Higher Hepcidin Levels Research Articles

Acquired Refractory Iron Deficiency Anemia.

Anemia is a global health problem affecting one-third of the world population, and half of the cases are due to iron deficiency (ID). Iron deficiency anemia (IDA) is the leading cause of disability in several countries. Although multiple mechanisms may coexist, ID and IDA causes can be classified as i) insufficient iron intake for the body requirement, ii) reduced absorption, and iii) blood losses. Oral iron represents the mainstay of IDA treatment. IDA is defined as “refractory” when the hematologic response after 4 to 6 weeks of treatment with oral iron (an increase of >=1 g/dL of Hb) is absent. The cause of iron-refractory anemia is usually acquired and frequently related to gastrointestinal pathologies, although a rare genetic form called iron-refractory iron deficiency anemia (IRIDA) exists. In some pathological circumstances, either genetic or acquired, hepcidin increases, limiting the absorption in the gut, remobilization, and recycling of iron, thereby reducing iron plasma levels. Indeed, conditions with high hepcidin levels are often under-recognized as iron refractory, leading to inappropriate and unsuccessful treatments. This review provides an overview of the iron refractory anemia underlying conditions, from gastrointestinal pathologies to hepcidin dysregulation and iatrogenic or provoked conditions, and the specific diagnostic and treatment approach.

Plasma Hepcidin Level in Hemodialysis Patients and its Relationship to Anemia Therapy and Dialysis Efficiency

Background: Hepcidin is a small peptide produced mainly by hepatocytes, macrophages and other cells. In addition to its antimicrobial effects, it is the primary regulator of iron metabolism as it controls both dietary iron absorption from the duodenum and the release of that iron by reticuloendothelial cells. Objective: Study of plasma hepcidin level and its relationship to the parameters of iron status, anemia therapy, and parameters of dialysis efficiency in hemodialysis patient. Patients and Methods: 106 subjects were enrolled in our study. They were divided into two groups: Group 1 (control group) included10 healthy subjects. Group 2 included96 end-stage renal disease (ESRD) patients on regular hemodialysis; they were subdivided into two groups according to the median plasma hepcidin level. full history, physical examination and detailed analysis of medical records to elucidate the cause of renal failure if present and duration of hemodialysis, estimation of glomerular filtration rate (eGFR) and calculating Kt/V. Laboratory investigations includingCBC, liver function tests, kidney function tests, serum iron, serum ferritin, total iron binding capacity (TIBC), transferrin saturation and plasma hepcidin level were measured. In addition to Pelvi-abdominal ultrasound was done. Results: Plasma hepcidin levels were higher in maintenance hemodialysis patients than in healthy control subjects. Also, there was a significant and independent positive correlation between hepcidin and both serum ferritin levels and KT/V. While there was significant negative correlation between serum hepcidin levels & EPO dose. Conclusion: High plasma hepcidin levels in HD patients depending on the magnitude of the inflammatory process and on rhEPO doses. Close interaction between hematological indices, iron status and plasma hepcidin levels.

Low anticoagulant heparin-iron complex targeting inhibition of hepcidin ameliorates anemia of chronic disease in rodents

Hepcidin is the only known hormone negatively regulates systemic iron availability, its excess contributes to anemia of chronic disease (ACD).Heparin has been shown to be an efficient hepcidin inhibitor both in vitro and in vivo, but its powerful anticoagulant activity limits this therapeutic application. To this end, heparin-iron complex was prepared by electrostatic interaction and/or coordination between heparin and dihydroxy iron solution ([Fe(OH)2]+) under the condition of ultrasonic assisted. We assessed the anticoagulant activity of heparin-iron in vitro and vivo by sheep plasma, chromogenic substrate method and tail-bleeding in mice, respectively. Anti-hepcidin effect of heparin-iron was detected in HepG2 cell and LPS induced acute inflammation mice by qRT-PCR and ELISA. Turpentine-induced anemia mice were established to evaluate the effect of heparin-iron in ACD. Mice were treated with heparin-iron for 4 weeks. The results indicated that heparin-iron has significantly reduced anticoagulant activity in vitro and in vivo, strongly decreases hepcidin mRNA and IL-6 induced high level of secreted hepcidin in HepG2 cell. Heparin-iron was also found to cause a reduction on hepcidin expression through BMP/SMAD and JAK/STAT3 pathways in LPS induced acute inflammation model in mice. In ACD mice, heparin-iron could lower elevated serum hepcidin and improve anemia. These findings demonstrated low anticoagulant heparin-iron has potential applications for the treatment of ACD with high hepcidin levels.

Molecular analysis of homeostatic iron regulator, transmembrane protease serine-6, and BTB domain-containing protein-9 variants and iron parameters in blood donors

Genetic variants associated with iron homeostasis have been identified, but their association with iron-related indices and variables among different ethnic populations remains controversial. We aimed to explore the genotype frequency and allelic distribution of three iron-metabolism related variants in homeostatic iron regulator gene (HFE; rs1800562 G/A), transmembrane protease, Serine-6 gene (TMPRSS6; rs855791 A/G), and BTB domain-containing protein-9 gene (BTBD9; rs9357271 C/T) among a sample of the Middle Eastern blood donors and to detect the association of these variants on blood indices, and serum hepcidin/ferritin levels. Real-Time TaqMan genotyping assay for the specified variants was applied for 197 unrelated blood donors. Complete blood picture and serum hepcidin/ferritin levels were assessed. All participants were carriers of rs1800562*G/G genotype for HFE. The frequency of A/A and A/G genotypes of TMPRSS6 rs855791 variant was 55% and 45%, and for C/C, C/T, and T/T of BTBD9 rs9357271, were 15%, 43%, and 42%, respectively. Minor allele frequencies of rs855791*G and rs9357271*C were 0.23 and 0.37. The GGC genotype combination (for HFE/TMPRSS6/BTBD9, respectively) was more frequent in male participants. Higher serum hepcidin and hepcidin/ferritin ratio were observed in TMPRSS6 (A/G) carriers. While subjects with BTBD9 C/T and TT genotypes had lower serum ferritin values and higher levels of hepcidin and hepcidin/ferritin ratio compared with C/C genotype. No significant associations were found with any other blood parameters.In conclusion, TMPRSS6 rs855791 (A/G) and BTBD9 rs9357271 (C/T) variants were prevalent in the present blood donor population and may influence the serum hepcidin and/or ferritin levels.

Verapamil administration alleviates microcytosis and tissue accumulation after chronic aluminum exposure in rats

Introduction/Objective. Research has demonstrated the toxicant potential of aluminum, but no therapeutic options have been suggested. The aim of the study was to investigate the extent of the aluminuminduced toxicity, evaluated by hematological/biochemical disarrangements, hepcidin concentration and tissue accumulation after chronic aluminum exposure and to determine possible protection with Ca2+-channel blockage, verapamil. Methods. Experimental animals (36 rats) were treated with different doses of AlCl3 during eight weeks and after that their blood and tissues were analyzed. Results. The significant differences, regardless of the aluminum dose administered, were documented in all evaluated hematological (p < 0.001) and biochemical parameters (p < 0.001), as well as in aluminum tissue deposition in liver, kidneys and testicles (p < 0.001), respectively. After verapamil administration, a significant improvement in some hematological and biochemical parameters was demonstrated, p < 0.001, as well as the attenuation of aluminum deposits in liver and testes, p < 0.001. Evaluated parameters of inflammation and kidney deposition did not show significant change after verapamil application. Conclusion. The findings indicate that chronic AlCl3 intoxication, regardless of the dose, results in the microcytic anemia associated with high hepcidin levels, numerous biochemical abnormalities and significant aluminum deposition in liver, kidney and testes and that these effects may be attenuated by verapamil administration. Overall, the results emphasize the significance of calcium homeostasis preservation in chronic aluminum exposure and propose possible therapeutic option.

The Hepcidin and 25-OH-Vitamin D Levels in Obese Children as a Potential Mediator of the Iron Status.

Obesity is associated with the risk factors such as iron and vitamin D deficiencies. Increased risk of iron deficiency generally correlates with the high levels of serum hepcidin in obese children. Vitamin D deficiency was also linked to an increase in serum hepcidin levels. We aimed to compare iron parameters and investigate the hepcidin levels in obese and non-obese children. This study included 83 children and adolescents including obese (n = 35) and non-obese (n = 48). Laboratory values including serum iron levels, total iron-binding capacity, percentage of transferrin saturation, ferritin, reticulocyte parameters and high sensitivity C-reactive protein (hsCRP), hepcidin, 25-OH-Vitamin D were measured. Average levels of hepcidin, hsCRP, and ferritin were found to be similar in both study groups. Serum iron levels, total iron-binding capacity, percentage of transferrin saturation, and 25-OH-vitamin D levels were significantly lower in the obese group. There was no statistically significant difference between hepcidin and 25-OH-vi-tamin D levels. Average hepcidin levels were detected to be similar in both groups (p = 0.580) whereas, 25-OH-vi-tamin D levels were significantly lower in the obese group (p < 0.001). A statistically negative correlation was observed between average BMI (body mass index) and serum iron level (r = -0.476; p < 0.001), BMI and transferring saturation (r = -0.467; p < 0.001), and BMI and 25-OH-vitamin D levels (r = -0.474; p < 0.001). Hence, no statistically significant relation was detected between hepcidin and 25-OH-vitamin D levels (r = 0.233; p = 0.084). Being female, vitamin D deficiency, and IRF (%) (Immature Reticulocyte Fraction) were found as independent risk factors for BMI increase due to logistic regression analyses. In conclusion, observed statistical associations and correlations do not prove a causal relationship between the hepcidin levels and iron deficiency but vitamin D deficiency seems likely to cause high BMI levels or in contrast, obesity may cause vitamin D deficiency in the children. No association was found between hepcidin, ferritin, and hsCRP levels with obesity in children. However, vitamin D deficiency was detected to cause a 5.3-fold increase in BMI levels. We suggest that there may be different mechanisms in obesity-related metabolic and hematological events. One can also envision that there is not enough time for the chronic inflammation processes to develop during childhood as opposed to those frequently seen in adult obese individuals.

Severity of clinical manifestation, and prognosis of patients with myelofibrosis

Introduction: Primary myelofibrosis (PMF) is a myeloproliferative disorder characterized by bone marrow fibrosis and ineffective extramedullary hematopoiesis, which presents with anemia, constitutional symptoms, and excessive splenomegaly. A number of factors are involved in the pathogenesis of the anemia in PMF, including impaired iron metabolism regulation. It was found that higher levels of the key regulator of iron metabolism - the peptide hormone hepcidin in patients with PMF, are associated with the severity of the anemia, blood transfusion dependence and decreased overall survival. Aim: The aim of the study was to analyze the serum levels of hepcidin in patients with PMF and its impact on the clinical course, prognosis, and outcome of the disease. Materials and Methods : A total of 68 patients with PMF and 12 healthy controls were analyzed. Serum hepcidin levels were measured by ELISA. The results were statistically analyzed by dispersion, comparison, and correlation methods. Results: Then mean hepcidin levels in patients with PMF were statistically significantly higher compared to healthy controls. (99.05 ng/mL; 20.57 ng/mL; F = 7.95; p = 0.006). High levels of hepcidin correlated with high risk according to DIPSS (p = 0.046), carrier of JakV617F mutation (p = 0.022), fibrotic phase according to WHO 2016 (p = 0.062), and the number of blood transfusions per month (p = 0.005). Higher hepcidin levels were not relevant to overall survival. Conclusion: Hepcidin is a biological marker the monitoring of which in the course of MF would help for a more accurate clinical and prognostic assessment of the disease.

The Dysregulation of Hepcidin-Ferroportin Axis in Childhood Acute Lymphoblastic Leukemia Survivors after Completion of Chemotherapy

Background The hepcidin-ferroportin axis is known to play the essential role on iron homeostasis in human. The binding of hepcidin to ferroportin (FPN) on the membranes of iron-exporting cells induces the ubiquitination of FPN and thereby decreases the delivery of iron to plasma. Low hepcidin with high FPN was well established in patients with ineffective erythropoiesis, e.g. beta-thalassemia major and myelodysplastic syndrome. On the other hand, studies showed increased hepcidin accompanied with decreased FPN in patients with some cancers. Further, anemia in multiple myeloma and Hodgkin's lymphoma were reported to be associated with high hepcidin levels (Blood 2008;112:4292). Here, we investigated the function of the hepcidin-ferroportin axis in childhood acute lymphoblastic leukemia (ALL) survivors who completed chemotherapy. Methods Thirty-three children, aged ≤18 years with ALL and completing TPOG-ALL-2013 protocol before June 30, 2020, were enrolled (ALL-off Gr). According to the treatment protocol, they had been classified as standard-risk (SR) in 20, high-risk (HR) in 12 and very-high-risk (VHR) in 1, respectively. Thirty-eight healthy siblings were assigned as the normal controls (normal Gr) after exclusion of acute illness and iron deficiency. Twenty newly-diagnosed ALL children (ALL-Dx Gr) who did not have sepsis at diagnosis (Dx), and 13 patients with beta-thalassemia major (TM Gr) who received regular transfusion were also enrolled. Serum iron, total iron-binding capacity, ferritin, hepcidin and FPN were collected at Dx in ALL-Dx Gr and at 6-month follow-up after completion of chemotherapy in ALL-off Gr. Selected biochemical assays were checked in normal Gr and TM Gr on the day of routine RBC transfusion. Hepcidin and FPN levels of peripheral blood were measured using ELISA kits (DRG, EIA-5782 for hepcidin &amp; Elabscience, E-EL-H2355 for FPN, respectively). Results The clinical features and biochemical data of all patients are shown in Table 1. The hepcidin and FPN of normal Gr were comparable with those of reference papers. The low hepcidin, high FPN and relatively low hepcidin/ferritin of TM Gr were in consistent with those of published reports. There were no significant differences of age (P =.243) and gender (P =.899) between ALL-off Gr and normal Gr. The ferritin and transferrin saturation between ALL-Dx and ALL-off Grs showed statistical difference (P &amp;lt;.001 and P =.02, respectively). In figure 1, the hepcidin and FPN levels of ALL-Dx Gr were significantly higher than those of the normal Gr (P &amp;lt;.001). The trends remained but did not reach statistical significances between ALL-off and normal Grs. None of ALL-off Gr had mutation of HFE C282Y/H63D, HJV C119F or SLC40A1 C326S. Further, in ALL-off Gr, 6 patients had abnormal ferritin levels (&amp;gt;300 ng/mL; median, 871.55; range, 351.6-959.0)(high-ferritin). MRI scans of liver and heart from the 6 high-ferritin patients did not show any evidence of iron overload. In Figure 2, hepcidin levels were significantly higher in ALL-off patients with high-ferritin or receiving cumulative RBC transfusions &amp;gt;10 units. And ALL-off patients with high-ferritin, RBC transfusions &amp;gt;10 units or receiving HR/VHR treatment regimens had significantly higher FPN levels. Multivariate analysis demonstrated only high-ferritin predicted significantly higher hepcidin (P =.02) and FPN (P &amp;lt;.001) levels in ALL-off Gr. Conclusions The dysregulation of hepcidin-ferroportin axis showed high hepcidin and uncontrolled FPN hyperactivity at Dx of childhood ALL which may point to the impaired down-regulation of FPN by hepcidin. Our findings imply the impaired mechanism may not be fully recovered in ALL survivors after completing chemotherapy, especially those with ferritin &amp;gt;300 ng/mL, HR/VHR groups or receiving higher RBC transfusions. The routine monitoring of the axis function could facilitates the early detection of iron overload in liver and heart. Disclosures No relevant conflicts of interest to declare.

Administration of KER-047, a Novel ALK2 Inhibitor, Elicited Robust and Sustained Increases in Serum Iron in Healthy Participants

Background: Hepcidin is the master regulator of iron homeostasis. Its expression is controlled, in part, by signaling through TGF-β receptors including activin receptor-like kinase-2 (ALK2). As part of a negative feedback loop, high hepcidin levels increase expression of matriptase-2, which cleaves the ALK2 co-receptor hemojuvelin from the cell surface and suppresses ALK2 signaling. Dysregulation of the negative feedback system results in high hepcidin, which mobilizes iron from storage tissues and leads to insufficient iron for red blood cell production in the bone marrow, resulting in anemia. KER-047 is a selective ALK2 inhibitor that has the potential to normalize ALK2 signaling and the downstream sequalae that results in anemia as a result of elevated hepcidin. Aims: The objective of this Phase 1 study was to evaluate the safety, tolerability, pharmacokinetics, and pharmacodynamic effects of single and multiple ascending dose levels of KER-047 in healthy participants. Methods: In Part 1, two formulations of KER-047 were evaluated in single ascending oral doses ranging from 1 mg to 300 mg of a capsule formulation, and 30 mg to 450 mg of a liquid formulation or placebo. In Part 2, the liquid formulation was evaluated in multiple ascending doses (50 mg, 100 mg, 200 mg and 350 mg) of KER-047 or placebo, administered daily for up to 7 days. Endpoints included adverse events (AEs) and pharmacokinetic and pharmacodynamic parameters. Informed consent was obtained from all participants prior to enrollment in the study. Results: Part 1 enrolled 80 participants; 60 received a single dose of KER-047 and 20 received placebo. Part 2 enrolled 41 participants; 32 received KER-047 and 9 received placebo daily for up to 7 days. There were no serious adverse events in either part of the study. In Part 1, 3 (9.4%) participants discontinued the study; none discontinued due to AEs. In Part 2, 3 (9.4%) participants administered KER-047 and 1 (11.1%) placebo discontinued the study; of these, 2 participants discontinued due to an AE (one in the 200 mg group and one on placebo). In Part 2, 1 of 8 (12.5%) administered 200 mg and 4 of 8 (50%) participants administered 350 mg discontinued study drug due to AEs. The majority of AEs observed in participants administered KER-047 were mild or moderate in severity; severe AEs were reported only at the 350 mg multiple dose in 1 of 8 (12.5%) participants. AEs reported in ≥2 of participants administered KER-047 and higher than placebo were: headache, nausea, vomiting, diarrhea, gastroenteritis, chills, pyrexia, myalgia, decreased appetite, lymphopenia, neutropenia, and liver enzyme increases. Decreases in lymphocyte and neutrophil counts were observed at 200 mg and 350 mg. Mean KER-047 AUC and Cmax increased linearly, with greater than dose-proportional increases observed across multiple doses from 50-200 mg. Half-life values ranged from approximately 10 to 15 hours. Administration of KER-047 elicited rapid, robust and sustained dose-related increases in serum iron and transferrin saturation. Increases in serum iron and transferrin saturation were observed beginning on Day 2 after single doses and were sustained after multiple doses. Decreases in ferritin were also observed. Decreases in serum hepcidin post-dose were also observed in all the multiple dose cohort in which changes were evaluated. Reticulocyte hemoglobin content increased starting on Day 3 post-dosing indicating increased iron availability in the bone marrow. Summary: In healthy participants, administration of KER-047 elicited rapid, robust and sustained dose-related increases in serum iron and transferrin saturation that were associated with decreases in ferritin and hepcidin. The observed decrease in ferritin and hepcidin coupled with increases in reticulocyte hemoglobin content are indicative of increased iron mobilization, resulting in increased iron incorporation into hemoglobin. The tolerability profile of KER-047 in healthy participants has been characterized in this Phase 1 study. KER-047's unique pharmacologic effect on hepcidin and iron mobilization has the potential to treat anemia that results from elevated hepcidin. Table Disclosures Ordonez: Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company; Ackea Therapeutics: Ended employment in the past 24 months. Ben:Nucleus Network: Current Employment. Barger:Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company. Serino:BioBridges: Ended employment in the past 24 months; Keros Therapeutics: Current Employment. Tseng:Mitobridge: Current equity holder in private company; Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company, Patents &amp; Royalties. Rovaldi:Keros Therapeutics: Consultancy. Lachey:Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company. Seehra:Keros Therapeutics: Current Employment, Current equity holder in publicly-traded company.

Importance of Hepcidin in the Etiopathogenesis of Anemia in Inflammatory Bowel Disease.

Anemia is the most common extraintestinal systemic complication of inflammatory bowel disease. Iron deficiency anemia and anemia of chronic disease are among the most frequent types. Intestinal iron absorption is controlled by the activity of ferroportin. Cells with high expression of ferroportin include enterocytes, and also macrophages and hepatocytes. Iron homeostasis is controlled by the hepcidin-ferroportin axis. Hepcidin is a central regulator of iron metabolism and can also serve as a marker of systemic inflammation. During systemic inflammatory response, the synthesis of hepcidin increases, and hepcidin binds to ferroportin and inhibits its activity. Thus, iron is not absorbed from the bowel into the circulation and also remains sequestered in macrophages. Conversely, hepcidin synthesis is suppressed during conditions requiring increased iron intake for enhanced erythropoiesis, such as iron deficiency anemia or hypoxia. Here, ferroportin is not blocked, and iron is actively absorbed into the bloodstream and also released from the stores. Production of hepcidin is influenced by the status of total body iron stores, systemic inflammatory activity and erythropoietic activity. Oral iron therapy is limited in inflammatory bowel diseases due to ongoing gastrointestinal inflammation. It is less effective and may worsen the underlying disease. Therefore, the choice between oral and parenteral iron therapy must be made with caution. Oral iron would be ineffective at high hepcidin levels due to concurrent ferroportin blockage. Contrarily, low levels of hepcidin indicate that oral iron therapy should be successful. An understanding of hepcidin can help in understanding the body's reaction to iron depletion during the inflammatory process.

Associations of NT-proBNP and hepcidin levels with clinical and laboratory parameters in patients with heart failure with various severity of left ventricular systolic dysfunction

Data on hepcidin levels in patients with heart failure (HF) are contradictory and do not make clear its contribution to the progression of multiple organ failure. There remain a number of issues about the prognostic significance of the N-terminal pro-brain natriuretic peptide (NT-proBNP) in HF with preserved ejection fraction (EF). The authors suggested the relationships between these markers in decompensated HF, as well as their associations with other clinical and laboratory parameters.Aim. To identify the association of NT-proBNP and hepcidin levels with clinical and laboratory parameters in patients with HF with various severity of left ventricular (LV) systolic dysfunction.Material and methods. The study included 68 patients (29 women, 39 men; mean age — 72,3±11,7 years) hospitalized due to decompensated HF. Patients were divided into three groups: reduced (HFrEF) (n=20), mid-range (HFmrEF) (n=23), and preserved EF (HFpEF) (n=24). Upon admission, along with standard diagnostic tests, all patients were examined for NT-proBNP and hepcidin levels by enzyme-linked immunosorbent assay. Statistical processing was carried out using the software package Statistica 8.0.Results. NT-proBNP levels in the entire sample was 315,9 [129,9; 576,1] pg/ml. Significantly higher concentrations of NT-proBNP were found in patients with lower EF: 433,05 (346,8-892,6) pg/ml for HFrEF, 289,97 (185,9-345,3) pg/ml for HFmrEF pg/ml and 214,98 (207,37-562,31) pg/ ml for HFpEF (p&lt;0,05). At the same time, hepcidin levels in the HFrEF group (31,63 ng/ml [22,0; 71,6]) was significantly higher than in the HFmrEF (23,89 ng/ml [21,1; 27,9]) (p&lt;0,05) and HFpEF (26,91 ng/ml [18,6; 31,1]) (p&lt;0,05). In HFpEF patients, there was a correlation of hepcidin level with body mass index (r=0,47, p&lt;0,05) and chronic obstructive airway diseases (r=0,44, p&lt;0,05). A correlation of hepcidin level with cardiac arrhythmias (r=0,61, p&lt;0,05) was revealed in the HFmrEF group. In the HFrEF group, there were correlations of a significantly increased level of NT-proBNP (median — 433,05; 95% confidence interval: 346,8-892,6) with indicators of disease severity and multiple organ dysfunction: decrease in systolic blood pressure, cardiorenal syndrome, decrease in hemoglobin level and mean corpuscular hemoglobin concentration, characteristic of iron-deficiency anemia.Conclusion. Patients with lower EF showed higher NT-proBNP values and a trend towards higher hepcidin levels. Relationships of hepcidin and NT-proBNP levels with following clinical parameters were found: body mass index, presence of obstructive airway diseases, cardiac arrhythmias, as well as low cardiac output syndrome, cardiorenal syndrome and anemia.

Serum Hepcidin Level in Patients with Acute Lymphoblastic Leukemia (ALL) during The Treatment Phase: Their Effects on Erythropoisis Activity and Iron Reserves

Hepcidin levels increased significantly with increased of iron stores in early phase of acute leukemia patients when erithropoisis pressed by blast cells in bone marrow, then decreased significantly with acute leukemia remission. The study aimed to determine role of hepcidin in activity of erythropoisis and serum iron reserves in acute lymphoblastic leukemia (ALL) patients during phase of therapy. This study used an observational analytic design. Serum hepcidin examination used ELISA methods. Erythropoisis activity was determined by complete blood and reticulocyte percentage. Iron reserves was determined by serum iron and ferritin levels by ECLIA. Result shows a total of 48 patient subjects were divided into groups of induction, consolidation and maintenance phase with an average age of 6.81 years (induction), 9.7 years (consolidation) and 7.8 years (maintenance). In the normality test with Shapiro-Wilk data showed abnormal distribution (p <0.05). Analysis by the Kruskal-Wallis test showed there were differences between the three treatment phases in the examination of hemoglobin, reticulocytes, serum iron, ferritin and hepcidin (p <0.05). In statistical analysis with Spearman's rank correlation shows there is a significant correlation between hemoglobin with ferritin (r = -0.416, p = 0.003), hemoglobin with hepcidin (r = -0.305, p = 0.035), reticulocytes with hepcidin (r = -0.496), p = 0.000) and serum iron with hepcidin (r = -0.302, p = 0.037). We concluded that the higher levels of hepcidin indicate lower levels of hemoglobin, reticulocytes and serum iron in patients with acute lymphoblastic leukemia during treatment phase.

Hepcidin and ferritin levels in restless legs syndrome: a case\u2013control study

The association between restless legs syndrome (RLS) and iron homeostasis remains unclear. We compared serum hepcidin and ferritin levels in patients with RLS and controls, and assessed their relationships with RLS phenotype, drug intake, and history of augmentation syndrome. 102 drug-free RLS patients (age 58.9 [24.5–77.2], 63 females) and 73 controls (age 56.8 [23.46–76.6], 45 females) underwent a polysomnography recording. Hepcidin levels were quantified by ELISA. 34 RLS patients had a second assessment after starting dopaminergic drugs. Ferritin level was low (< 50 µg/l) in 14.7% of patients and 25% of controls, with no between-group differences in the mean values. Hepcidin levels were higher in patients even after adjustment for confounding factors, and excluding participants with low ferritin levels. Ferritin and hepcidin levels were comparable before and after treatment, and between patients with (n = 17) and without history of augmentation. Ferritin and hepcidin levels correlated with age, body mass index, and periodic leg movements. Higher hepcidin levels were associated with older age, older age at RLS onset, less daytime sleepiness and familial RLS. In conclusion, serum hepcidin levels but not ferritin were higher in RLS patients regardless of treatment and history of augmentation. Serum hepcidin may be a more relevant biomarker of RLS than ferritin.

Pathogenesis of anemia in pregnant women with gestational diabetes mellitus

The aim of the research was to establish the role of inflammation mediators and iron metabolism in the pathogenesis of various types of anemic syndrome in pregnant women with gestational diabetes mellitus (GDM). Materials and methods. 32 pregnant patients with GDM were examined; 14 of them had iron deficiency anemia, 18 – anemia of chronic diseases. The enzyme-linked immunosorbent assay was used to determine the concentration of IL-6, hepcidin and a soluble receptor for transferrin in the blood serum of pregnant women, the concentrations of C-reactive protein and transferrin were determined with the method of turbidimetry. Results. It was shown that women with GDM had higher IL-6 level compared to healthy pregnant women, and the concentration of IL-6 did not depend on the type of anemic syndrome. The C-reactive protein concentration was higher in patients with GDM and anemia of chronic diseases than in healthy pregnant women or in pregnant women with iron deficiency anemia. An analysis of iron metabolism markers in pregnant women with GDM established that patients with anemia of chronic diseases had higher hepcidin levels than women with iron deficiency anemia or healthy pregnant women. Conclusions. We established the heterogeneity of the anemic syndrome in pregnancy complicated by GDM. It was confirmed that GDM was accompanied by subclinical inflammation, which was more pronounced in anemia of chronic diseases. The research showed that the mechanism of development of anemia of chronic diseases involving the hepcidin protein was also realized in GDM, characterized by subclinical inflammation. The results indicate the importance of establishing the type of the anemic syndrome in pregnant women with GDM for effective therapeutic follow-up.

Impact of Aeromonas hydrophila and infectious spleen and kidney necrosis virus infections on susceptibility and host immune response in Chinese perch (Siniperca chuatsi)

Co-infections with pathogenic microorganisms are common in aquaculture, resulting in more serious economic losses than single-pathogen infection. Infection of Aeromonas hydrophila (A. hydrophila) often occurs together with infectious spleen and kidney necrosis virus (ISKNV) in Chinese perch (Siniperca chuatsi) culture ponds. In this study, A. hydrophila and ISKNV were inoculated into Chinese perch to mimic individual infection, secondary infection, and mixed infection. The effects of concurrent infections on the susceptibility and the immune response of the host and changes in bacterial and viral load were studied. The results showed relatively complex interaction between ISKNV and A. hydrophila for different infection modes, acting in an antagonistic or synergistic manner. The experimental groups infected with a mixture of ISKNV and A. hydrophila showed higher mortality rate than groups infected with single-pathogen or secondary infection groups, suggesting a synergistic lethal effect of A. hydrophila and ISKNV co-infection. Serious clinical symptoms and obvious histopathological changes were observed in moribund fish under the mixed-infection condition. In addition, obviously higher mortalities were caused by secondary bacterial infections than the number caused by secondary viral infections. ISKNV-primary infection increased the mortality caused by secondary bacterial infections, but A. hydrophila-primary infection did not significantly increase the mortality caused by secondary viral infections. Co-infected fish showed high expression levels of IRF1, Mx, Viperin, Hepcidin, TNFα, and IL-1β mRNAs relative to the levels in healthy fish, which suggested that the co-infection of these two pathogens activated the host immune system and caused host inflammation. These results of infection with A. hydrophila and ISKNV provided the theoretical basis to analyze the pathogenic effects and interaction between pathogens, and could facilitate design of strategies for clinical prevention and control measures of outbreak of fulminant hemorrhagic disease and bacterial sepsis in Chinese perch.

Hepcidin and MELD-XI score as markers of multiple organ failure in patients with heart failure with preserved and reduced ejection fraction

Data on the changes in hepcidin levels in heart failure (HF) patients are contradictory and do not give an answer about its effect on the progression of multiple organ failure. Since the model of end-stage liver disease excluding INR (MELD-XI) reflects the severity of liver and kidney dysfunction, these markers have been suggested to be associated with decompensated HF.Aim. To assess the MELD-XI score and serum hepcidin levels in patients with decompensated HF with different values of left ventricular ejection fraction (EF).Material and methods. The study included 68 patients (29 women, 39 men; mean age 72,3±11,7 years) hospitalized due to decompensated HF. Patients were divided into three groups: reduced (HFrEF) (n=20), mid-range (HFmrEF) (n=23), and preserved EF (HFpEF) (n=24)). Upon admission, along with standard diagnostic tests, all patients were examined for hepcidin-25 levels by enzyme-linked immunosorbent assay. MELD-XI score was calculated. Statistical processing was carried out using the software package Statistica 8.0.Results. Hepcidin levels in the HFrEF group (31,63 ng/ml [22,0; 71,6]) were significantly higher than in the HFmrEF (23,89 ng/ml [21,1; 27,9]) (p&lt;0,05) and HFpEF (26,91 ng/ml [18,6; 31,1]) (p&lt;0,05) groups. In HFpEF, there was a correlation of hepcidin level with body mass index (r=0,47, p&lt;0,05) and chronic obstructive airway diseases (r=0,44, p&lt;0,05). A correlation of hepcidin level with significant cardiac arrhythmias (r=0,61, p&lt;0,05) was revealed in HFmrEF patients. MELD-XI score were significantly increased from 9,44±3,96 for HFpEF and 11,53±3,82 for HFmrEF to 14,3±4,3 for HFrEF (p&lt;0,005). We also revealed correlation of MELD-XI score with hepcidin levels (r=0,3, p&lt;0,05) and EF (r=-0,43, p&lt;0,0003). Patients with a MELD-XI score of &gt;10,4 were more likely to have NYHA class III-IV HF, HFrEF and significantly higher levels of hepcidin (p&lt;0,05 for all) These patients were also more likely to have chronic kidney disease (p&lt;0,05).Conclusion. Hepcidin level and MELD-XI score in patients with decompensated HF are inversely related to left ventricular EF. There is a direct relationship between hepcidin levels and other clinical parameters: body mass index, the presence of chronic obstructive airway diseases and cardiac arrhythmias.

Genetic analysis of TMPRSS6 gene in Saudi female patients with iron deficiency anemia

Objective/backgroundMutations in transmembrane protease serine 6 (TMPRSS6) gene induce high hepcidin level, which causes iron-refractory iron deficiency anemia (IRIDA) by preventing duodenal iron absorption. This study aims to identify the common genetic variations of the TMPRSS6 gene that affect iron levels among Saudi female patients with iron deficiency anemia (IDA). MethodsAll study participants were Saudi females (12–49 years old): 32 patients with IDA, 32 patients with IRIDA, and 34 healthy individuals comprising the control group. Hematological investigations, iron profile, serum hepcidin level, and TMPRSS6 gene transcription were determined. The TMPRSS6 gene was amplified, sequenced, and analyzed among all study participants. ResultsThe mean hepcidin and TMPRSS6 RNA transcription levels in IDA and IRIDA groups were significantly lower than those in the control group. TMPRSS6 gene sequence analysis detected 41 variants: two in the 5′ untranslated region (5′UTR), 17 in introns, and 22 in exons. Thirty-three variants were previously reported in the Single Nucleotide Polymorphism Database, and eight variants were novel; one novel variant was in 5′UTR (g.-2 T > G); five novel variants were detected in exons (p.W73X, p.D479N, p.E523K, p.L674L, and p.I799I). At the time of the sequence analysis of our samples, two variants—p.D479N and p.674L—were novel. However, these variants are present at a very low allele frequency in other populations (L674L, 0.00007761 and D479N, 0.000003980). ConclusionThis is the first study to investigate the genetic variants of TMPRSS6 gene in Saudi female patients with IDA. The generated data will serve as a reference for future studies on IDA in the Arab population.

A hepcidin-based approach for iron therapy in hemodialysis patients: A pilot study.

Hepcidin is a key factor that regulates iron homeostasis. In hemodialysis patients (HD), a high hepcidin level may decrease intestinal iron absorption and reduce the efficacy of Oral iron vs Intravenous iron therapy. Whether the hepcidin level in HD could guide oral iron therapy is unclear. We report a monocentric study on nine "erythropoietin (EPO)-free" patients (without recombinant human EPO [rHU-EPO] for at least 6 months) and normal hepcidin level (<20 ng mL) during the study. After 15 days of washout, oral iron (ferrous sulfate 80 mg/day) was introduced. The primary end point was the hemoglobin response and iron store at 3 months. Nine patients (8 men, 1 woman) with a median age of 62 years (range 42-79) were included. After 1 week of treatment, the median transferrin saturation index increased from 15% (range 6-61) to 34% (range 13-42), P = 0.62, reflecting intestinal absorption. The median ferritin level remained stable 80 μg/L (35-293) vs 82 μg/L (range 37-496) between M0 and M3, P = 0.43. During the 3-month study, median hemoglobin level increased from 11.5 d/dL (range10.4-13.7) to 12.8 g/dL (range 11.1-15.2), P = 0.01. No major side effects were observed. Quality of life assessed by the SF-36 criteria was similar during the 3-month study. Oral iron therapy is effective and safe in EPO-free patients with normal hepcidin levels. These findings suggest that serum hepcidin may be a marker for defining iron therapy strategies in HD patients. HD patients treated with rHU-EPO and with normal hepcidin levels could benefit from oral iron treatment.

Iron metabolism and iron-refractory iron deficiency anemia

Hepcidin is a key molecule that regulates iron metabolism in the body. Iron refractory iron deficiency anemia (IRIDA) is a genetic disorder caused by a defect in the TMPRSS6 gene encoding matriptase-2, a transmembrane serine protease that physiologically inhibits hepcidin production. In patients with IRIDA, the iron uptake in the intestine is remarkably reduced, and iron deficiency anemia (IDA) develops. However, in contrast to the ordinary IDA, high hepcidin levels in IRIDA keep the serum ferritin levels normal or sometimes high. Due to the malabsorption of iron in the intestine, IRIDA is refractory to oral iron supplementation, but partially responds to parenteral iron administration. A high hepcidin level gives IRIDA a lot of similarities with anemia of chronic disease, and a differential diagnosis between the two disorders needs careful inspection. Diagnosis of IRIDA needs genetic testing that is hardly available in most facilities, and therefore its clinical features are not fully understood.

The target hemoglobin content values of reticulocytes for efficient anemia improvement are achieved by low ferritin levels and moderate transferrin saturation: a retrospective observational study

ABSTRACT Objectives: The optimal iron level in hemodialysis (HD) patients remains unclear. The hemoglobin content of reticulocytes (CHr) is a sensitive indicator of iron used for hematopoiesis. To identify the optimal iron content for HD patients, we investigated the relation between CHr levels and iron status, as well as the levels of hepcidin, a main regulator of iron metabolism. Methods: This study enrolled 181 HD outpatients treated with recombinant human erythropoietin (rHuEPO). A sensitivity analysis, using a generalized linear regression model that included the interaction term, was applied to determine the correlations between levels of CHr and those of serum ferritin (s-ft), transferrin saturation (TSAT), and hepcidin. Results: The greatest changes in correlation coefficients for levels of s-ft and TSAT with CHr levels indicated optimal cut-off points of 50 ng/mL (≤50 ng/mL, r = 0.47 vs >50 ng/mL, r = 0.22) and 24% (≤24%, r = 0.58 vs >24%, r = 0.08), respectively. The correlation coefficient for levels of CHr and hepcidin showed that the optimal lower and upper cut-off points were 20 ng/mL (≤20 ng/mL, r = 0.52 vs >20 ng/mL, r = −0.01) and 70 ng/mL (≤70 ng/mL, r = 0.36 vs >70 ng/mL, r = −0.45), respectively. Discussion: This study indicates that the amount of iron in HD patients is sufficient for hematopoiesis under conditions of low s-ft and moderate TSAT levels. High levels of hepcidin could induce negative iron metabolism in hematopoiesis. Conclusion: Therefore, controlling hepcidin levels to within approximately 20–70 ng/mL may prevent iron deficiency and reduced Hb synthesis, and may thus facilitate effective iron utilization in hematopoiesis.