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5-Azacytidine acts directly on both erythroid precursors and progenitors to increase production of fetal hemoglobin.

The Journal of Clinical Investigation, 01 Feb 1985, 75(2):547-557
https://doi.org/10.1172/jci111731 PMID: 2579100 PMCID: PMC423530

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Abstract 

The effect of 5-azacytidine on erythroid precursors and progenitors was studied in nine patients with sickle cell anemia or severe thalassemia. Each patient received the drug intravenously for 5 or 7 d. 5-Azacytidine caused a four- to sixfold increase in gamma-messenger RNA concentration in bone marrow cells of eight of the nine patients and decreased the methylation frequency of a specific cytosine residue in the gamma-globin gene promoter in all nine patients. Within 2 d of the start of drug treatment there was a rise in the percentage of reticulocytes containing fetal hemoglobin (HbF; F-reticulocytes) without a significant change in the total number of reticulocytes, which suggested that there was a direct action of 5-azacytidine on erythroid precursors. Late erythroid progenitors (CFU-E), present in bone marrow after 2 d of drug administration, formed colonies containing an increased amount of HbF as compared with control colonies. Moreover, the number of CFU-E derived colonies was not decreased at these early times, which suggested that there was a direct action of 5-azacytidine on erythroid progenitors in the absence of cytotoxicity. Exposure of normal bone marrow cells in tissue culture to 5-azacytidine for 24 h reproduced both of these effects as judged during subsequent colony formation. The combined direct effects of 5-azacytidine on both the erythroid precursor and progenitor compartments resulted in an increase in HbF synthesis that was sustained for 2-3 wk. Toxicity to bone marrow as reflected by cytoreduction was evident after treatment in some patients but was not accompanied by an increase in HbF production. A correlation was found between the effects of 5-azacytidine on bone marrow, as assessed by in vitro measurements, and the hematological response of the individual patients to drug treatment.

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Logo of jcinvestThe Journal of Clinical Investigation
J Clin Invest. 1985 Feb; 75(2): 547–557.
PMCID: PMC423530
PMID: 2579100

5-Azacytidine acts directly on both erythroid precursors and progenitors to increase production of fetal hemoglobin.

R K Humphries, G Dover, N S Young, J G Moore, S Charache, T Ley, and A W Nienhuis
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Abstract

The effect of 5-azacytidine on erythroid precursors and progenitors was studied in nine patients with sickle cell anemia or severe thalassemia. Each patient received the drug intravenously for 5 or 7 d. 5-Azacytidine caused a four- to sixfold increase in gamma-messenger RNA concentration in bone marrow cells of eight of the nine patients and decreased the methylation frequency of a specific cytosine residue in the gamma-globin gene promoter in all nine patients. Within 2 d of the start of drug treatment there was a rise in the percentage of reticulocytes containing fetal hemoglobin (HbF; F-reticulocytes) without a significant change in the total number of reticulocytes, which suggested that there was a direct action of 5-azacytidine on erythroid precursors. Late erythroid progenitors (CFU-E), present in bone marrow after 2 d of drug administration, formed colonies containing an increased amount of HbF as compared with control colonies. Moreover, the number of CFU-E derived colonies was not decreased at these early times, which suggested that there was a direct action of 5-azacytidine on erythroid progenitors in the absence of cytotoxicity. Exposure of normal bone marrow cells in tissue culture to 5-azacytidine for 24 h reproduced both of these effects as judged during subsequent colony formation. The combined direct effects of 5-azacytidine on both the erythroid precursor and progenitor compartments resulted in an increase in HbF synthesis that was sustained for 2-3 wk. Toxicity to bone marrow as reflected by cytoreduction was evident after treatment in some patients but was not accompanied by an increase in HbF production. A correlation was found between the effects of 5-azacytidine on bone marrow, as assessed by in vitro measurements, and the hematological response of the individual patients to drug treatment.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Boyer SH, Belding TK, Margolet L, Noyes AN. Fetal hemoglobin restriction to a few erythrocytes (F cells) in normal human adults. Science. 1975 Apr 25;188(4186):361–363. [Abstract] [Google Scholar]
  • Dover GJ, Boyer SH. Quantitation of hemoglobins within individual red cells: asynchronous biosynthesis of fetal and adult hemoglobin during erythroid maturation in normal subjects. Blood. 1980 Dec;56(6):1082–1091. [Abstract] [Google Scholar]
  • Papayannopoulou T, Kalmantis T, Stamatoyannopoulos G. Cellular regulation of hemoglobin switching: evidence for inverse relationship between fetal hemoglobin synthesis and degree of maturity of human erythroid cells. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6420–6424. [Europe PMC free article] [Abstract] [Google Scholar]
  • Papayannopoulou TH, Brice M, Stamatoyannopoulos G. Stimulation of fetal hemoglobin synthesis in bone marrow cultures from adult individuals. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2033–2037. [Europe PMC free article] [Abstract] [Google Scholar]
  • Papayannopoulou T, Brice M, Stamatoyannopoulos G. Hemoglobin F synthesis in vitro: evidence for control at the level of primitive erythroid stem cells. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2923–2927. [Europe PMC free article] [Abstract] [Google Scholar]
  • Dover GJ, Ogawa M. Cellular mechanisms for increased fetal hemoglobin production in culture. Evidence for continuous commitment to fetal hemoglobin production during burst formation. J Clin Invest. 1980 Nov;66(5):1175–1178. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kidoguchi K, Ogawa M, Karam JD. Hemoglobin biosynthesis in individual erythropoietic bursts in culture. Studies of adult peripheral blood. J Clin Invest. 1979 Apr;63(4):804–806. [Europe PMC free article] [Abstract] [Google Scholar]
  • Dover GJ, Boyer SH, Zinkham WH. Production of erythrocytes that contain fetal hemoglobin in anemia. Transient in vivo changes. J Clin Invest. 1979 Feb;63(2):173–176. [Europe PMC free article] [Abstract] [Google Scholar]
  • Groudine M, Kohwi-Shigematsu T, Gelinas R, Stamatoyannopoulos G, Papayannopoulou T. Human fetal to adult hemoglobin switching: changes in chromatin structure of the beta-globin gene locus. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7551–7555. [Europe PMC free article] [Abstract] [Google Scholar]
  • van der Ploeg LH, Flavell RA. DNA methylation in the human gamma delta beta-globin locus in erythroid and nonerythroid tissues. Cell. 1980 Apr;19(4):947–958. [Abstract] [Google Scholar]
  • DeSimone J, Heller P, Hall L, Zwiers D. 5-Azacytidine stimulates fetal hemoglobin synthesis in anemic baboons. Proc Natl Acad Sci U S A. 1982 Jul;79(14):4428–4431. [Europe PMC free article] [Abstract] [Google Scholar]
  • Ley TJ, DeSimone J, Anagnou NP, Keller GH, Humphries RK, Turner PH, Young NS, Keller P, Nienhuis AW. 5-azacytidine selectively increases gamma-globin synthesis in a patient with beta+ thalassemia. N Engl J Med. 1982 Dec 9;307(24):1469–1475. [Abstract] [Google Scholar]
  • Ley TJ, DeSimone J, Noguchi CT, Turner PH, Schechter AN, Heller P, Nienhuis AW. 5-Azacytidine increases gamma-globin synthesis and reduces the proportion of dense cells in patients with sickle cell anemia. Blood. 1983 Aug;62(2):370–380. [Abstract] [Google Scholar]
  • Charache S, Dover G, Smith K, Talbot CC, Jr, Moyer M, Boyer S. Treatment of sickle cell anemia with 5-azacytidine results in increased fetal hemoglobin production and is associated with nonrandom hypomethylation of DNA around the gamma-delta-beta-globin gene complex. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4842–4846. [Europe PMC free article] [Abstract] [Google Scholar]
  • Torrealba-de Ron AT, Papayannopoulou T, Knapp MS, Fu MF, Knitter G, Stamatoyannopoulos G. Perturbations in the erythroid marrow progenitor cell pools may play a role in the augmentation of HbF by 5-azacytidine. Blood. 1984 Jan;63(1):201–210. [Abstract] [Google Scholar]
  • Letvin NL, Linch DC, Beardsley GP, McIntyre KW, Nathan DG. Augmentation of fetal-hemoglobin production in anemic monkeys by hydroxyurea. N Engl J Med. 1984 Apr 5;310(14):869–873. [Abstract] [Google Scholar]
  • Kantor JA, Turner PH, Nienhuis AW. Beta Thalassemia: mutations which affect processing of the beta-Globin mRNA precursor. Cell. 1980 Aug;21(1):149–157. [Abstract] [Google Scholar]
  • Favaloro J, Treisman R, Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. [Abstract] [Google Scholar]
  • Treisman R, Proudfoot NJ, Shander M, Maniatis T. A single-base change at a splice site in a beta 0-thalassemic gene causes abnormal RNA splicing. Cell. 1982 Jul;29(3):903–911. [Abstract] [Google Scholar]
  • Ley TJ, Anagnou NP, Pepe G, Nienhuis AW. RNA processing errors in patients with beta-thalassemia. Proc Natl Acad Sci U S A. 1982 Aug;79(15):4775–4779. [Europe PMC free article] [Abstract] [Google Scholar]
  • Maxam AM, Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. [Abstract] [Google Scholar]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [Abstract] [Google Scholar]
  • Jeffreys AJ, Flavell RA. The rabbit beta-globin gene contains a large large insert in the coding sequence. Cell. 1977 Dec;12(4):1097–1108. [Abstract] [Google Scholar]
  • Dover GJ, Boyer SH, Bell WR. Microscopic method for assaying F cell production: illustrative changes during infancy and in aplastic anemia. Blood. 1978 Oct;52(4):664–672. [Abstract] [Google Scholar]
  • Gregory CJ, Eaves AC. Human marrow cells capable of erythropoietic differentiation in vitro: definition of three erythroid colony responses. Blood. 1977 Jun;49(6):855–864. [Abstract] [Google Scholar]
  • Finch CA, Deubelbeiss K, Cook JD, Eschbach JW, Harker LA, Funk DD, Marsaglia G, Hillman RS, Slichter S, Adamson JW, et al. Ferrokinetics in man. Medicine (Baltimore) 1970 Jan;49(1):17–53. [Abstract] [Google Scholar]
  • ALPEN EL, CRANMORE D. Cellular kinetics and iron utilization in bone marrow as observed by Fe59 radioautography. Ann N Y Acad Sci. 1959 Jun 25;77:753–765. [Abstract] [Google Scholar]
  • Tanaka K, Appella E, Jay G. Developmental activation of the H-2K gene is correlated with an increase in DNA methylation. Cell. 1983 Dec;35(2 Pt 1):457–465. [Abstract] [Google Scholar]
  • Ley TJ, Chiang YL, Haidaris D, Anagnou NP, Wilson VL, Anderson WF. DNA methylation and regulation of the human beta-globin-like genes in mouse erythroleukemia cells containing human chromosome 11. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6618–6622. [Europe PMC free article] [Abstract] [Google Scholar]
  • Papayannopoulou T, Torrealba de Ron A, Veith R, Knitter G, Stamatoyannopoulos G. Arabinosylcytosine induces fetal hemoglobin in baboons by perturbing erythroid cell differentiation kinetics. Science. 1984 May 11;224(4649):617–619. [Abstract] [Google Scholar]
  • Dover GJ, Boyer SH, Charache S, Heintzelman K. Individual variation in the production and survival of F cells in sickle-cell disease. N Engl J Med. 1978 Dec 28;299(26):1428–1435. [Abstract] [Google Scholar]
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