Alternative options for DNA-based experimental therapy of β-thalassemia

Expert Opinion on Biological Therapy

Volumn 12, Issue 4, 2012, Pages 443-462

  Gambari, Roberto ^a  

^a UNIVERSITY OF FERRARA   (Italy)

Author keywords

Antisense; Gene therapy; HbF induction; Induced pluripotent cells; MicroRNA; Thalassemia; Transcription factor decoy; Transcription factors

Indexed keywords

5 AZA 2' DEOXYCYTIDINE; ALPHA GLOBIN; AZACITIDINE; BETA GLOBIN; CHROMOMYCIN; CISPLATIN; CYTARABINE; GAMMA GLOBIN; HEMOGLOBIN F; HYDROXYUREA; LENTIVIRUS VECTOR; MICRORNA; MITHRAMYCIN; RETROVIRUS VECTOR; TALLIMUSTINE; THALIDOMIDE; TRANSCRIPTION FACTOR;

BETA THALASSEMIA; BLOOD TRANSFUSION; CELL THERAPY; CHELATION THERAPY; DNA MODIFICATION; EMBRYONIC STEM CELL; ERYTHROID CELL; ERYTHROID PRECURSOR CELL; EXPERIMENTAL THERAPY; GENE EXPRESSION; GENE INSERTION; GENE MUTATION; GENE TARGETING; GENE THERAPY; GENETIC TRANSCRIPTION; GENOTOXICITY; HUMAN; IN VITRO STUDY; IN VIVO STUDY; MOLONEY LEUKEMIA ONCOVIRUS; NONHUMAN; NUCLEAR REPROGRAMMING; PHENOTYPE; PLURIPOTENT STEM CELL; REVIEW; RNA REPAIR; SOMATIC CELL; TARGET CELL; TARGETED GENE REPAIR;

ADULT; BETA-THALASSEMIA; GENE THERAPY; GENETIC VECTORS; HUMANS;

EID: 84858643077     PISSN: 14712598     EISSN: None     Source Type: Journal    
DOI: 10.1517/14712598.2012.665047     Document Type: Review

References (148)

1. Patrinos GP, Kollia P, Papadakis MN. Molecular diagnosis of inherited disorders: lessons from hemoglobinopathies. Hum Mutat 2005;26:399-412 (Pubitemid 41572447)
2. Old JM. Screening and genetic diagnosis of haemoglobin disorders. Blood Rev 2003;17:43-53 (Pubitemid 36176210)
3. Galanello R, Origa R. Beta-thalassemia. Orphanet J Rare Dis 2010;5:11
4. Colah R, Gorakshakar A, Nadkarni A. Global burden, distribution and prevention of beta-thalassemias and hemoglobin E disorders. Expert Rev Hematol 2010;3:103-17
5. Quek L, Thein SL. Molecular therapies in beta-thalassaemia. Br J Haematol 2007;136:353-65
6. Lederer CW, Basak AN, Aydinok Y, et al. An electronic infrastructure for research and treatment of the thalassemias and other hemoglobinopathies: the Euro-mediterranean ITHANET project. Hemoglobin 2009;33:163-76
7. Gambari R, Fibach E. Medicinal chemistry of fetal hemoglobin inducers for treatment of beta-thalassemia. Curr Med Chem 2007;14:199-212 (Pubitemid 46043915)
8. Testa U. Fetal hemoglobin chemical inducers for treatment of hemoglobinopathies. Ann Hematol 2009;88:505-28
9. Cousens NE, Gaff CL, Metcalfe SA, Delatycki MB. Carrier screening for beta-thalassaemia: a review of international practice. Eur J Hum Genet 2010;18:1077-83
10. Mantikou E, Arkesteijn SG, Beckhoven Van JM, Kerkhoffs JL. A brief review on newborn screening methods for hemoglobinopathies and preliminary results selecting beta thalassemia carriers at birth by quantitative estimation of the HbA fraction. Clin Biochem 2009;42:1780-5
11. Cao A, Galanello R. Beta-thalassemia. Genet Med 2010;12:61-76
12. Cao A, Moi P, Galanello R. Recent advances in beta-thalassemias. Pediatr Rep 2011;3:e17
13. Gambari R. Foetal haemoglobin inducers and thalassaemia: novel achievements. Blood Transfus 2010;8:5-7
14. Cunningham MJ. Update on thalassemia: clinical care and complications. Hematol Oncol Clin North Am 2010;24:215-27
15. Toumba M, Sergis A, Kanaris C, Skordis N. Endocrine complications in patients with thalassaemia Major. Pediatr Endocrinol Rev 2007;5:642-8
16. Tuck SM. Fertility and pregnancy in thalassemia major. Ann N Y Acad Sci 2005;1054:300-7 (Pubitemid 43031037)
17. Skordis N, Kyriakou A. The multifactorial origin of growth failure in thalassaemia. Pediatr Endocrinol Rev 2011;8(Suppl 2):271-7
18. De Sanctis V. Growth and puberty and its management in thalassaemia. Horm Res 2002;58(Suppl 1):72-9 (Pubitemid 35316841)
19. Borgna-Pignatti C, Gamberini MR. Complications of thalassemia major and their treatment. Expert Rev Hematol 2011;4:353-66
20. De Sanctis V, Gamberini MR, Borgatti L, et al. Alpha and beta cell evaluation in patients with thalassaemia intermedia and iron overload. Postgrad Med J 1985;61:963-7 (Pubitemid 16235249)
21. Di Marco V, Bronte F, Cabibi D, et al. Noninvasive assessment of liver fibrosis in thalassaemia major patients by transient elastography (TE) - lack of interference by iron deposition. Br J Haematol 2010;148:476-9
22. NIH-NHLBI/NIDDK Thalassemia Workshop: clinical Priorities and Clinical Trials, Fishers Lane Conference Center, 5635 Fishers Lane, Rockville, Maryland, May 20 - 21, 2009. Available from: http://www.nhlbi.nih.gov/meetings/ workshops/thal-workshop.htm [Last accessed 13 February 2012]
23. Bianchi N, Zuccato C, Lampronti I, et al. Fetal hemoglobin inducers from the natural world; a novel approach for identification of drugs for the treatment of beta-thalassemia and sickle-cell anemia. Evid Based Complement Altern Med 2009;6:141-51
24. Feriotto G, Salvatori F, Finotti A, et al. A novel frameshift mutation (+A) at codon 18 of the beta-globin gene associated with high persistence of fetal hemoglobin phenotype and deltabeta-thalassemia. Acta Haematol 2008;119:28-37 (Pubitemid 351363855)
25. Zhao H, Pestina TI, Nasimuzzaman M, et al. Amelioration of murine beta-thalassemia through drug selection of hematopoietic stem cells transduced with a lentiviral vector encoding both gamma-globin and the MGMT drug-resistance gene. Blood 2009;113:5747-56
26. Bianchi N, Borgatti M, Fibach E, et al. Bis-epoxyethyl derivatives of distamycin A modified on the amidino moiety: induction of production of fetal hemoglobin in human erythroid precursor cells. Int J Mol Med 2009;23:105-11
27. Guerrini A, Lampronti I, Bianchi N, et al. Bergamot (Citrus bergamia Risso) fruit extracts as gamma-globin gene expression inducers: phytochemical and functional perspectives. J Agric Food Chem 2009;57:4103-11
28. Lampronti I, Bianchi N, Zuccato C, et al. Increase in gamma-globin mRNA content in human erythroid cells treated with angelicin analogs. Int J Hematol 2009;90:318-27
29. Moutouh-de Parseval LA, Verhelle D, Glezer E, et al. Pomalidomide and lenalidomide regulate erythropoiesis and fetal hemoglobin production in human CD34+ cells. J Clin Invest 2008;118:248-58
30. Macari ER, Lowrey CH. Induction of human fetal hemoglobin via the NRF2 antioxidant response signaling pathway. Blood 2011;117:5987-97
31. Boosalis MS, Castaneda SA, Trudel M, et al. Novel therapeutic candidates, identified by molecular modeling, induce gamma-globin gene expression in vivo. Blood Cells Mol Dis 2011;47:107-16
32. Italia KY, Jijina FJ, Merchant R, et al. Response to hydroxyurea in beta thalassemia major and intermedia: experience in western India. Clin Chim Acta 2009;407:10-15
33. Ehsani MA, Hedayati-Asl AA, Bagheri A, et al. Hydroxyurea-induced hematological response in transfusion-independent betathalassemia intermedia: case series and review of literature. Pediatr Hematol Oncol 2009;26:560-5
34. Olivieri NF, Saunthararajah Y, Thayalasuthan V, et al. A pilot study of subcutaneous decitabine in beta-thalassemia intermedia. Blood 2011;118:2708-11
35. Masera N, Tavecchia L, Capra M, et al. Optimal response to thalidomide in a patient with thalassaemia major resistant to conventional therapy. Blood Transfus 2010;8:63-5
36. Aguilar-Lopez LB, Delgado Lamas JL, Rubio-Jurado B, et al. Thalidomide therapy in a patient with thalassemia major. Blood Cells Mol Dis 2008;41:136-7
37. Perrine SP, Castaneda SA, Chui DH, et al. Fetal globin gene inducers: novel agents and new potential. Ann N Y Acad Sci 2010;1202:158-64
38. Perrine SP, Wargin WA, Boosalis MS, et al. Evaluation of safety and pharmacokinetics of sodium 2,2 dimethylbutyrate, a novel short chain fatty acid derivative, in a phase I, double-blind, placebo-controlled, single-dose, and repeat-dose studies in healthy volunteers. J Clin Pharmacol 2011;51:1186-94
39. Metallo CM, Azarin SM, Ji L, et al. Engineering tissue from human embryonic stem cells. J Cell Mol Med 2008;12:709-29
40. Li B, Xu L, Lu WY, et al. A whole-mechanical method to establish human embryonic stem cell line HN4 from discarded embryos. Cytotechnology 2010;62:509-18
41. Si-Tayeb K, Noto FK, Sepac A, et al. Generation of human induced pluripotent stem cells by simple transient transfection of plasmid DNA encoding reprogramming factors. BMC Dev Biol 2010;10:81-91
42. Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006;126:663-76 (Pubitemid 44233629)
43. Ye L, Chang JC, Lin C, et al. Induced pluripotent stem cells offer new approach to therapy in thalassemia and sickle cell anemia and option in prenatal diagnosis in genetic diseases. Proc Natl Acad Sci USA 2009;106:9826-30
44. Lozzio CB, Lozzio BB. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood 1975;45:321-34
45. Rutherford TR, Clegg JB, Weatherall DJ. K562 human leukaemic cells synthesise embryonic haemoglobin in response to haemin. Nature 1976;280:164-5 (Pubitemid 9221749)
46. Fibach E, Bianchi N, Borgatti M, et al. Effects of rapamycin on accumulation of alpha-, beta- and gamma-globin mRNAs in erythroid precursor cells from beta-thalassaemia patients. Eur J Haematol 2006;77:437-41 (Pubitemid 44483948)
47. Pope SH, Fibach E, Sun J, et al. Two-phase liquid culture system models normal human adult erythropoiesis at the molecular level. Eur J Haematol 2000;64:292-303 (Pubitemid 30307701)
48. Fibach E, Bianchi N, Borgatti M, et al. Mithramycin induces fetal hemoglobin production in normal and thalassemic human erythroid precursor cells. Blood 2003;102:1276-81 (Pubitemid 36988033)
49. Vadolas J, Nefedov M, Wardan H, et al. Humanized beta-thalassemia mouse model containing the common IVSI-110 splicing mutation. J Biol Chem 2006;281:7399-405 (Pubitemid 43847510)
50. Johnson RM, Gumucio D, Goodman M. Globin gene switching in primates. Comp Biochem Physiol A Mol Integr Physiol 2002;133:877-83 (Pubitemid 35351480)
51. Rivella S, May C, Chadburn A, et al. A novel murine model of Cooley anemia and its rescue by lentiviral-mediated human beta-globin gene transfer. Blood 2003;101:2932-9 (Pubitemid 36857977)
52. Weizer-Stern O, Adamsky K, Amariglio N, et al. mRNA expression of iron regulatory genes in beta-thalassemia intermedia and beta-thalassemia major mouse models. Am J Hematol 2006;81:479-83
53. Jamsai D, Zaibak F, Vadolas J, et al. A humanized BAC transgenic/knockout mouse model for HbE/beta-thalassemia. Genomics 2006;88:309-15 (Pubitemid 44278526)
54. Li W, Xie S, Guo X, et al. A novel transgenic mouse model produced from lentiviral germline integration for the study of beta-thalassemia gene therapy. Haematologica 2008;93:356-62 (Pubitemid 351398636)
55. Huo Y, McConnell SC, Ryan TM. Preclinical transfusion-dependent humanized mouse model of beta thalassemia major. Blood 2009;113:4763-70
56. Huo Y, McConnell SC, Liu S, et al. Humanized mouse models of Cooley's anemia: correct fetal-to-adult hemoglobin switching, disease onset, and disease pathology. Ann N Y Acad Sci 2010;1202:45-51
57. Bank A. On the road to gene therapy for beta-thalassemia and sickle cell anemia. Pediatr Hematol Oncol 2008;25:1-4 (Pubitemid 351168917)
58. Lisowski L, Sadelain M. Current status of globin gene therapy for the treatment of beta-thalassaemia. Br J Haematol 2008;141:335-45 (Pubitemid 351522768)
59. Breda L, Gambari R, Rivella S. Gene therapy in thalassemia and hemoglobinopathies. Mediterr J Hematol Infect Dis 2009;1:e2009008
60. Persons DA. Gene therapy: targeting beta-thalassaemia. Nature 2010;467:277-8
61. Bank A. Hemoglobin gene therapy for beta-thalassemia. Hematol Oncol Clin North Am 2010;24:1187-201
62. Boulad F, Riviere I, Sadelain M. Gene therapy for homozygous beta-thalassemia. Is it a reality? Hemoglobin 2009;33:S188-96
63. Yannaki E, Emery DW, Stamatoyannopoulos G. Gene therapy for beta-thalassaemia: the continuing challenge. Expert Rev Mol Med 2010;12:e31
64. Maina N, Zhong L, Li X, et al. Optimization of recombinant adeno-associated viral vectors for human beta-globin gene transfer and transgene expression. Hum Gene Ther 2008;19:365-75 (Pubitemid 351591432)
65. Gelinas R, Novak U. Retroviral vectors for the beta-globin gene that demonstrate improved titer and expression. Ann N y Acad Sci 1990;612:427-41
66. Dong WJ, Li B, Liu DP, et al. Evaluation of optimal expression cassette in retrovirus vector for beta-thalassemia gene therapy. Mol Biotechnol 2003;24:127-40 (Pubitemid 36623582)
67. Yi Y, Noh MJ, Lee KH. Current advances in retroviral gene therapy. Curr Gene Ther 2011;11:218-28
68. Sadelain M, Rivella S, Lisowski L, et al. Globin gene transfer for treatment of the beta-thalassemias and sickle cell disease. Best Pract Res Clin Haematol 2004;17:517-34 (Pubitemid 39462581)
69. Case SS, Price MA, Jordan CT, et al. Stable transduction of quiescent CD34 +CD38- human hematopoietic cells by HIV-1-based lentiviral vectors. Proc Natl Acad Sci USA 1999;96:2988-93 (Pubitemid 29148831)
70. Lisowski L, Sadelain M. Locus control region elements HS1 and HS4 enhance the therapeutic efficacy of globin gene transfer in beta-thalassemic mice. Blood 2007;110:4175-8 (Pubitemid 351377780)
71. Hargrove PW, Kepes S, Hanawa H, et al. Globin lentiviral vector insertions can perturb the expression of endogenous genes in beta-thalassemic hematopoietic cells. Mol Ther 2008;16:525-33 (Pubitemid 351314992)
72. Zhou HS, Zhao N, Li L, et al. Site-specific transfer of an intact beta-globin gene cluster through a new targeting vector. Biochem Biophys Res Commun 2007;356:32-7 (Pubitemid 46413124)
73. Zhao S, Weinreich MA, Ihara K, et al. In vivo selection of genetically modified erythroid cells using a jak2-based cell growth switch. Mol Ther 2004;10:456-68 (Pubitemid 39186109)
74. Persons DA. The challenge of obtaining therapeutic levels of genetically modified hematopoietic stem cells in beta-thalassemia patients. Ann N Y Acad Sci 2010;1202:69-74
75. Yannaki E, Stamatoyannopoulos G. Hematopoietic stem cell mobilization strategies for gene therapy of beta thalassemia and sickle cell disease. Ann N Y Acad Sci 2010;1202:59-63
76. Yannaki E, Psatha N, Athanasiou E, et al. Mobilization of hematopoietic stem cells in a thalassemic mouse model: implications for human gene therapy of thalassemia. Hum Gene Ther 2010;21:299-310
77. Miccio A, Cesari R, Lotti F, et al. In vivo selection of genetically modified erythroblastic progenitors leads to long-term correction of beta-thalassemia. Proc Natl Acad Sci USA 2008;105:10547-52
78. Persons DA, Allay ER, Sawai N, et al. Successful treatment of murine beta-thalassemia using in vivo selection of genetically modified, drug-resistant hematopoietic stem cells. Blood 2003;102:506-13 (Pubitemid 36841967)
79. Negre O, Fusil F, Colomb C, et al. Correction of murine beta-thalassemia after minimal lentiviral gene transfer and homeostatic in vivo erythroid expansion. Blood 2011;117:5321-31
80. Kumar P, Woon-Khiong C. Optimization of lentiviral vectors generation for biomedical and clinical research purposes: contemporary trends in technology development and applications. Curr Gene Ther 2011;11:144-53
81. Wilber A, Hargrove PW, Kim YS, et al. Therapeutic levels of fetal hemoglobin in erythroid progeny of beta-thalassemic CD34+ cells after lentiviral vector-mediated gene transfer. Blood 2011;117:2817-26
82. May C, Rivella S, Callegari J, et al. Therapeutic haemoglobin synthesis in beta-thalassaemic mice expressing lentivirus-encoded human beta-globin. Nature 2000;406:82-6 (Pubitemid 30460215)
83. May C, Rivella S, Chadburn A, Sadelain M. Successful treatment of murine beta-thalassemia intermedia by transfer of the human beta-globin gene. Blood 2002;99:1902-8 (Pubitemid 34525467)
84. Puthenveetil G, Scholes J, Carbonell D, et al. Successful correction of the human beta-thalassemia major phenotype using a lentiviral vector. Blood 2004;104:3445-53 (Pubitemid 39564412)
85. Imren S, Payen E, Westerman KA, et al. Permanent and panerythroid correction of murine beta thalassemia by multiple lentiviral integration in hematopoietic stem cells. Proc Natl Acad Sci USA 2002;99:14380-5 (Pubitemid 35257680)
86. Imren S, Fabry ME, Westerman KA, et al. High-level beta-globin expression and preferred intragenic integration after lentiviral transduction of human cord blood stem cells. J Clin Invest 2004;114:953-62 (Pubitemid 39578723)
87. Tisdale J, Sadelain M. Toward gene therapy for disorders of globin synthesis. Semin Hematol 2001;38:382-92 (Pubitemid 32995608)
88. Kafri T. Lentivirus vectors: difficulties and hopes before clinical trials. Curr Opin Mol Ther 2001;3:316-26 (Pubitemid 32755381)
89. Breda L, Kleinert DA, Casu C, et al. A preclinical approach for gene therapy of beta-thalassemia. Ann N Y Acad Sci 2010;1202:134-40
90. Bank A, Dorazio R, Leboulch P. A Phase I/II clinical trial of beta-globin gene therapy for beta-thalassemia. Ann N Y Acad Sci 2005;1054:308-16 (Pubitemid 43031038)
91. Arumugam P, Malik P. Genetic therapy for beta-thalassemia: from the bench to the bedside. Hematology Am Soc Hematol Educ Program 2010;2010:445-50
92. Sadelain M, Riviere I, Wang X, et al. Strategy for a multicenter phase I clinical trial to evaluate globin gene transfer in beta-thalassemia. Ann N Y Acad Sci 2010;1202:52-8
93. Cavazzana-Calvo M, Payen E, Negre O, et al. Transfusion independence and HMGA2 activation after gene therapy of human beta-thalassaemia. Nature 2010;467:318-22
94. Kaiser J. Gene therapy. beta-thalassemia treatment succeeds, with a caveat. Science 2009;326:1468-9
95. Colosimo A, Guida V, Antonucci I, et al. Sequence-specific modification of a beta-thalassemia locus by smallDNA fragments in human erythroid progenitor cells. Haematologica 2007;92:129-30 (Pubitemid 46232669)
96. Chin JY, Kuan JY, Lonkar PS, et al. Correction of a splice-site mutation in the beta-globin gene stimulated by triplex-forming peptide nucleic acids. Proc Natl Acad Sci USA 2008;105:13514-19
97. Rogers FA, Lin SS, Hegan DC, et al. Targeted gene modification of hematopoietic progenitor cells in mice following systemic administration of a PNA-peptide conjugate. Mol Ther 2012;20:109-18
98. Porteus MH. Mammalian gene targeting with designed zinc finger nucleases. Mol Ther 2006;13:438-46 (Pubitemid 43056910)
99. Zou J, Mali P, Huang X, et al. Site-specific gene correction of a point mutation in human iPS cells derived from an adult patient with sickle cell disease. Blood 2011;118:4599-608
100. Svasti S, Suwanmanee T, Fucharoen S, et al. RNA repair restores hemoglobin expression in IVS2-654 thalassemic mice. Proc Natl Acad Sci USA 2009;106:1205-10
101. El-Beshlawy A, Mostafa A, Youssry I, et al. Correction of aberrant pre-mRNA splicing by antisense oligonucleotides in beta-thalassemia Egyptian patients with IVSI-110 mutation. J Pediatr Hematol Oncol 2008;30:281-4 (Pubitemid 351507582)
102. Suwanmanee T, Sierakowska H, Fucharoen S, Kole R. Repair of a splicing defect in erythroid cells from patients with beta-thalassemia/HbE disorder. Mol Ther 2002;6:718-26 (Pubitemid 36054305)
103. Zeng Y, Gu X, Chen Y, et al. Reversal of aberrant splicing of beta-thalassemia allele by antisense RNA in vitro and in vivo. Chin Med J (Engl) 1999;112:107-11
104. El-Andaloussi S, Johansson HJ, Lundberg P, Langel U. Induction of splice correction by cell-penetrating peptide nucleic acids. J Gene Med 2006;8:1262-73 (Pubitemid 44660525)
105. Papapetrou EP, Lee G, Malani N, et al. Genomic safe harbors permit high beta-globin transgene expression in thalassemia induced pluripotent stem cells. Nat Biotechnol 2011;29:73-8
106. Sadelain M, Boulad F, Lisowki L, et al. Stem cell engineering for the treatment of severe hemoglobinopathies. Curr Mol Med 2008;8:690-7
107. Abuljadayel IS, Quereshi H, Ahsan T, et al. Infusion of autologous retrodifferentiated stem cells into patients with beta-thalassemia. ScientificWorldJournal 2006;6:1278-97 (Pubitemid 44715640)
108. Hanna J, Wernig M, Markoulaki S, et al. Treatment of sickle cell anemia mouse model with iPS cells generated from autologous skin. Science 2007;318:1920-3
109. Emery DW, Yannaki E, Tubb J, et al. Development of virus vectors for gene therapy of beta chain hemoglobinopathies: flanking with a chromatin insulator reduces gamma-globin gene silencing in vivo. Blood 2002;100:2012-19 (Pubitemid 35001231)
110. Hanawa H, Hargrove PW, Kepes S, et al. Extended beta-globin locus control region elements promote consistent therapeutic expression of a gamma-globin lentiviral vector in murine beta-thalassemia. Blood 2004;104:2281-90 (Pubitemid 39331824)
111. Persons DA, Hargrove PW, Allay ER, et al. The degree of phenotypic correction of murine beta-thalassemia intermedia following lentiviral-mediated transfer of a human gamma-globin gene is influenced by chromosomal position effects and vector copy number. Blood 2003;101:2175-83 (Pubitemid 36302055)
112. Hanawa H, Hargrove PW, Kepes S, et al. Extended beta-globin locus control region elements promote consistent therapeutic expression of a gamma-globin lentiviral vector in murine beta-thalassemia. Blood 2004;104:2281-90 (Pubitemid 39331824)
113. Nishino T, Tubb J, Emery DW. Partial correction of murine beta-thalassemia with a gammaretrovirus vector for human gamma-globin. Blood Cells Mol Dis 2006;37:1-7 (Pubitemid 44062363)
114. Nishino T, Cao H, Stamatoyannopoulos G, Emery DW. Effects of human gamma-globin in murine beta-thalassaemia. Br J Haematol 2006;134:100-8 (Pubitemid 43830035)
115. Thein SL, Menzel S. Discovering the genetics underlying foetal haemoglobin production in adults. Br J Haematol 2009;145:455-67
116. Forget BG. Progress in understanding the hemoglobin switch. N Engl J Med 2011;365:852-4
117. Sankaran VG, Xu J, Byron R, et al. A functional element necessary for fetal hemoglobin silencing. N Engl J Med 2011;365:807-14
118. Xu XS, Hong X, Wang G. Induction of endogenous gamma-globin gene expression with decoy oligonucleotide targeting Oct-1 transcription factor consensus sequence. J Hematol Oncol 2009;2:15-26
119. Jiang J, Best S, Menzel S, et al. cMYB is involved in the regulation of fetal hemoglobin production in adults. Blood 2006;108:1077-83 (Pubitemid 44154648)
120. Sankaran VG, Menne TF, Xu J, et al. Human fetal hemoglobin expression is regulated by the developmental stage-specific repressor BCL11A. Science 2008;322:1839-42
121. Sankaran VG, Xu J, Orkin SH. Transcriptional silencing of fetal hemoglobin by BCL11A. Ann N Y Acad Sci 2010;1202:64-8
122. Sankaran VG. Targeted therapeutic strategies for fetal hemoglobin induction. Hematology Am Soc Hematol Educ Program 2011;2011:459-65
123. Nguyen TK, Joly P, Bardel C, et al. The XmnIGgamma polymorphism influences hemoglobin F synthesis contrary to BCL11A and HBS1L-MYB SNPs in a cohort of 57 beta-thalassemia intermedia patients. Blood Cells Mol Dis 2010;45:124-7
124. Gabbianelli M, Morsilli O, Massa A, et al. Effective erythropoiesis and HbF reactivation induced by kit ligand in beta-thalassemia. Blood 2008;111:421-9
125. Borg J, Papadopoulos P, Georgitsi M, et al. Haploinsufficiency for the erythroid transcription factor KLF1 causes hereditary persistence of fetal hemoglobin. Nat Genet 2010;42:801-5
126. Bianchi N, Feriotto G, Gambari R, Mischiati C. Synthetic oligonucleotides as inducers of erythroid differentiation. PCTEP0102804; 2001
127. Graslund T, Li X, Magnenat L, et al. Exploring strategies for the design of artificial transcription factors: targeting sites proximal to known regulatory regions for the induction of gamma-globin expression and the treatment of sickle cell disease. J Biol Chem 2005;280:3707-14 (Pubitemid 40223838)
128. Wilber A, Tschulena U, Hargrove PW, et al. A zinc-finger transcriptional activator designed to interact with the gamma-globin gene promoters enhances fetal hemoglobin production in primary human adult erythroblasts. Blood 2010;115:3033-41
129. He L, Hannon GJ. MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet 2010;5:522-31
130. Krol J, Loedige I, Filipowicz W. The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet 2010;11:597-610
131. Sontheimer EJ, Carthew RW. Silence from within: endogenous siRNAs and miRNAs. Cell 2005;122:9-12 (Pubitemid 40977934)
132. Alvarez-Garcia I, Miska EA. MicroRNA functions in animal development and human disease. Development 2005;132:4653-62 (Pubitemid 41685276)
133. Sankaran VG, Menne TF, Scepanovic D, et al. MicroRNA-15a and -16-1 act via MYB to elevate fetal hemoglobin expression in human trisomy 13. Proc Natl Acad Sci USA 2001;108:1519-24
134. Voon HP, Vadolas J. Controlling alphaglobin: a review of alpha-globin expression and its impact on beta-thalassemia. Haematologica 2008;93:1868-76
135. Voon HP, Wardan H, Vadolas J. siRNA-mediated reduction of alpha-globin results in phenotypic improvements in beta-thalassemic cells. Haematologica 2008;93:1238-42
136. Xie SY, Ren ZR, Zhang JZ, et al. Restoration of the balanced alpha/betaglobin gene expression in beta654-thalassemia mice using combined RNAi and antisense RNA approach. Hum Mol Genet 2007;16:2616-25
137. Voon HP, Wardan H, Vadolas J. Co-inheritance of alpha- and beta-thalassaemia in mice ameliorates thalassaemic phenotype. Blood Cells Mol Dis 2007;39:184-8 (Pubitemid 47126925)
138. Fu YF, Du TT, Dong M, et al. Mir-144 selectively regulates embryonic alpha-hemoglobin synthesis during primitive erythropoiesis. Blood 2009;113:1340-9
139. Wang B, Fang Y, Guo X, et al. Transgenic human alpha-hemoglobin stabilizing protein could partially relieve betaIVS-2-654-thalassemia syndrome in model mice. Hum Gene Ther 2010;21:149-56
140. Free Patents Data Bank. Nottingham, MD, USA: FreePatentsOnline.com 2004 - 2012. Available from: http://www. freepatentsonline.com [Last accessed 14 February 2012]
141. Espacenet. Munich, Germany: European Patent Office. 2012. Available from: http://worldwide.espacenet.com/ [Last accessed 14 February 2012]
142. Clinical Trials Database. Bethesda, MD, USA: U.S. National Library of Medicine, 1993 - 2012. Available from: http:// clinicaltrials.gov/ [Last accessed 14 February 2012]
143. Sankaran VG, Nathan DG. Thalassemia: an overview of 50 years of clinical research. Hematol Oncol Clin North Am 2010;24:1005-20
144. Salvatori F, Breveglieri G, Zuccato C, et al. Production of beta-globin and adult hemoglobin following G418 treatment of erythroid precursor cells from homozygous beta039 thalassemia patients. Am J Hematol 2009;84:720-8
145. Guideline on quality, non-clinical and clinical aspects of medicinal products containing genetically modified cells. London, UK: European medicines Agency, May 20, 2010. Available from: www.ema.europa.eu/ema/pages/includes/ document/open-document.jsp?webContentId=WC500093309 [Last accessed 14 February 2012]
146. Gambari R and Finotti A. Bioethics and freedom of scientific research in gene therapy and stem cell biology. In: Bin R, Lorenzon S, Lucchi N, editors. Biotech Innovations and Fundamental Rights. Springer; Heildelberg: 2012. p. 115-30
147. Wilber A, Hargrove PW, Kim YS, et al. Therapeutic levels of fetal hemoglobin in erythroid progeny of beta-thalassemic CD34+ cells after lentiviral vector-mediated gene transfer. Blood 2011;117:2817-26
148. Katada H, Komiyama M. Artificial restriction DNA cutters to promote homologous recombination in human cells. Curr Gene Ther 2011;11:38-45