Dimethyl fumarate increases fetal hemoglobin, provides heme detoxification, and corrects anemia in sickle cell disease
Sriram Krishnamoorthy, … , William E. Hobbs, David R. Light
Sriram Krishnamoorthy, … , William E. Hobbs, David R. Light
Published October 19, 2017
Citation Information: JCI Insight. 2017;2(20):e96409. https://doi.org/10.1172/jci.insight.96409.
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Research Article Hematology Inflammation

Dimethyl fumarate increases fetal hemoglobin, provides heme detoxification, and corrects anemia in sickle cell disease

Abstract

Sickle cell disease (SCD) results from a point mutation in the β-globin gene forming hemoglobin S (HbS), which polymerizes in deoxygenated erythrocytes, triggering recurrent painful vaso-occlusive crises and chronic hemolytic anemia. Reactivation of fetal Hb (HbF) expression ameliorates these symptoms of SCD. Nuclear factor (erythroid derived-2)–like 2 (Nrf2) is a transcription factor that triggers cytoprotective and antioxidant pathways to limit oxidative damage and inflammation and increases HbF synthesis in CD34+ stem cell–derived erythroid progenitors. We investigated the ability of dimethyl fumarate (DMF), a small-molecule Nrf2 agonist, to activate γ-globin transcription and enhance HbF in tissue culture and in murine and primate models. DMF recruited Nrf2 to the γ-globin promoters and the locus control region of the β-globin locus in erythroleukemia cells, elevated HbF in SCD donor–derived erythroid progenitors, and reduced hypoxia-induced sickling. Chronic DMF administration in SCD mice induced HbF and increased Nrf2-dependent genes to detoxify heme and limit inflammation. This improved hematological parameters, reduced plasma-free Hb, and attenuated inflammatory markers. Chronic DMF administration to nonanemic primates increased γ-globin mRNA in BM and HbF protein in rbc. DMF represents a potential therapy for SCD to induce HbF and augment vasoprotection and heme detoxification.

Authors

Sriram Krishnamoorthy, Betty Pace, Dipti Gupta, Sarah Sturtevant, Biaoru Li, Levi Makala, Julia Brittain, Nancy Moore, Benjamin F. Vieira, Timothy Thullen, Ivan Stone, Huo Li, William E. Hobbs, David R. Light

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Figure 1

DMF and MMF induce γ-globin mRNA and increase HbF-positive cells (F-cells) in erythroid progenitor cells derived from SCD donor blood PBMC.

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DMF and MMF induce γ-globin mRNA and increase HbF-positive cells (F-cell...
Erythroid progenitor cells were cultured and differentiated from the peripheral blood mononuclear (PBMC) fraction of sickle cell disease (SCD) donor blood (see Supplemental Table 1) and exposed to test compounds as described in Methods with untreated cells used as controls. (A) Levels of γ-globin and GAPDH mRNA were determined between day 13 and 15 by ddPCR analysis and normalized to DMSO-treated control cells (n = 3–4 donors) (B) Levels of γ-globin, β-globin, and GAPDH mRNA were determined on day 11 by real-time PCR analysis, and the ratio of γ/β-globin mRNA was normalized to control cells (n = 4–6 donors). (C) F-cells were quantified by flow cytometry between day 11 and 15, and the % F-cells was normalized to untreated cells (n = 4–7 donors). Results are presented as mean ± SEM. All P values calculated with student t test, 2-tailed (*P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001). One-way ANOVA testing showed significant differences among means (P < 0.05 for all 3 panels). In addition, P values determined by Bonferroni’s multiple comparison test is also indicated († refers to adjusted P < 0.05 when compared with vehicle group). 1SCD donors from Biogen site, 2SCD donors from Augusta University site.