{"doi":"10.1111/bjh.18044","title":"Hepcidin is elevated in primary and secondary myelofibrosis and remains elevated in patients treated with ruxolitinib","abstract":"Hepcidin, a peptide hormone produced by hepatocytes, is a key regulator of iron homeostasis in mammals and an important mediator in the innate immune system.1, 2 Pathologically elevated hepcidin levels can cause iron-restricted erythropoiesis and anaemia.3 Myelofibrosis (MF) is a Philadelphia-chromosome-negative myeloproliferative neoplasm (MPN) that is characterized by bone-marrow fibrosis.4 Anaemia is a common problem in MF and is associated with inferior quality-of-life measures and poor prognosis.5, 6 To further assess the relationship between hepcidin and anaemia in MF, we measured serum hepcidin levels in 99 MF patients including 49 primary (PMF) and 50 secondary MF (SMF) patients. Twenty-four of these patients received ruxolitinib therapy, a frequently prescribed JAK1/2 inhibitor for treatment of MF. We also evaluated the relationship between hepcidin and haemoglobin levels, inflammatory cytokines, serum iron markers and erythroferrone. MF patient peripheral-blood (PB) or bone-marrow (BM) samples and healthy donor control PB samples were obtained with written consent according to a protocol approved by the Washington University Human Studies Committee (WU no. 01–1014). Normal control samples were extracted directly from PB. Clinical and genetic information for patients included in the study was obtained through retrospective chart review by the authors Amy Zhou, Maggie J. Allen, and Stephen T. Oh. Statistical analysis was performed using GraphPad Prism (GraphPad Software, La Jolla, CA, USA). The Mann–Whitney U test and Spearman correlations were performed as indicated. Patient characteristics are summarized in Table 1. Hepcidin levels (ng/ml) were measured using a commercially available enzyme-linked immunosorbent assay (ELISA) assay from Intrinsic Lifesciences, La Jolla, CA, USA (Intrinsic Hepcidin IDx™ ELISA) in 99 MF patient plasma samples and nine normal controls. C-reactive protein (CRP), transferrin saturation and ferritin were measured on plasma samples using standard laboratory methods at our institution. Hepcidin was significantly elevated in MF patients compared to normal controls (Figure 1A; median 81.1 vs 8.5, p < 0.0001). There was no difference in hepcidin levels between PMF and SMF patients (Figure 1B; median 103.3 vs 80.8, p = 0.38). A trend between higher hepcidin levels and lower haemoglobin was observed but was not statistically significant (p = 0.719). No correlation was found between hepcidin levels and CRP, transferrin saturation, or ferritin. Hepcidin expression did not differ in samples obtained from patients on treatment with ruxolitinib versus those who were not (median 105.1 vs 80.2, respectively, p = 0.98) (Figure 1C). Erythroferrone (ng/ml) concentrations were quantified using a sandwich ELISA protocol.7 Initially, 96-well plates were coated with a monoclonal capture antibody, washed and blocked. After a 1-h incubation at 25°C with agitation (350 rpm), the plate was washed and incubated for 1 h with 100 μl of biotinylated detection antibody per well (1 μg/ml). The plate was then washed, incubated for 45 min with Neutravidin–horseradish peroxidase conjugate 1/5000 (100 μl per well), and developed with TMB Substrate System for ELISA (ThermoScientific, Waltham, MA, USA) at room temperature for 10 min. The reaction was stopped by adding 50 μl of 2 N sulphuric acid, and the plates were read on a Spectramax 250 (Molecular Devices, San Jose, CA, USA) at 450 nm. Erythroferrone levels were significantly higher in MF patients compared to normal controls (Figure S1A), but no correlation was found between hepcidin and erythroferrone (Figure S1B). When separated by mutational status, no significant difference was observed between hepcidin in JAK2- and CALR-mutated patients, but in non-transfused patients, hepcidin was significantly elevated in CALR- and JAK2-mutated patients compared to triple-negative patients (Figure S1C). Concentrations of 13 cytokines (IFN, IL-10, IL-2, IL-2Rα, IL-6, IL-8, MIP-1α, MI","journal":"British Journal of Haematology","year":2022,"id":254074,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":21,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9642,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":807228,"name":"Tim Kong","orcid":null,"position":1,"is_corresponding":false},{"id":640128,"name":"Jared S. Fowles","orcid":"0000-0002-7614-5447","position":2,"is_corresponding":false},{"id":150727,"name":"Chun‐Ling Jung","orcid":null,"position":3,"is_corresponding":false},{"id":807229,"name":"Maggie J. Allen","orcid":null,"position":4,"is_corresponding":false},{"id":695714,"name":"Daniel A.C. Fisher","orcid":null,"position":5,"is_corresponding":false},{"id":882961,"name":"Mary C. Fulbright","orcid":null,"position":6,"is_corresponding":false},{"id":374946,"name":"Elizabeta Nemeth","orcid":"0000-0002-3477-2397","position":7,"is_corresponding":false},{"id":374947,"name":"Tomas Ganz","orcid":"0000-0002-2830-5469","position":8,"is_corresponding":false},{"id":628540,"name":"Stephen T. Oh","orcid":"0000-0002-8564-5400","position":9,"is_corresponding":false},{"id":900608,"name":"Amy Zhou","orcid":"0000-0003-3259-1995","position":0,"is_corresponding":true}],"reference_count":14,"raw_metadata":null,"created_at":"2026-07-19T00:25:03.918844Z","pmid":"35128632","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}