Curcumin as an Epigenetic Therapeutic Agent in Myelodysplastic Syndromes (MDS)

Abstract

Growth Factor Independence 1 (GFI1) is a transcription factor with an important role in the regulation of development of myeloid and lymphoid cell lineages and was implicated in the development of myelodysplastic syndrome (MDS) and acute myeloid leukaemia (AML). Reduced expression of GFI1 or presence of the GFI1-36N (serine replaced with asparagine) variant leads to epigenetic changes in human and murine AML blasts and accelerated the development of leukaemia in a murine model of human MDS and AML. We and other groups previously showed that the GFI1-36N allele or reduced expression of GFI1 in human AML blasts is associated with an inferior prognosis. Using GFI1-36S, -36N -KD, NUP98-HOXD13-tg mice and curcumin (a natural histone acetyltransferase inhibitor (HATi)), we now demonstrate that expansion of GFI1-36N or -KD, NUP98-HODXD13 leukaemic cells can be delayed. Curcumin treatment significantly reduced AML progression in GFI1-36N or -KD mice and prolonged AML-free survival. Of note, curcumin treatment had no effect in GFI1-36S, NUP98-HODXD13 expressing mice. On a molecular level, curcumin treatment negatively affected open chromatin structure in the GFI1-36N or -KD haematopoietic cells but not GFI1-36S cells. Taken together, our study thus identified a therapeutic role for curcumin treatment in the treatment of AML patients (homo or heterozygous for GFI1-36N or reduced GFI1 expression) and possibly improved therapy outcome.

Keywords: GFI1; acute myeloid leukaemia; curcumin; histone acetyltransferase inhibitor.

Figure A1

Overview of overall survival GFI1-36S, -36N and –KD, NUP98-HOXD13-tg mice.

Figure A2

Curcumin treatment improved the overall survival of -36N and –KD, NUP98-HOXD13-tg mice. AML free survival in curcumin-treated GFI1-36S and -36N, NUP98-HOXD13-tg mice (A) or GFI1-36S and -KD, NUP98-HOXD13-tg mice (B) are presented.

Figure A3

Curcumin treatment reduced leukemic blasts in GFI1-36N and -KD mice. (A) Frequency of leukemic blasts (Wright-Giemsa staining) in BM cytospins derived from GFI1-36S/-36N/-KD, NUP98/HOXD13-tg leukaemic mice (n = 4–7 per group). (B) Representative images in different groups. Scale bars represent 20 µm respectively.

Figure A4

GSEA analysis based on ChIP-seq data identified differential regulation of hallmark heme-metabolism in curcumin-treated GFI1-36N and -KD, NUP98-HOXD13-tg mice. Representative hallmark heme-metabolism enrichment plots in GFI1-36N (A), -KD (B) and -36S (C), NUP98-HOXD13-tg mice.

Figure 1

Effect of curcumin on AML free survival of GFI1-36S/ -36N/-KD, NUP98/HOXD13-tg mice. (A–C). Overview of the AML-free survival in GFI1-36S (A), GFI1-36N (B) and GFI1-KD (C) mice treated with curcumin or vehicle. (D). Quantification of AML development in vehicle or curcumin-treated GFI1-36S, -36N and -KD, GFI1-36S/ -36N/-KD, NUP98/HOXD13-tg mice. * p < 0.05.

Figure 2

Curcumin treatment affected parameters contributing to AML development. (A,B). Percentage of c-kit positive (c-kit+) cells in the bone marrow (BM) and spleen in the vehicle and curcumin-treated GFI1-36S/ -36N and -KD, NUP98/HOXD13-tg mice. (C,D). Quantification of platelets and haemoglobin in curcumin and vehicle-treated GFI1-36S/ -36N and -KD, NUP98/HOXD13-tg mice. **** p < 0.001, *** p < 0.005, ** p < 0.01. ns: not significant.

Figure 3

Curcumin treatment affected GFI1 promoter occupancy in the c-kit+ cells GFI1-36S/-36N and -KD, NUP98/HOXD13-tg mice. CHIP sequencing was performed in the frozen BM cells of GFI1-36S/ -36N and -KD, NUP98/HOXD13-tg mice. The mean and log2 fold change of peak counts (promoter ±1 kb) in GFI1-36S (A), -36N (B) and –KD (C), NUP98/HOXD13-tg mice (vehicle vs curcumin treatment) are shown. Peak counts were calculated as normalized read counts for each consensus peak located in promoter regions (±1 kb). Individual points are coloured by the number of neighbouring points to display the point density.

Figure 4

Curcumin treatment significantly affected hallmark Heme-metabolism in GFI1-36N/-KD, NUP98-HOXD13-tg mice. Significantly enriched (FDR q-value < 0.05) hallmark gene sets between curcumin-treated and untreated samples are presented. Genes were ranked by log 2 fold change of normalized peak read counts between curcumin-treated and untreated samples. For genes with multiple associated peaks, the peak with the largest absolute fold change was used. The colour of the dot denotes the FDR q-value and the size of the absolute value of the normalized enrichment score. The columns from left to right respectively demonstrate negatively enriched gene sets in GFI1-36N/ -KD and positively enriched gene sets in GFI1-36S, NUP98-HOXD13-tg curcumin-treated samples.