Induction of erythroferrone in healthy humans by micro-dose recombinant erythropoietin or high-altitude exposure

The erythropoietin (Epo)-erythroferrone (ERFE)-hepcidin axis coordinates erythropoiesis and iron homeostasis. While mouse studies have established that Epo-induced ERFE production represses hepcidin synthesis by inhibiting hepatic BMP/SMAD signaling, evidence for the role of ERFE in humans is limited. To investigate the role of ERFE as a physiological erythroid regulator in humans, we conducted two studies. First, 24 males were given six injections of saline (placebo), recombinant Epo (rhEpo) at a dose of 20 IU/kg (micro-dose) or rhEpo at 50 IU/kg (low dose). Second, we quantified ERFE in 22 subjects exposed to high altitude (3800 m) for 15 h. In the first study, total hemoglobin mass (Hbmass) increased after low- but not after micro-dose injections, when compared to the mass after placebo injections. Serum ERFE levels were enhanced by rhEpo, remaining higher than after placebo for 48 h (micro-dose) or 72 h (low-dose) after injections. Conversely, hepcidin levels decreased when Epo and ERFE rose, before any changes in serum iron parameters occurred. In the second study, serum Epo and ERFE increased at high altitude. The present results demonstrate that in healthy humans ERFE responds to slightly increased Epo levels not associated with Hbmass expansion and downregulates hepcidin in an apparently iron-independent way. Notably, ERFE flags micro-dose Epo, thus holding promise as a novel biomarker of doping.