Hematology
Contributions
Type: Publication Only
Background
Iron homeostasis is dysregulated in primary myelofibrosis (PMF), given the high prevalence of anemia, need for red blood cell (RBC) transfusions, and disease-associated inflammatory state. Recent data published by Pardanani et al. (Am. J. Hematol. 88:312–316, 2013) show that hepcidin levels in patients diagnosed with Myelofibrosis (MF) are considerably elevated (MF patients median 156,3 pg/mL, Healthy patients median 13,5 pg/mL) and are strongly correlated with serum ferritin levels and even increased hepcidin levels were associated with shortened survival.
Aims
To appraise serum hepcidin levels in MF patients treated with JAK inhibitors.
Methods
Plasma hepcidin levels were measured in six patients diagnosed with Myelofibrosis, all of them with intermediate-2 or high IPSS-risk, were on treatment with a JAK-1/JAK-2 inhibitor (Ruxolitinib) at least 6 months prior. All patients with transfusion dependence were receiving iron chelation therapy (except patient 1). The methodology for hepcidin measurement was DRG Hepcidin-25 (bioactive) ELISA kit. It is a solid phase enzyme-linked immunosorbent assay (ELISA) based on the principle of competitive binding (DRG Instruments GmbH, Germany). The microtiter wells are coated with a monoclonal (mouse) antibody directed towards an antigenic site of the hepcidin-25 molecule. Endogenous hepcidin-25 of a sample competes with a hepcidin-25-biotin conjugate for binding to the coated antibody. After incubation, the unbound conjugate is washed off and a streptavidin-peroxidase enzyme complex is added to each well. After incubation, unbound enzyme complex is washed off and substrate solution is added. The blue color development is stopped after a short incubation time, turning the color from blue to yellow. The absorbance of each well is determined at 450 nm with a microtiter plate reader. The intensity of color developed is reverse proportional to the concentration of hepcidin in the sample. A standard curve is calculated using a 4PL (4 Parameter Logistics) curve fit. The concentrations of the samples can be read directly from this standard curve. Data from serum ferritin level, transferrin saturation index and transfusion dependence was collected from their medical records.
Results
The results are summarized in the table.
| Serum Hepcidin | Serum Ferritin | Transferrin Saturation Index | Transfusion requirements |
Patient 1+ | 94 ng/mL | 8825 ng/mL | 107.2 % | 4 units / month |
Patient 2 | 12 ng/mL | 223 ng/mL | 45.4 % |
|
Patient 3 | 40 ng/mL | 1235 ng/mL | 95.6 % | 3 units / month |
Patient 4 | 24 ng/mL | 116 ng/mL | 20.9 % |
|
Patient 5+ | 138 ng/mL | 2091 ng/mL | 60.4 % | 4 units / month |
Patient 6 | 23 ng/mL | 862 ng/mL | 43.9 % |
|
*median serum Hepcidin in control cohort: 9.8 ng/mL.
+Dead.
Summary
In our cohort of patients diagnosed with Myelofibrosis we can observe elevated levels of hepcidin and its direct correlation with serum ferritin levels. In turn, we realize the hepcidin levels represent a negative prognostic factor in survival of these patients. Comparing hepcidin levels observed in our patients with the data published by Pardanani et al we can infer that Ruxolitinib plays a role in lowering serum hepcidin level.
Keyword(s): Hepcidin, Iron overload, Myelofibrosis
Session topic: Publication Only
Type: Publication Only
Background
Iron homeostasis is dysregulated in primary myelofibrosis (PMF), given the high prevalence of anemia, need for red blood cell (RBC) transfusions, and disease-associated inflammatory state. Recent data published by Pardanani et al. (Am. J. Hematol. 88:312–316, 2013) show that hepcidin levels in patients diagnosed with Myelofibrosis (MF) are considerably elevated (MF patients median 156,3 pg/mL, Healthy patients median 13,5 pg/mL) and are strongly correlated with serum ferritin levels and even increased hepcidin levels were associated with shortened survival.
Aims
To appraise serum hepcidin levels in MF patients treated with JAK inhibitors.
Methods
Plasma hepcidin levels were measured in six patients diagnosed with Myelofibrosis, all of them with intermediate-2 or high IPSS-risk, were on treatment with a JAK-1/JAK-2 inhibitor (Ruxolitinib) at least 6 months prior. All patients with transfusion dependence were receiving iron chelation therapy (except patient 1). The methodology for hepcidin measurement was DRG Hepcidin-25 (bioactive) ELISA kit. It is a solid phase enzyme-linked immunosorbent assay (ELISA) based on the principle of competitive binding (DRG Instruments GmbH, Germany). The microtiter wells are coated with a monoclonal (mouse) antibody directed towards an antigenic site of the hepcidin-25 molecule. Endogenous hepcidin-25 of a sample competes with a hepcidin-25-biotin conjugate for binding to the coated antibody. After incubation, the unbound conjugate is washed off and a streptavidin-peroxidase enzyme complex is added to each well. After incubation, unbound enzyme complex is washed off and substrate solution is added. The blue color development is stopped after a short incubation time, turning the color from blue to yellow. The absorbance of each well is determined at 450 nm with a microtiter plate reader. The intensity of color developed is reverse proportional to the concentration of hepcidin in the sample. A standard curve is calculated using a 4PL (4 Parameter Logistics) curve fit. The concentrations of the samples can be read directly from this standard curve. Data from serum ferritin level, transferrin saturation index and transfusion dependence was collected from their medical records.
Results
The results are summarized in the table.
| Serum Hepcidin | Serum Ferritin | Transferrin Saturation Index | Transfusion requirements |
Patient 1+ | 94 ng/mL | 8825 ng/mL | 107.2 % | 4 units / month |
Patient 2 | 12 ng/mL | 223 ng/mL | 45.4 % |
|
Patient 3 | 40 ng/mL | 1235 ng/mL | 95.6 % | 3 units / month |
Patient 4 | 24 ng/mL | 116 ng/mL | 20.9 % |
|
Patient 5+ | 138 ng/mL | 2091 ng/mL | 60.4 % | 4 units / month |
Patient 6 | 23 ng/mL | 862 ng/mL | 43.9 % |
|
*median serum Hepcidin in control cohort: 9.8 ng/mL.
+Dead.
Summary
In our cohort of patients diagnosed with Myelofibrosis we can observe elevated levels of hepcidin and its direct correlation with serum ferritin levels. In turn, we realize the hepcidin levels represent a negative prognostic factor in survival of these patients. Comparing hepcidin levels observed in our patients with the data published by Pardanani et al we can infer that Ruxolitinib plays a role in lowering serum hepcidin level.
Keyword(s): Hepcidin, Iron overload, Myelofibrosis
Session topic: Publication Only