MOLECULAR ANALYSIS OF NON-TRANSFUSION DEPENDENT THALASSEMIA ASSOCIATED WITH HEMOGLOBIN E-?-THALASSEMIA DISEASE WITHOUT ?--THALASSEMIA
Main Article Content
Keywords
Non-transfusion dependent thalassemia; HBS1L-MYB gene; BCL11A gene; KLF1 gene; Gγ-XmnI polymorphism
Abstract
Background: The finding of many Thai Hb E-?0-thalassemia patients with non-transfusion dependent thalassemia (NTDT) phenotype without co-inheritance of ?-thalassemia has prompted us to investigate the existence of other genetic modifying factors.
Methods: Study was done on 146 adult Thai patients with NTDT Hb E-?0-thalassemia and a homozygous ?-thalassemia patient without co-inheritance of ?-thalassemia. Multiple single-nucleotide polymorphisms (SNPs) associated with ?-globin gene expression including the G?-XmnI of HBG2 gene, rs2297339, rs4895441, and rs9399137 of the HBS1L-MYB gene, rs4671393 in the BCL11A gene, and G176AfsX179, T334R, R238H and -154 (C-T) in the KLF1 gene were investigated using PCR-and related techniques.
Results: Heterozygous and homozygous for Gg-XmnI of HBG2 gene were detected at 68.0% and 6.1%, respectively. Further DNA analysis identified the rs2297339 (C-T), rs4895441 (A-G), and rs9399137 (T-C) of HBS1L-MYB gene in 86.4%, 22.5% and 20.4%, respectively. The rs4671393 (G-A) of the BCL11A gene was found at 31.3%. For the KLF1 gene, the T334R and G176AfsX179 (+/-) were detected at 8.2% and 1.4%, respectively.
Conclusion: It was found that these SNPs when analyzed in combination could explain the mild phenotypic expression of all cases. These results underline the importance of these informative SNPs on phenotypic expression of Hb E-?-thalassemia patients.
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https://doi.org/10.1179/1607845414Y.0000000211
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in northeast Thailand: The E-SAAN study phase I. Hematology
2018;23:55-60.
https://doi.org/10.1080/10245332.2017.1358845
PMid:28759343
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PMid:23729725 PMCid:PMC3669437
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amelioration of the phenotype of beta-thalassemia. Proc Natl Acad Sci
USA 2008;105:1620-1625.
https://doi.org/10.1073/pnas.0711566105
PMid:18245381 PMCid:PMC2234194
23. Wahlberg K, Jiang J, Rooks H, et al. The HBS1L-MYB intergenic
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https://doi.org/10.1182/blood-2009-03-210146
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mutations in the erythroid transcription factor KLF1 gene associated
with increased fetal hemoglobin levels in adults. Haematologica
2012;97:340-343.
https://doi.org/10.3324/haematol.2011.055442
PMid:22102705 PMCid:PMC3291586
25. Thein SL. Molecular basis of ?-thalassemia and potential therapeutic
targets. Blood Cells Mol Dis 2018;70:54-65.
https://doi.org/10.1016/j.bcmd.2017.06.001
PMid:28651846 PMCid:PMC5738298
26. Rani N, Jamwal M, Kaur J, et al. Homozygous KLF1 mutation
c.901C>T (p.Arg301Cys) resulting in mild thalassemia intermedia in an
Indian: A next-generation sequencing diagnosis. Blood Cells Mol Dis
2018;72:19-21.
https://doi.org/10.1016/j.bcmd.2018.06.003
PMid:29980343
27. Chaouch L, Moumni I, Ouragini H, et al. rs11886868 and rs4671393 of
BCL11A associated with Hb F level variation and modulate clinical
events among sickle cell anemia patients. Hematology 2016;27:425-429.
https://doi.org/10.1080/10245332.2015.1107275
PMid:27077760
28. Nguyen TK, Joly P, Bardel C, Moulsma M, Bonello-Palot N, Francina
A. The XmnI (G)gamma polymorphism influences hemoglobin F
synthesis contrary to BCL11A and HBS1L-MYB SNPs in a cohort of 57
beta-thalassemia intermedia patient. Blood Cells Mol Dis 2010;45:124-
127.
https://doi.org/10.1016/j.bcmd.2010.04.002
PMid:20472475
Blackwell Science; 2001.
https://doi.org/10.1002/9780470696705
2. Yamsri S, Sanchaisuriya K, Fucharoen G, Sae-Ung N, Ratanasiri T,
Fucharoen S. Prevention of severe thalassemia in northeast Thailand: 16
years of experience at a single university center. Prenat Diagn
2010;30:540-546.
https://doi.org/10.1002/pd.2514
PMid:20509153
3. Italia K, Dabke P, Sawant P, Nadkarni A, Ghosh K, Colah RB. Hb E-?thalassemia in five Indian states. Hemoglobin 2016;40:310-315.
https://doi.org/10.1080/03630269.2016.1201487
PMid:27623935
4. George E, Wong HB. Hb E beta +-thalassaemia in West Malaysia:
clinical features in the most common beta-thalassaemia mutation of the
Malays [IVS 1-5 (G-->C)], Singapore Med J 1993;34:500-503.
5. Winichagoon P, Fucharoen S, Chen P, Wasi P. Genetic factors affecting
clinical severity in beta-thalassemia syndromes. J Pediatr Hematol
Oncol 2000;22:573-580.
https://doi.org/10.1097/00043426-200011000-00026
PMid:11132233
6. Fucharoen S, Ketvichit P, Pootrakul P, Siritanaratkul N, Piankijagum A,
Wasi P. Clinical manifestation of beta-thalassemia/hemoglobin E
disease. J Pediatr Hematol Oncol 2000;22:552-557.
www.mjhid.org Mediterr J Hematol Infect Dis 2019; 11; e2019038 Pag. 7 / 7
https://doi.org/10.1097/00043426-200011000-00022
PMid:11132229
7. Sleiman J, Tarhini A, Bou-Fakhredin R, Saliba AN, Cappellini MD,
Taher AT. Non-transfusion dependent thalassemia: an update on
complications and management. Int J Mol Sci 2018;19:182.
https://doi.org/10.3390/ijms19010182
PMid:29316681 PMCid:PMC5796131
8. Nuntakarn L, Fucharoen S, Fucharoen G, Sanchaisuriya K, Jetsrisuparb
A, Wiangnon S. Molecular, hematological and clinical aspects of
thalassemia major and thalassemia intermedia associated with Hb Ebeta-thalassemia in northeast Thailand. Blood Cells Mol Dis
2009;42:32-35.
https://doi.org/10.1016/j.bcmd.2008.09.002
PMid:18951049
9. Yamsri S, Pakdee N, Fucharoen G, Sanchaisuriya K, Fucharoen S.
Molecular understanding of non-transfusion-dependent thalassemia
associated with hemoglobin E-beta-thalassemia in northeast Thailand.
Acta Haematol 2016;136:233-239.
https://doi.org/10.1159/000449120
PMid:27710960
10. Nuinoon M, Makarasara W, Mushiroda T, et al. A genome-wide
association identified the common genetic variants influence disease
severity in ?0-thalassemia/hemoglobin E. Hum Genet 2010;127:303-314.
https://doi.org/10.1007/s00439-009-0770-2
PMid:20183929
11. Lettre G, Sankaran VG, Bezerra MA, et al. DNA polymorphisms at the
BCL11A, HBS1L-MYB, and beta-globin loci associate with fetal
hemoglobin levels and pain crises in sickle cell disease. Proc Natl Acad
Sci USA 2008;105:11869-11874.
https://doi.org/10.1073/pnas.0804799105
PMid:18667698 PMCid:PMC2491485
12. Sedgewick AE, Timofeev N, Sebastiani P, et al. BCL11A is a major Hb
F quantitative trait locus in three different populations with betahemoglobinopathies. Blood Cells Mol Dis 2008;41:255-258.
https://doi.org/10.1016/j.bcmd.2008.06.007
PMid:18691915 PMCid:PMC4100606
13. Creary LE, Ulug P, Menzel S, et al. Genetic variation on chromosome 6
influences F cell levels in healthy individuals of African descent and Hb
F levels in sickle cell patients. PLoS One 2009;4:e4218.
https://doi.org/10.1371/journal.pone.0004218
PMid:19148297 PMCid:PMC2621086
14. So CC, Song YQ, Tsang ST, et al. The HBS1L-MYB intergenic region
on chromosome 6q23 is a quantitative trait locus controlling fetal
haemoglobin level in carriers of beta-thalassaemia. J Med Genet
2008;45:745-751.
https://doi.org/10.1136/jmg.2008.060335
PMid:18697826
15. Pakdee N, Yamsri S, Fucharoen G, Sanchaisuriya K, Pissard S,
Fucharoen S. Variability of hemoglobin F expression in hemoglobin EE
disease: hematological and molecular analysis. Blood Cells Mol Dis
2014;53:11-15.
https://doi.org/10.1016/j.bcmd.2014.02.005
PMid:24581976
16. Tepakhan W, Yamsri S, Fucharoen G, Sanchaisuriya K, S. Fucharoen S.
Krüppel-like factor 1 mutations and expression of hemoglobins F and
A2 in homozygous hemoglobin E syndrome. Ann Hematol
2015;94:1093-1098.
https://doi.org/10.1007/s00277-015-2335-x
PMid:25694242
17. Tepakhan W, Yamsri S, Sanchaisuriya K, Fucharoen G, Xu X,
Fucharoen S. Nine known and five novel mutations in the erythroid
transcription factor KLF1 gene and phenotypic expression of fetal
hemoglobin in hemoglobin E disorder. Blood Cells Mol Dis 2016;59:85-
91.
https://doi.org/10.1016/j.bcmd.2016.04.010
PMid:27282573
18. Prayalaw P, Teawtrakul N, Jetsrisuparb A, Pongudom S, G. Fucharoen S.
Non transfusion-dependent thalassemia in northeast Thailand. Acta
Haematol 2016;135:15-20.
https://doi.org/10.1159/000435802
PMid:26303193
19. Teawtrakul N, Pussadhamma B, Ungprasert P, et al. A risk score for
predicting pulmonary hypertension in patients with non-transfusion
dependent thalassemia in northeast Thailand: The E-SAAN score.
Hematology 2015;20:416-420.
https://doi.org/10.1179/1607845414Y.0000000211
PMid:25386747
20. Teawtrakul N, Jetsrisuparb A, Pongudom S, et al. Epidemiologic study
of major complications in adolescent and adult patients with thalassemia
in northeast Thailand: The E-SAAN study phase I. Hematology
2018;23:55-60.
https://doi.org/10.1080/10245332.2017.1358845
PMid:28759343
21. Musallam KM, Rivella S, Vichinsky E, Rachmilewitz EA. Nontransfusion-dependent thalassemias. Haematologica 2013;98:833-844.
https://doi.org/10.3324/haematol.2012.066845
PMid:23729725 PMCid:PMC3669437
22. Uda M, Galanello R, Sanna S, et al. Genome-wide association study
shows BCL11A associated with persistent fetal hemoglobin and
amelioration of the phenotype of beta-thalassemia. Proc Natl Acad Sci
USA 2008;105:1620-1625.
https://doi.org/10.1073/pnas.0711566105
PMid:18245381 PMCid:PMC2234194
23. Wahlberg K, Jiang J, Rooks H, et al. The HBS1L-MYB intergenic
interval associated with elevated Hb F levels shows characteristics of a
distal regulatory region in erythroid cells. Blood 2009;114:1254-1262.
https://doi.org/10.1182/blood-2009-03-210146
PMid:19528534
24. Gallienne AE, Dreau HM, Schuh A, Old JM, Henderson S. Ten novel
mutations in the erythroid transcription factor KLF1 gene associated
with increased fetal hemoglobin levels in adults. Haematologica
2012;97:340-343.
https://doi.org/10.3324/haematol.2011.055442
PMid:22102705 PMCid:PMC3291586
25. Thein SL. Molecular basis of ?-thalassemia and potential therapeutic
targets. Blood Cells Mol Dis 2018;70:54-65.
https://doi.org/10.1016/j.bcmd.2017.06.001
PMid:28651846 PMCid:PMC5738298
26. Rani N, Jamwal M, Kaur J, et al. Homozygous KLF1 mutation
c.901C>T (p.Arg301Cys) resulting in mild thalassemia intermedia in an
Indian: A next-generation sequencing diagnosis. Blood Cells Mol Dis
2018;72:19-21.
https://doi.org/10.1016/j.bcmd.2018.06.003
PMid:29980343
27. Chaouch L, Moumni I, Ouragini H, et al. rs11886868 and rs4671393 of
BCL11A associated with Hb F level variation and modulate clinical
events among sickle cell anemia patients. Hematology 2016;27:425-429.
https://doi.org/10.1080/10245332.2015.1107275
PMid:27077760
28. Nguyen TK, Joly P, Bardel C, Moulsma M, Bonello-Palot N, Francina
A. The XmnI (G)gamma polymorphism influences hemoglobin F
synthesis contrary to BCL11A and HBS1L-MYB SNPs in a cohort of 57
beta-thalassemia intermedia patient. Blood Cells Mol Dis 2010;45:124-
127.
https://doi.org/10.1016/j.bcmd.2010.04.002
PMid:20472475