Proteomic Studies for the Investigation of γ-Globin Induction by Decitabine in Human Primary Erythroid Progenitor Cultures

Abstract

Reactivation of γ-globin is considered a promising approach for the treatment of β-thalassemia and sickle cell disease. Therapeutic induction of γ-globin expression, however, is fraught with lack of suitable therapeutic targets. The aim of this study was to investigate the effects that treatment with decitabine has on the proteome of human primary erythroid cells from healthy and thalassemic volunteers, as a means of identifying new potential pharmacological targets. Decitabine is a known γ-globin inducer, which is not, however, safe enough for clinical use. A proteomic approach utilizing isobaric tags for relative and absolute quantitation (iTRAQ) analysis, in combination with high-pH reverse phase peptide fractionation followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), was employed to investigate the effects of decitabine treatment. Bioinformatics analysis making use of the Database for Annotation, Visualization and Integrated Discovery (DAVID) was employed for functional annotation of the 192 differentially expressed proteins identified. The data are available via ProteomeXchange with identifier PXD006889. The proteins fall into various biological pathways, such as the NF-κB signaling pathway, and into many functional categories including regulation of cell proliferation, transcription factor and DNA binding, protein stabilization, chromatin modification and organization, and oxidative stress proteins.

Human cellular model systems of β-thalassemia enable in-depth analysis of disease phenotype

“…Pomalidomide and Decitabine have both been reported to increase γ-globin expression in human primary erythroid cells 17 , 18 . The HBB −/− cells responded accordingly to both drugs, with significant increase in γ-globin levels (by RP-HPLC, Fig.…”

Plasma proteome profiling combined with clinical and genetic features reveals the pathophysiological characteristics of β-thalassemia

“…For example, LGALS3BP is a negative regulator of the NF-κB signaling pathway ( Hong et al., 2019 ). Evidence indicates that NF-κB plays a crucial role in erythropoiesis ( Jeong et al., 2011 ; Zhang et al., 1998 ) while the induction of Hb F by decitabine may be through the suppression of NF-κB activity ( Theodorou et al., 2020 ). Therefore, increased LGALS3BP protein expression may suggest restricted NF-κB activity and increased Hb F production in the erythroid cells.…”

Large-Scale Drug Screen Identifies FDA-Approved Drugs for Repurposing in Sickle-Cell Disease

“…Specifically, HbF blocks HbS from dimerizing, thereby preventing hemoglobin polymer chaining and decreasing the probability of a vaso-occlusion [1,17]. Hydroxyurea, thalidomide, sodium butyrate, and decitabine, a DNMT1 targeting agent, have been previously shown to work through induction of HbF and subsequent protection against VOCs [5,[18][19][20][21][22].…”