Thalassemias are a heterogeneous group of red blood cell disorders, considered a major cause of morbidity and mortality among genetic diseases. However, there is still no universally available cure for thalassemias. The underlying basis of thalassemia pathology is the premature apoptotic destruction of erythroblasts causing ineffective erythropoiesis. In β-thalassemia, β-globin synthesis is reduced causing α-globin accumulation. Unpaired globin chains, with heme attached to them, accumulate in thalassemic erythroblasts causing oxidative stress and the premature cell death. We hypothesize that in β-thalassemia heme oxygenase (HO) 1 could play a pathogenic role in the development of anemia and ineffective erythropoiesis. To test this hypothesis, we exploited a mouse model of β-thalassemia intermedia, Th3/+ We observed that HO inhibition using tin protoporphyrin IX (SnPP) decreased heme-iron recycling in the liver and ameliorated anemia in the Th3/+ mice. SnPP administration led to a decrease in erythropoietin and increase in hepcidin serum levels, changes that were accompanied by an alleviation of ineffective erythropoiesis in Th3/+ mice. Additionally, the bone marrow from Th3/+ mice treated with SnPP exhibited decreased heme catabolism and diminished iron release as well as reduced apoptosis. Our results indicate that the iron released from heme because of HO activity contributes to the pathophysiology of thalassemia. Therefore, new therapies that suppress heme catabolism may be beneficial in ameliorating the anemia and ineffective erythropoiesis in thalassemias.

Department of Biology Faculty of Medicine and Dentistry Palacky University Olomouc Czech Republic

Department of Physiology McGill University Montreal QC Canada; and

Lady Davis Institute for Medical Research Jewish General Hospital Montreal QC Canada

Zobrazit více v PubMed

Musallam KM, Rivella S, Vichinsky E, Rachmilewitz EA. Non-transfusion-dependent thalassemias. Haematologica. 2013;98(6):833-844. PubMed PMC

Yang B, Kirby S, Lewis J, Detloff PJ, Maeda N, Smithies O. A mouse model for beta 0-thalassemia. Proc Natl Acad Sci USA. 1995;92(25):11608-11612. PubMed PMC

Drakesmith H, Nemeth E, Ganz T. Ironing out ferroportin. Cell Metab. 2015;22(5):777-787. PubMed PMC

Maines MD. The heme oxygenase system: a regulator of second messenger gases. Annu Rev Pharmacol Toxicol. 1997;37(1):517-554. PubMed

Maines MD. The heme oxygenase system: update 2005. Antioxid Redox Signal. 2005;7(11-12):1761-1766. PubMed

Poss KD, Tonegawa S. Heme oxygenase 1 is required for mammalian iron reutilization. Proc Natl Acad Sci USA. 1997;94(20):10919-10924. PubMed PMC

Abraham NG, Kappas A. Pharmacological and clinical aspects of heme oxygenase. Pharmacol Rev. 2008;60(1):79-127. PubMed

Shibahara S, Müller RM, Taguchi H. Transcriptional control of rat heme oxygenase by heat shock. J Biol Chem. 1987;262(27):12889-12892. PubMed

Keyse SM, Tyrrell RM. Heme oxygenase is the major 32-kDa stress protein induced in human skin fibroblasts by UVA radiation, hydrogen peroxide, and sodium arsenite. Proc Natl Acad Sci USA. 1989;86(1):99-103. PubMed PMC

Nascimento AL, Luscher P, Tyrrell RM. Ultraviolet A (320-380 nm) radiation causes an alteration in the binding of a specific protein/protein complex to a short region of the promoter of the human heme oxygenase 1 gene. Nucleic Acids Res. 1993;21(5):1103-1109. PubMed PMC

Garcia-Santos D, Schranzhofer M, Horvathova M, et al. . Heme oxygenase 1 is expressed in murine erythroid cells where it controls the level of regulatory heme. Blood. 2014;123(14):2269-2277. PubMed

Neuwirt J, Ponka P, Borová J. Evidence for the presence of free and protein-bound nonhemoglobin heme in rabbit reticulocytes. Biochim Biophys Acta. 1972;264(2):235-244. PubMed

Garrick MD, Scott D, Kulju D, Romano MA, Dolan KG, Garrick LM. Evidence for and consequences of chronic heme deficiency in Belgrade rat reticulocytes. Biochim Biophys Acta. 1999;1449(2):125-136. PubMed

Lok CN, Ponka P. Identification of an erythroid active element in the transferrin receptor gene. J Biol Chem. 2000;275(31):24185-24190. PubMed

Tahara T, Sun J, Igarashi K, Taketani S. Heme-dependent up-regulation of the alpha-globin gene expression by transcriptional repressor Bach1 in erythroid cells. Biochem Biophys Res Commun. 2004;324(1):77-85. PubMed

Tahara T, Sun J, Nakanishi K, et al. . Heme positively regulates the expression of beta-globin at the locus control region via the transcriptional factor Bach1 in erythroid cells. J Biol Chem. 2004;279(7):5480-5487. PubMed

Chen JJ. Regulation of protein synthesis by the heme-regulated eIF2alpha kinase: relevance to anemias. Blood. 2007;109(7):2693-2699. PubMed PMC

Ponka P. Tissue-specific regulation of iron metabolism and heme synthesis: distinct control mechanisms in erythroid cells. Blood. 1997;89(1):1-25. PubMed

Schrier SL. Pathophysiology of thalassemia. Curr Opin Hematol. 2002;9(2):123-126. PubMed

Liu B, Qian JM. Cytoprotective role of heme oxygenase-1 in liver ischemia reperfusion injury. Int J Clin Exp Med. 2015;8(11):19867-19873. PubMed PMC

Chang M, Xue J, Sharma V, Habtezion A. Protective role of hemeoxygenase-1 in gastrointestinal diseases. Cell Mol Life Sci. 2015;72(6):1161-1173. PubMed PMC

Son Y, Lee JH, Cheong YK, Chung HT, Pae HO. Antidiabetic potential of the heme oxygenase-1 inducer curcumin analogues. Biomed Res Int 2013;2013:918039. PubMed PMC

Wang CF, Wang ZY, Li JY. Dual protective role of HO-1 in transplanted liver grafts: a review of experimental and clinical studies. World J Gastroenterol. 2011;17(26):3101-3108. PubMed PMC

Sassa S. Why heme needs to be degraded to iron, biliverdin IXalpha, and carbon monoxide? Antioxid Redox Signal. 2004;6(5):819-824. PubMed

Stocker R, Yamamoto Y, McDonagh AF, Glazer AN, Ames BN. Bilirubin is an antioxidant of possible physiological importance. Science. 1987;235(4792):1043-1046. PubMed

Rivella S. β-thalassemias: paradigmatic diseases for scientific discoveries and development of innovative therapies. Haematologica. 2015;100(4):418-430. PubMed PMC

Belcher JD, Mahaseth H, Welch TE, Otterbein LE, Hebbel RP, Vercellotti GM. Heme oxygenase-1 is a modulator of inflammation and vaso-occlusion in transgenic sickle mice. J Clin Invest. 2006;116(3):808-816. PubMed PMC

Berglund L, Angelin B, Blomstrand R, Drummond G, Kappas A. Sn-protoporphyrin lowers serum bilirubin levels, decreases biliary bilirubin output, enhances biliary heme excretion and potently inhibits hepatic heme oxygenase activity in normal human subjects. Hepatology. 1988;8(3):625-631. PubMed

Kappas A, Drummond GS, Galbraith RA. Prolonged clinical use of a heme oxygenase inhibitor: hematological evidence for an inducible but reversible iron-deficiency state. Pediatrics. 1993;91(3):537-539. PubMed

Magnani M, Rossi L, Stocchi V, Cucchiarini L, Piacentini G, Fornaini G. Effect of age on some properties of mice erythrocytes. Mech Ageing Dev. 1988;42(1):37-47. PubMed

Sassa S. Sequential induction of heme pathway enzymes during erythroid differentiation of mouse Friend leukemia virus-infected cells. J Exp Med. 1976;143(2):305-315. PubMed PMC

Socolovsky M, Nam H, Fleming MD, Haase VH, Brugnara C, Lodish HF. Ineffective erythropoiesis in Stat5a(-/-)5b(-/-) mice due to decreased survival of early erythroblasts. Blood. 2001;98(12):3261-3273. PubMed

Sorensen S, Rubin E, Polster H, Mohandas N, Schrier S. The role of membrane skeletal-associated alpha-globin in the pathophysiology of beta-thalassemia. Blood. 1990;75(6):1333-1336. PubMed

Kong Y, Zhou S, Kihm AJ, et al. . Loss of alpha-hemoglobin-stabilizing protein impairs erythropoiesis and exacerbates beta-thalassemia. J Clin Invest. 2004;114(10):1457-1466. PubMed PMC

Akel A, Wagner CA, Kovacikova J, et al. . Enhanced suicidal death of erythrocytes from gene-targeted mice lacking the Cl-/HCO(3)(-) exchanger AE1. Am J Physiol Cell Physiol. 2007;292(5):C1759-C1767. PubMed

Soe-Lin S, Apte SS, Andriopoulos B Jr, et al. . Nramp1 promotes efficient macrophage recycling of iron following erythrophagocytosis in vivo. Proc Natl Acad Sci USA. 2009;106(14):5960-5965. PubMed PMC

Kina T, Ikuta K, Takayama E, et al. . The monoclonal antibody TER-119 recognizes a molecule associated with glycophorin A and specifically marks the late stages of murine erythroid lineage. Br J Haematol. 2000;109(2):280-287. PubMed

Paulson RF, Shi L, Wu DC. Stress erythropoiesis: new signals and new stress progenitor cells. Curr Opin Hematol. 2011;18(3):139-145. PubMed PMC

Kappas A, Simionatto CS, Drummond GS, Sassa S, Anderson KE. The liver excretes large amounts of heme into bile when heme oxygenase is inhibited competitively by Sn-protoporphyrin. Proc Natl Acad Sci USA. 1985;82(3):896-900. PubMed PMC

Wong RJ, Vreman HJ, Schulz S, Kalish FS, Pierce NW, Stevenson DK. In vitro inhibition of heme oxygenase isoenzymes by metalloporphyrins. J Perinatol. 2011;31(Suppl 1):S35-S41. PubMed

Rivella S. Ineffective erythropoiesis and thalassemias. Curr Opin Hematol. 2009;16(3):187-194. PubMed PMC

Greer JP, Arber DA. Wintrobe’s Clinical Hematology. Philadelphia, PA: Lippincott Williams & Wilkins; 2014.

Kautz L, Jung G, Valore EV, Rivella S, Nemeth E, Ganz T. Identification of erythroferrone as an erythroid regulator of iron metabolism. Nat Genet. 2014;46(7):678-684. PubMed PMC

Kautz L, Jung G, Du X, et al. . Erythroferrone contributes to hepcidin suppression and iron overload in a mouse model of β-thalassemia. Blood. 2015;126(17):2031-2037. PubMed PMC

Weatherall DJ, Clegg JB. The Thalassaemia Syndromes. Hoboken, NJ: Wiley; 2008.

Eisenstein RS, Garcia-Mayol D, Pettingell W, Munro HN. Regulation of ferritin and heme oxygenase synthesis in rat fibroblasts by different forms of iron. Proc Natl Acad Sci USA. 1991;88(3):688-692. PubMed PMC

Casu C, Rivella S. Iron age: novel targets for iron overload. Hematology (Am Soc Hematol Educ Program). 2014;2014(1):216-221. PubMed PMC

Casu C, Oikonomidou PR, Chen H, et al. . Minihepcidin peptides as disease modifiers in mice affected by β-thalassemia and polycythemia vera. Blood. 2016;128(2):265-276. PubMed PMC

Guo S, Casu C, Gardenghi S, et al. . Reducing TMPRSS6 ameliorates hemochromatosis and β-thalassemia in mice. J Clin Invest. 2013;123(4):1531-1541. PubMed PMC

Nai A, Pagani A, Mandelli G, et al. . Deletion of TMPRSS6 attenuates the phenotype in a mouse model of β-thalassemia. Blood. 2012;119(21):5021-5029. PubMed PMC

Schmidt PJ, Racie T, Westerman M, Fitzgerald K, Butler JS, Fleming MD. Combination therapy with a Tmprss6 RNAi-therapeutic and the oral iron chelator deferiprone additively diminishes secondary iron overload in a mouse model of β-thalassemia intermedia. Am J Hematol. 2015;90(4):310-313. PubMed PMC

Schmidt PJ, Toudjarska I, Sendamarai AK, et al. . An RNAi therapeutic targeting Tmprss6 decreases iron overload in Hfe(-/-) mice and ameliorates anemia and iron overload in murine β-thalassemia intermedia. Blood. 2013;121(7):1200-1208. PubMed PMC

Ganz T. Systemic iron homeostasis. Physiol Rev. 2013;93(4):1721-1741. PubMed

Preza GC, Ruchala P, Pinon R, et al. . Minihepcidins are rationally designed small peptides that mimic hepcidin activity in mice and may be useful for the treatment of iron overload. J Clin Invest. 2011;121(12):4880-4888. PubMed PMC

Delaby C, Pilard N, Puy H, Canonne-Hergaux F. Sequential regulation of ferroportin expression after erythrophagocytosis in murine macrophages: early mRNA induction by haem, followed by iron-dependent protein expression. Biochem J. 2008;411(1):123-131. PubMed

Finch CA, Sturgeon P. Erythrokinetics in Cooley’s anemia. Blood. 1957;12(1):64-73. PubMed

Pootrakul P, Huebers HA, Finch CA, Pippard MJ, Cazzola M. Iron metabolism in thalassemia. Birth Defects Orig Artic Ser. 1988;23(5B):3-8. PubMed

Fang J, Menon M, Kapelle W, et al. . EPO modulation of cell-cycle regulatory genes, and cell division, in primary bone marrow erythroblasts. Blood. 2007;110(7):2361-2370. PubMed PMC

Socolovsky M, Murrell M, Liu Y, Pop R, Porpiglia E, Levchenko A. Negative autoregulation by FAS mediates robust fetal erythropoiesis. PLoS Biol. 2007;5(10):e252. PubMed PMC

Menon MP, Karur V, Bogacheva O, Bogachev O, Cuetara B, Wojchowski DM. Signals for stress erythropoiesis are integrated via an erythropoietin receptor-phosphotyrosine-343-Stat5 axis. J Clin Invest. 2006;116(3):683-694. PubMed PMC

Theurl I, Hilgendorf I, Nairz M, et al. . On-demand erythrocyte disposal and iron recycling requires transient macrophages in the liver. Nat Med. 2016;22(8):945-951. PubMed PMC

Ponka P, Sheftel AD, English AM, Scott Bohle D, Garcia-Santos D. Do mammalian cells really need to export and import heme? Trends Biochem Sci. 2017;42(5):395-406. PubMed

Schulz S, Wong RJ, Vreman HJ, Stevenson DK. Metalloporphyrins - an update. Front Pharmacol. 2012;3:68. PubMed PMC

Anderson KE, Simionatto CS, Drummond GS, Kappas A. Disposition of tin-protoporphyrin and suppression of hyperbilirubinemia in humans. Clin Pharmacol Ther. 1986;39(5):510-520. PubMed