Trienone analogs of curcuminoids induce fetal hemoglobin synthesis via demethylation at Gγ-globin gene promoter

Abstract

The reactivation of γ-globin chain synthesis to combine with excess free α-globin chains and form fetal hemoglobin (HbF) is an important alternative treatment for β-thalassemia. We had reported HbF induction property of natural curcuminoids, curcumin (Cur), demethoxycurcumin (DMC) and bis-demethoxycurcumin (BDMC), in erythroid progenitors. Herein, the HbF induction property of trienone analogs of the three curcuminoids in erythroleukemic K562 cell lines and primary human erythroid progenitor cells from β-thalassemia/HbE patients was examined. All three trienone analogs could induce HbF synthesis. The most potent HbF inducer in K562 cells was trienone analog of BDMC (T-BDMC) with 2.4 ± 0.2 fold increase. In addition, DNA methylation at CpG − 53, − 50 and + 6 of Gγ-globin gene promoter in K562 cells treated with the compounds including T-BDMC (9.3 ± 1.7%, 7.3 ± 1.7% and 5.3 ± 0.5%, respectively) was significantly lower than those obtained from the control cells (30.7 ± 3.8%, 25.0 ± 2.9% and 7.7 ± 0.9%, respectively P < 0.05). The trienone compounds also significantly induced HbF synthesis in β-thalassemia/HbE erythroid progenitor cells with significantly reduction in DNA methylation at CpG + 6 of Gγ-globin gene promoter. These results suggested that the curcuminoids and their three trienone analogs induced HbF synthesis by decreased DNA methylation at Gγ-globin promoter region, without effect on Aγ-globin promoter region.

The Long Scientific Journey of Sirolimus (Rapamycin): From the Soil of Easter Island (Rapa Nui) to Applied Research and Clinical Trials on β-Thalassemia and Other Hemoglobinopathies

“…Key studies that demonstrated the usefulness of K562 cells for the screening of HbF inducers were published by Rutherford et al [ 82 ], who investigated the capacity of these cells to differentiate following treatment with hemin, producing large amounts of hemoglobin, mainly the embryo-fetal hemoglobins Hb Gower 1 (α 2 ε 2 ), Hb Portland (ζ 2 γ 2 ), and to a lesser extent HbF (α 2 γ 2 ). Accordingly, the erythroleukemia K562 cell line was used in several studies for the first screening of potential inducers of the expression of γ-globin genes and accumulation of fetal hemoglobin [ 83 , 84 , 85 , 86 , 87 , 88 ].…”

Expression of γ-globin genes in β-thalassemia patients treated with sirolimus: results from a pilot clinical trial (Sirthalaclin)

“…[13][14][15][16][17] Accordingly, these observations have prompted research on inducers of HbF that can therapeutically reproduce what occurs in β-thalassemia patients, with natural persistence of higher levels of HbF. [18][19][20][21] Furthermore, genome editing approaches are available for the induction of HbF following the elimination of genomic sequences encoding for transcriptional repressors or genomic sequences targeted by these regulatory factors. [22][23][24][25] In this context, clinical trials are underway, such as NCT03655678 (A safety and efficacy study evaluating CTX001 in subjects with TDT), based on the use of autologous CRISPR-Cas9modified CD34 + human hematopoietic stem and progenitor cells with CTX001.…”

Combined approaches for increasing fetal hemoglobin (HbF) and de novo production of adult hemoglobin (HbA) in erythroid cells from β-thalassemia patients: treatment with HbF inducers and CRISPR-Cas9 based genome editing

“…More recent clinical studies support the concept that naturally higher production of HbF improves the clinical phenotype of a variety of β-thalassemia patients ( Uda et al, 2008 ; Galanello et al, 2009 ; Nuinoon et al, 2010 ; Badens et al, 2011 ; Danjou et al, 2012 ). Accordingly, these observations have prompted the activity of a large number of research groups in performing studies on inducers of HbF that can be proposed in clinical trials, as they reproduce to some extent what occurs in β-thalassemia patients with a natural persistence of higher levels of HbF ( Gambari and Fibach, 2007 ; Finotti and Gambari, 2014 ; Lavelle et al, 2018 ; Nuamsee et al, 2021 ). In addition to HbF inducers, reactivation of HbF production can be obtained using different CRISPR-Cas9-based gene editing approaches ( Figure 2 ).…”