ECTS2013 Poster Presentations Calciotropic and phosphotropic hormones and mineral metabolism (33 abstracts)
A sensitive assay for measuring circulating BMP6
Martina Pauk 1 , Lovorka Grgurevic 1 , Jelena Brkljacic 1 , Vera Kufner 1 , Tatjana Bordukalo-Niksic 1 , Morana Jankolija 2 , Hermann Oppermann 2 & Slobodan Vukicevic 1
1Laboratory for Mineralized Tissues, School of Medicine, Center for Translational and Clinical Research, University of Zagreb, Zagreb, Croatia; 2Genera Research, Rakov Potok, Croatia.
Although bone morphogenetic protein 6 (BMP6) is known for its ability to induce growth of bone and cartilage, recent studies identified BMP6 as a key endogenous regulator of hepcidin expression and iron metabolism. Here, we examined BMP6 presence in the serum and investigated whether circulating levels of BMP6 may reflect body iron status. First, we showed by liquid chromatography–mass spectrometry (LC–MS) and western blotting that BMP6 is present in the circulation of healthy humans. We next analyzed biological fluids of mouse and human using commercial ELISAs, but failed to detect BMP6. To enhance the assay sensitivity and simplify the BMP6 measurement procedure, we developed a BMP6 Proximity Extension Assay (PEA) which enabled us to analyze up to 10 pg/ml of BMP6 in serum samples before and following iron exposure. We determined that BMP6 circulates at 55.46±9.8 pg/ml, and this is the first demonstration of a physiological range of circulating BMP6 in mice. Interestingly, in untreated mice BMP6 concentrations displayed a diurnal variation, with concentrations being lowest in the early morning and increasing throughout the day before declining during the evening hours. Given the already known hepcidin diurnal rhythm, our results indicate that hepcidin variations are in fact reflecting BMP6 pattern. Furthermore, we found that iron loading was followed by a BMP6 increase in mouse serum at 12 h to 109.96±25.4 and at 24 h to 128.7±21 pg/ml, indicating that circulating BMP6 is likely to play a role in iron metabolism. As circulating BMP6 positively correlates with iron, the measurement of BMP6 in biological fluids presents a promising tool in the diagnosis of conditions in which iron metabolism is affected. Therefore, the development and validation of BMP6 assays would increase our understanding of the physiology of iron homeostasis and iron related bone loss.
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Bone Abstracts
ISSN 2052-1219 (online)
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