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Genetic variation of Krüppel-like factor 1 (KLF1) and fetal hemoglobin (HbF) levels in β0-thalassemia/HbE disease

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Abstract

Heterogeneity of HbF levels in β0-thalassemia/HbE disease has been reported to be associated with variations in clinical manifestations of the disease, and several genetic-modifying factors beyond the β-globin gene cluster have been identified as HbF regulators. Down-regulation or heterozygous mutations of Krüppel-like factor 1 (KLF1) is associated with elevated HbF levels in non-thalassemia subjects. This study confirms that experimental down-regulation of KLF1 in β0-thalassemia/HbE-derived erythroblasts significantly increases HbF production (up to 52.3 ± 2.4%), albeit with slightly delayed erythroid terminal differentiation. KLF1 exome sequencing of 130 Thai β0-thalassemia/HbE patients without co-inheritance of α-thalassemia found six patients with KLF1 heterozygous mutations including rs2072596 (p.F182L; n = 5) and rs745347362 (p.P284L; n = 1) missense mutations. However, while these patients had high HbF levels (38.1 ± 7.5%), they were all associated with a severe clinical phenotype. These results suggest that while reduction of KLF1 expression in β0-thalassemia/HbE erythroblasts can increase HbF levels, it is not sufficient to alleviate the clinical phenotype.

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References

  1. Weatherall DJ, Clegg JB. The thalassemia syndromes. Oxford: Blackwell Science; 2001.

    Book  Google Scholar 

  2. Fucharoen S, Winichagoon P. Haemoglobinopathies in southeast Asia. Indian J Med Res. 2011;134:498–506.

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Sripichai O, Fucharoen S. Fetal hemoglobin regulation in beta-thalassemia: heterogeneity, modifiers and therapeutic approaches. Expert Rev Hematol. 2016;9:1129–37. doi:10.1080/17474086.2016.1255142.

    Article  CAS  PubMed  Google Scholar 

  4. Nuinoon M, Makarasara W, Mushiroda T, Setianingsih I, Wahidiyat PA, Sripichai O, et al. A genome-wide association identified the common genetic variants influence disease severity in β0-thalassemia/hemoglobin E. Hum Genet. 2010;127:303–14. doi:10.1007/s00439-009-0770-2.

    Article  CAS  PubMed  Google Scholar 

  5. Borg J, Papadopoulos P, Georgitsi M, Gutierrez L, Grech G, Fanis P, et al. Haploinsufficiency for the erythroid transcription factor KLF1 causes hereditary persistence of fetal hemoglobin. Nat Genet. 2010;42:801–5. doi:10.1038/ng.630.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Zhou D, Liu K, Sun CW, Pawlik KM, Townes TM. KLF1 regulates BCL11A expression and gamma- to beta-globin gene switching. Nat Genet. 2010;42:742–4. doi:10.1038/ng.637.

    Article  CAS  PubMed  Google Scholar 

  7. Liu D, Zhang X, Yu L, Cai R, Ma X, Zheng C, et al. KLF1 mutations are relatively more common in a thalassemia endemic region and ameliorate the severity of beta-thalassemia. Blood. 2014;124:803–11. doi:10.1182/blood-2014-03-561779.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Viprakasit V, Ekwattanakit S, Riolueang S, Chalaow N, Fisher C, Lower K, et al. Mutations in Kruppel-like factor 1 cause transfusion-dependent hemolytic anemia and persistence of embryonic globin gene expression. Blood. 2014;123:1586–95. doi:10.1182/blood-2013-09-526087.

    Article  CAS  PubMed  Google Scholar 

  9. Yu LH, Liu D, Cai R, Shang X, Zhang XH, Ma XX, et al. Changes in hematological parameters in alpha-thalassemia individuals co-inherited with erythroid Kruppel-like factor mutations. Clin Genet. 2015;88:56–61. doi:10.1111/cge.12443.

    Article  CAS  PubMed  Google Scholar 

  10. Tepakhan W, Yamsri S, Fucharoen G, Sanchaisuriya K, Fucharoen S. Kruppel-like factor 1 mutations and expression of hemoglobins F and A2 in homozygous hemoglobin E syndrome. Ann Hematol. 2015;94:1093–8. doi:10.1007/s00277-015-2335-x.

    Article  CAS  PubMed  Google Scholar 

  11. Magor GW, Tallack MR, Gillinder KR, Bell CC, McCallum N, Williams B, et al. KLF1-null neonates display hydrops fetalis and a deranged erythroid transcriptome. Blood. 2015;125:2405–17. doi:10.1182/blood-2014-08-590968.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Sripichai O, Makarasara W, Munkongdee T, Kumkhaek C, Nuchprayoon I, Chuansumrit A, et al. A scoring system for the classification of beta-thalassemia/Hb E disease severity. Am J Hematol. 2008;83:482–4.

    Article  PubMed  Google Scholar 

  13. Bhanu NV, Trice TA, Lee YT, Gantt NM, Oneal P, Schwartz JD, et al. A sustained and pancellular reversal of gamma-globin gene silencing in adult human erythroid precursor cells. Blood. 2005;105:387–93. doi:10.1182/blood-2004-04-1599.

    Article  CAS  PubMed  Google Scholar 

  14. Palii CG, Pasha R, Brand M. Lentiviral-mediated knockdown during ex vivo erythropoiesis of human hematopoietic stem cells. J Vis Exp. 2011;. doi:10.3791/2813.

    PubMed  PubMed Central  Google Scholar 

  15. Fucharoen S, Shimizu K, Fukumaki Y. A novel C-T transition within the distal CCAAT motif of the G gamma-globin gene in the Japanese HPFH: implication of factor binding in elevated fetal globin expression. Nucleic Acids Res. 1990;18:5245–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Barrett JC, Fry B, Maller J, Daly MJ. Haploview: analysis and visualization of LD and haplotype maps. Bioinformatics. 2005;21:263–5. doi:10.1093/bioinformatics/bth457.

    Article  CAS  PubMed  Google Scholar 

  17. Gnanapragasam MN, Bieker JJ. Orchestration of late events in erythropoiesis by KLF1/EKLF. Curr Opin Hematol. 2017;. doi:10.1097/MOH.0000000000000327.

    PubMed  Google Scholar 

  18. Gnanapragasam MN, McGrath KE, Catherman S, Xue L, Palis J, Bieker JJ. EKLF/KLF1-regulated cell cycle exit is essential for erythroblast enucleation. Blood. 2016;128:1631–41. doi:10.1182/blood-2016-03-706671.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Arnaud L, Saison C, Helias V, Lucien N, Steschenko D, Giarratana MC, et al. A dominant mutation in the gene encoding the erythroid transcription factor KLF1 causes a congenital dyserythropoietic anemia. Am J Hum Genet. 2010;87:721–7. doi:10.1016/j.ajhg.2010.10.010.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Tallack MR, Keys JR, Humbert PO, Perkins AC. EKLF/KLF1 controls cell cycle entry via direct regulation of E2f2. J Biol Chem. 2009;284:20966–74. doi:10.1074/jbc.M109.006346.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Perseu L, Satta S, Moi P, Demartis FR, Manunza L, Sollaino MC, et al. KLF1 gene mutations cause borderline HbA(2). Blood. 2011;118:4454–8. doi:10.1182/blood-2011-04-345736.

    Article  CAS  PubMed  Google Scholar 

  22. Drissen R, Palstra RJ, Gillemans N, Splinter E, Grosveld F, Philipsen S, et al. The active spatial organization of the beta-globin locus requires the transcription factor EKLF. Genes Dev. 2004;18:2485–90. doi:10.1101/gad.317004.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Donze D, Jeancake PH, Townes TM. Activation of delta-globin gene expression by erythroid Krupple-like factor: a potential approach for gene therapy of sickle cell disease. Blood. 1996;88:4051–7.

    CAS  PubMed  Google Scholar 

  24. Tang DC, Ebb D, Hardison RC, Rodgers GP. Restoration of the CCAAT box or insertion of the CACCC motif activates [corrected] delta-globin gene expression. Blood. 1997;90:421–7.

    CAS  PubMed  Google Scholar 

  25. Xu J, Sankaran VG, Ni M, Menne TF, Puram RV, Kim W, et al. Transcriptional silencing of {gamma}-globin by BCL11A involves long-range interactions and cooperation with SOX6. Genes Dev. 2010;24:783–98. doi:10.1101/gad.1897310.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Libani IV, Guy EC, Melchiori L, Schiro R, Ramos P, Breda L, et al. Decreased differentiation of erythroid cells exacerbates ineffective erythropoiesis in beta-thalassemia. Blood. 2008;112(3):875–85. doi:10.1182/blood-2007-12-126938.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Siatecka M, Bieker JJ. The multifunctional role of EKLF/KLF1 during erythropoiesis. Blood. 2011;118:2044–54. doi:10.1182/blood-2011-03-331371.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Helias V, Saison C, Peyrard T, Vera E, Prehu C, Cartron JP, et al. Molecular analysis of the rare in(Lu) blood type: toward decoding the phenotypic outcome of haploinsufficiency for the transcription factor KLF1. Hum Mutat. 2013;34:221–8. doi:10.1002/humu.22218.

    Article  CAS  PubMed  Google Scholar 

  29. Ngo D, Bae H, Steinberg MH, Sebastiani P, Solovieff N, Baldwin CT, et al. Fetal hemoglobin in sickle cell anemia: genetic studies of the Arab–Indian haplotype. Blood Cells Mol Dis. 2013;51:22–6. doi:10.1016/j.bcmd.2012.12.005.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Radmilovic M, Zukic B, Petrovic MS, Bartsakoulia M, Stankovic B, Kotur N, et al. Functional analysis of a novel KLF1 gene promoter variation associated with hereditary persistence of fetal hemoglobin. Ann Hematol. 2013;92:53–8. doi:10.1007/s00277-012-1625-9.

    Article  CAS  PubMed  Google Scholar 

  31. Zaker-Kandjani B, Namdar-Aligoodarzi P, Azarkeivan A, Najmabadi H, Banan M. Mutation screening of the Kruppel-like factor 1 gene using single-strand conformational polymorphism in a cohort of Iranian beta-thalassemia patients. Hemoglobin. 2015;39:24–9. doi:10.3109/03630269.2014.991023.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by grants from Mahidol University and by a National Science and Technology Development Agency (NSTDA) Research Chair Grant. PK is supported by a Thailand Graduate Institute of Science and Technology (TGIST) scholarship (TG-22-14-58-037D) from the National Science and Technology Development Agency (NSTDA), Thailand. We sincerely thank all donors who participated in this study.

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Authors and Affiliations

  1. Institute of Molecular Biosciences, Mahidol University, Salaya Campus, 25/25 Phuttamonthon Sai 4, Salaya, Nakhon Pathom, Thailand

    Pinyaphat Khamphikham, Orapan Sripichai, Thongperm Munkongdee, Suthat Fucharoen & Duncan R. Smith

  2. National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani, Thailand

    Sissades Tongsima

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  1. Pinyaphat Khamphikham
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Correspondence to Duncan R. Smith.

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Khamphikham, P., Sripichai, O., Munkongdee, T. et al. Genetic variation of Krüppel-like factor 1 (KLF1) and fetal hemoglobin (HbF) levels in β0-thalassemia/HbE disease. Int J Hematol 107, 297–310 (2018). https://doi.org/10.1007/s12185-017-2357-3

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  • DOI: https://doi.org/10.1007/s12185-017-2357-3

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