compared to control (p<0.001) and triple negative ET (p<0.01). IL-6 induced expression of MMP2 in HEL cells after 6 hours (p<0.01), but not IL-10. IL-6 induced expression of MMP2 in mesenchymal stromal cells (p<0.01), while Ruxolitinib inhibited MMP2 in PMF mononuclear cells in co-culture with mesenchymal stromal cells after 48 hours.
Conclusion
The examined MMP levels were increased in plasma of ET and PMF patients, with predominance in JAK2+ and CALR+ patients. Inflammation increased MMP2 levels demonstrating JAK1/2 dependence. These observed MMP result may be a potential diagnostic marker of MPN.
Session topic: 15. Myeloproliferative neoplasms – Biology & Translational Research
Keyword(s): inflammation, MMP, Myeloproliferative disorder
Abstract: PB2260
Type: Publication Only
Background
Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases involved in remodeling of the extracellular matrix. MMPs have multiple roles in cancer cell proliferation and inflammation, including myeloproliferative neoplasm (MPN).
Aims
The goal of our study is to explore the MMPs level in essential thrombocythemia (ET) and primary myelofibrosis (PMF), during inflammation.
Methods
Using zymography, we analyzed expression of MMP2,3,9 and 13 in plasma of healthy controls (n=5), ET (n=17) and PMF (n=17), according to JAK2V617F, calreticulin (CALR) and thrombopoietin receptor (MPL) mutation status by DNA sequencing. In addition, we determined the level of MMP2 in JAK2V617F mutated human erythroleukemia cell line (HEL) treated with pro-inflammatory IL6 and anti-inflammatory IL10. Also, MMP2 levels were determined in peripherial blood mesenchymal stromal cells of healthy controls, with or without mononuclear cells of PMF origin, treated with IL6 and JAK1/2 inhibitor Ruxolitinib. We divide ET and PMF patients per three groups: JAK2V617F positive (JAK2+, n=6), CALR positive (CALR+, n=6), and triple (JAK2V617F / CALR / MPL) negative (n=5).
Results
The levels of MMP3 and MMP13 were significantly higher in all three groups of PMF patients (p<0.001) compared to control and moreover the level of MMP3 and MMP2 were significantly lower (p<0.01) in triple negative ET/PMF compared to JAK2+ and CALR+ ET/PMF patients. The MMP9 was significantly higher in triple negative and JAK2+ patients with PMF compared to control (p<0.01). The level of MMP2 was significantly higher and in ET JAK2+ and CALR+ compared to control (p<0.001). The level of MMP3 was statistically lower in ET triple negative and JAK2+ patients compared to healthy controls (p<0.01), in opposite to MMP9 levels. The MMP13 levels were significantly higher and in ET JAK2+ and CALR+ compared to control (p<0.001) and triple negative ET (p<0.01). IL-6 induced expression of MMP2 in HEL cells after 6 hours (p<0.01), but not IL-10. IL-6 induced expression of MMP2 in mesenchymal stromal cells (p<0.01), while Ruxolitinib inhibited MMP2 in PMF mononuclear cells in co-culture with mesenchymal stromal cells after 48 hours.
Conclusion
The examined MMP levels were increased in plasma of ET and PMF patients, with predominance in JAK2+ and CALR+ patients. Inflammation increased MMP2 levels demonstrating JAK1/2 dependence. These observed MMP result may be a potential diagnostic marker of MPN.
Session topic: 15. Myeloproliferative neoplasms – Biology & Translational Research
Keyword(s): inflammation, MMP, Myeloproliferative disorder