Abstract
Co-inheritance of gamma and beta globin gene mutations in a compound heterozygous state is rare but of clinical interest as it provides an important data on understanding the HbF expression. Hematological analysis was carried out (Sysmex KX-21). F-cells were enumerated using flow cytometry. Beta globin gene was analysed by CRDB technique and by DNA sequencing. Gamma globin promoter region was sequenced and expression studies were carried out using real time Taqman assay. We report a family, where two inherited defects of the β globin gene cluster segregate. The proband and her sibling were compound heterozygotes for a novel Gγ promoter mutation and the 619 bp deletion a common Indian β thalassemia mutation. Molecular characterization revealed that the father (HbA2 5.1%, HbF 5.4%), proband (HbA2 3.6%, HbF 31.7%) and her brother (HbA2 3.9%, HbF 23.6%) were heterozygous for the 619 bp deletion. The mother (HbA2 2.1%, HbF 3.4%) had a normal β globin gene. As both the children showed high HbF levels, the γ globin gene work up was carried out. The Gγ-globin gene promoter analysis revealed that the mother and the two children were heterozygous for a 5 bp deletion -ATAAG (-533 to -529) that resides in the GATA binding site. These findings suggest that the 5 bp deletion in the Gγ globin promoter has a functional role in silencing the γ-globin gene expression in adults by disrupting GATA-1 binding and the associated repressor complex and results in the up-regulation of gamma globin gene expression. When co-inherited with β -thalassemia trait it leads to a phenotype of HPFH.
This is a preview of subscription content, log in via an institution to check access.
References
Langdon S, Kaufman R (1998) Gamma-globin gene promoter elements required for interaction with globin enhancers. Blood 91:309–318
Katsantoni EZ, Langeveld A, Wai A et al (2003) Persistent γ-globin expression in adult transgenic mice is mediated by HPFH-2, HPFH-3, and HPFH-6 breakpoint sequences. Blood 1021:3412–3419
Tasiopoulou M, Boussiou M, Sinopoulou K et al (2008) Gγ-196 C→T, Aγ-201 C→T: two novel mutations in the promoter region of γ-globin genes associated with non deletional hereditary persistence of fetal hemoglobin in Greece. Blood Cells Mol Dis 40:320–322
Huang XD, Yang XO, Huang R et al (2000) A Novel four base-pair deletion within the Ag globin gene promoter associated with slight increase of Ag expression in adult. Am J Hematol 63:16–19
Hoyer J, Penz C, Virgil F et al (2002) Diagnostic usefulness in the distinction of hereditary persistence of fetal hemoglobin (HPFH) and hemoglobin S–HPFH from other conditions with elevated levels of hemoglobin F. Am J Clin Pathol 117:857–863
Italia KY, Colah R, Mohanty D (2006) Evaluation of F cells in sickle cell disorders by flow cytometry –comparison with the Kleihauer–Betke’s slide method. Int J Lab Hematol 29:409–414
Singleton B, Burton N, Green C et al (2008) Mutations in EKLF/KLF1 form the molecular basis of the rare blood group In(Lu) phenotype. Blood 112:2081–2088
Xu XS, Hong X, Wang G (2009) Induction of endogenous γ-globin gene expression with decoy oligonucleotide targeting Oct-1 transcription factor consensus sequence. J Hematol Oncol 2:1–11
Mastropietro F, Modiano G, Cappabianca MP et al (2002) Factors regulating Hb F synthesis in thalassemic diseases. BMC Blood Disord 2:1–7
Chassanidis C, Kalamaras A, Phylactides M et al (2009) The Hellenic type of nondeletional hereditary persistence of fetal hemoglobin results from a novel mutation (g.-109G>T) in the gene promoter. Ann Hematol 88:549–555
Papadakis MN, Patrinos GP, Tsaftaridis P et al (2002) A comparative study of Greek nondeletional hereditary persistence of fetal hemoglobin and β thalassemia compound heterozygotes. J Mol Med 80:243–247
Dedoussis GVZ, Sinopoulou K, Gyparaki M et al (1999) Fetal hemoglobin expression in the compound heterozygous state for -117 (G > A) AgγHPFH and IVSII-745 (C > G) β + thalassemia: a case study. Am J Hematol 61:139–143
Chen Z, Luo H, Basran RK et al (2008) A T-to-G transversion at nucleotide _567 upstream of HBG2 in GATA-1 binding motif is associated with elevated hemoglobin F. Mol Cell Biol 28:4386–4393
Zaker-Kandjani B, Namdar-Aligoodarzi P, Azarkeivan A et al (2015) Mutation screening of the Kruppel-like factor 1 gene using single-strand conformational polymorphism in a cohort of Iranian β-thalassemia patients. Hemoglobin 39:24–29
Sankaran VG (2011) Targeted therapeutic strategies for fetal hemoglobin induction. Hematology 459–465
Tallack M, Perkins A (2013) Three fingers on the switch: Kruppel-like factor 1 regulation of γ-globin to β-globin gene switching. Curr Opin Hematol 20:193–200
Waye JS, Eng B (2015) Kruppel-like factor 1: hematologic phenotypes associated with KLF1 gene mutations. Int J Lab Hematol 37:78–84
Gallienne AE, Dréau HMP, Schuh A et al (2012) Ten novel mutations in the erythroid transcription factor KLF1 gene associated with increased fetal hemoglobin levels in adults. Haematologica 97:430–434
De Souza Carrocini G, Zamaro PA et al (2011) What influences Hb fetal production in adulthood? Rev Bras Hematol Hemoter 33:231–236
Sawant M, Chandrakala S, Colah R et al (2016) Does HbF induction by hydroxycarbamide work through MIR210 in sickle cell anaemia patients? Br J Haematol 173:786–803
Wienert B, Funnell A, Norton L et al (2015) Editing the genome to introduce a beneficial naturally occurring mutation associated with increased fetal globin. Nat Commun 6:1–8
Acknowledgements
This work was funded by Indian Council of Medical Research (ICMR), New Delhi, India. We thank the family for participating in the study. We also thank University of Mumbai for their support.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Rights and permissions
About this article
Cite this article
Hariharan, P., Sawant, M., Gorivale, M. et al. Synergistic effect of two β globin gene cluster mutations leading to the hereditary persistence of fetal hemoglobin (HPFH) phenotype. Mol Biol Rep 44, 413–417 (2017). https://doi.org/10.1007/s11033-017-4125-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-017-4125-0