Type: Oral
Session: 102. Iron Homeostasis and Biology: Exploring Molecular Mechanisms and Therapeutic Options in Iron Homeostasis
Hematology Disease Topics & Pathways:
hematopoiesis, Biological Processes
To understand the mechanisms leading to iron deficiency anemia, as well as changes in the island, we first undertook a comprehensive analysis of hematopoietic progenitors. We measured the relative amounts of megakaryocytic-erythroid progenitors (MEP), granulocyte-monocyte progenitors (GMP) and common myeloid progenitors (CMP) in the bone marrow and observed decreased MEP (WT: 82.81% vs KO: 76.28%, p-value 0.011), increased GMP (WT: 14.94% vs KO: 22.16%, p-value 0.002) and unchanged CMP compared to wild-type (WT) as the mice aged. Given that Irf5 is expressed in both hematopoietic and non-hematopoietic cells, we generated Irf5fl/fl-Vav-iCre+ mice to evaluate the contribution of non-hematopoietic Irf5 to EMBI formation and composition. We observed a significant reduction in the number of EMBI, with overrepresentation of myeloid cells, suggesting that the phenotype observed in Irf5-/- is mostly due to hematopoietic Irf5. However, when we measured the relative amounts of MEP and GMP in the Irf5fl/fl-Vav-iCre+ mice, these remained unchanged, suggesting that cell non-autonomous functions contribute to EMBI composition.
scRNAseq analyses of EMBIs from 3 and 9 months-old littermate-matched WT and Irf5-/- mice revealed that heme synthesis was the only downregulated pathway. In addition, central macrophages from Irf5-/- EMBIs presented decreased expression of major iron regulatory proteins, including heme transporter Slc48a1 and Heme-Oxygenase-1 (HMOX1). Functional analyses using Calcein-AM staining and flow cytometry of freshly isolated central macrophages from enriched EMBIs of WT and Irf5-/- mice revealed that the labile iron pool (LIP) of the central macrophage of Irf5-/- EMBIs was significantly reduced (mean 14.0) compared to WT (mean 49.0, p-value 0.004). This apparent decrease in intracellular iron was concomitantly supported by significant decreases in serum iron (WT: 67.27 uM/L vs KO: 39.67 uM/L, p-value: 0.0059), increased serum EPO (WT: 108 pg/dL vs KO: 163.8 pg/dL, p-value: .0075), and Prussian blue staining of Irf5-/- spleen indicating iron deposition in aging Irf5-/- mice. Finally, these findings were conserved in Irf5fl/fl-Vav-iCre+ mice, highlighting a cell autonomous function for Irf5 in iron regulation within the island.
Together, these data suggest that Irf5 may be a new regulator of iron scavenging, trafficking, and/or utilization within the central macrophage, potentially playing a major role in the red-cell production capacity of the EMBI.
Disclosures: Kalfa: Forma/Novo Nordisk: Consultancy, Research Funding; Agios Pharmaceuticals, Inc.: Consultancy, Research Funding.