F8 Gene Splice Donor Mutation (c.1271+1G>A) in Individual with Mild Hemophilia A in Indonesia: A Case Study

Kirana, Ni Gusti Ayu Galuh Candra; Suprianto, Suprianto; Lesmana, Indra; Sukorini, Usi; Handayani, Niken Satuti Nur

doi:10.61186/ijbc.16.4.1

Volume 16, Issue 4 (December 2024 2024) Iranian Journal of Blood and Cancer 2024, 16(4): 1-8 | Back to browse issues page

‎ 10.61186/ijbc.16.4.1

Mendeley

Zotero

RefWorks

Kirana N G A G C, Suprianto S, Lesmana I, Sukorini U, Handayani N S N. F8 Gene Splice Donor Mutation (c.1271+1G>A) in Individual with Mild Hemophilia A in Indonesia: A Case Study. Iranian Journal of Blood and Cancer 2024; 16 (4) :1-8
URL: http://ijbc.ir/article-1-1577-en.html

F8 Gene Splice Donor Mutation (c.1271+1G>A) in Individual with Mild Hemophilia A in Indonesia: A Case Study

Ni Gusti Ayu Galuh Candra Kirana¹

, Suprianto Suprianto¹

, Indra Lesmana²

, Usi Sukorini³

, Niken Satuti Nur Handayani ^*⁴

1- Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia
2- Department of Tropical Biology, Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia. & Laboratory of Genetics and Breeding, Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia.
3- Department of Clinical Pathology and Laboratory Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia
4- Department of Tropical Biology, Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia. & Laboratory of Genetics and Breeding, Faculty of Biology, Universitas Gadjah Mada, Yogyakarta, Indonesia. , niken_satuti@ugm.ac.id

Abstract: (356 Views)

Introduction: Hemophilia A is a bleeding disorder caused by a deficiency of coagulation factor VIII. Hemophilia A is an X-linked recessive disorder. Depending on the level of blood coagulation factor VIII, hemophilia severity is classified as mild (5-40%), moderate (1-5%), or severe (<1%). The absence of hemophilia A mutation studies in Indonesia makes this topic important to study.
Methods: This study detected and classified F8 gene mutations. A member of the Indonesian Hemophilia Society Association for the Special Region of Yogyakarta provided saliva for DNA testing. Long-read sequencing data were performed using the next-generation sequencing (NGS) technique via the Oxford Nanopore Technologies plc (ONT) PromethION 24 platform. The mutation was confirmed using Sanger sequencing, after amplifying intron 8 of the F8 gene with the PCR technique. The F8 gene intron 8 nucleotide sequence was aligned using the alignment tool on the Benchling website.
Results: The results of this study showed that there was a splice donor site mutation in intron 8 of the F8 gene (c.1271+1G>A) in one patient. This mutation can cause the occurrence of cryptic splice donor sites. Cryptic splice donor site prediction was carried out using the splice donor prediction tool available on the NNSPLICE website. The appearance of cryptic splice donor sites can lead to the formation of out-of-frame proteins.
Conclusions: The F8 gene mutation causing hemophilia A was detected using long-read sequencing and the next-generation sequencing (NGS) technique. The type of mutation identified is a splice donor site mutation, specifically the variant c.1271+1G>A, in sample code HM13.

Keywords: Hemophilia A, F8 gene, Intron 8, Donor splice mutation, c.1271+1G

Full-Text [PDF 581 kb] (165 Downloads)

: Original Article | Subject: Genetics
Received: 2024/10/26 | Accepted: 2024/12/24 | Published: 2024/12/30

References

1. Choi SJ, Jang KJ, Lim JA, Kim HS. Human coagulation factor VIII domain-specific recombinant polypeptide expression. Blood Res. 2015;50(2):103-8. [DOI:10.5045/br.2015.50.2.103]

2. Shih MY, Wang J Der, Yin J De, Tsan YT, Chan WC. Differences in major bleeding events between Patients with severe hemophilia A and hemophilia B: A Nationwide, population-based cohort study. Clin Appl Thromb. 2019;25:1-7. [DOI:10.1177/1076029619888023]

3. Jayakrishnan T, Shah D, Mewawalla P. Hemophilia C: A case report with updates on diagnosis and management of a rare bleeding disorder. J Hematol. 2019;8(3):144-7. [DOI:10.14740/jh522]

4. Alkarrash MS, Badawi R, Sallah H, Shashaa MN, Argilo J, Alkhoury R. Hemophilia A and C in a female: The first case report in literature. Annals of Medicine and Surgery. 2021; 68:1-3. [DOI:10.1016/j.amsu.2021.102561]

5. Goodeve AC. Hemophilia B: Molecular Pathogenesis and mutation analysis. J Thromb Haemost. 2015;13(7):1184-95. [DOI:10.1111/jth.12958]

6. Miller CH. The clinical genetics of hemophilia B (Factor IX Deficiency). Appl Clin Genet. 2021;14: 445-454. [DOI:10.2147/TACG.S288256]

7. Mandal S, Gami S, Shah S. A case report on an extremely rare disease: Factor XI deficiency. Cureus. 2020;12(10):1-4. [DOI:10.7759/cureus.10746]

8. Berntorp E, Fischer K, Hart DP, Mancuso ME, Stephensen D, Shapiro AD, et al. Haemophilia. Nat Rev Dis Prim. 2021;7(45):1-19. [DOI:10.1038/s41572-021-00278-x]

9. Zarrilli F, Coppola A, Schiavulli M, Cimino E, Elce A, Rescigno G, et al. Haemophilia a: The consequences of de novo mutations. Two case reports. Blood Transfus. 2018;16(4):392-3.

10. Shen BW, Spiegel PC, Chang CH, Huh JW, Lee JS, Kim J, et al. The tertiary structure and domain organization of coagulation factor VIII. Blood. 2008;111(3):1240-1247. [DOI:10.1182/blood-2007-08-109918]

11. Bowen DJ. Haemophilia A and haemophilia B: Molecular insights. J Clin Pathol: Mol Pathol. 2002;55(2):127-144. https://doi.org/10.1136/mp.55.1.1 [DOI:10.1136/mp.55.2.127]

12. Zahari M, Sulaiman SA, Othman Z, Ayob Y, Karim FA, Jamal R. Mutational profiles of F8 and F9 in a cohort of haemophilia A and haemophilia B patients in the multi-ethnic Malaysian population. Mediterr J Hematol Infect Dis. 2018;10(1):1-13. [DOI:10.4084/mjhid.2018.056]

13. Atik T, Işık E, Onay H, Akgün B, Shamsali M, Kavaklo K, et al. Factor 8 gene mutation spectrum of 270 patients with hemophilia A: Identification of 36 novel mutations. Turkish Journal of Haematology. 2020;37(3):145-153. [DOI:10.4274/tjh.galenos.2020.2019.0262]

14. Shinozawa K, Amano K, Hagiwara T, Bingo M, Chikasawa Y, Inaba H, et al. Genetic analysis of carrier status in female members of Japanese hemophilia families. Journal of Thrombosis and Haemostasis. 2021;19(6):1493-1505. [DOI:10.1111/jth.15301]

15. Elisa Bach J, Wolf B, Oldenburg J, Muller CR, Rost S. Identification of deep intronic variants in 15 haemophilia A patients by next generation sequencing of the whole factor VIII gene. Thromb Haemost. 2015;114(4):757-67. [DOI:10.1160/TH14-12-1011]

16. Uen C, Oldenburg J, Schroder J, Brackmann H J, Schwaab R, Schneppenheim R, et al. Hamophilie A patienten ohne mutation im FVIII-Gen [2% haemophilia A patients without mutation in the FVIII gene]. Hamostaseologie. 2003;1-5. [DOI:10.1055/s-0037-1619567]

17. World Federation of Hemophilia. World federation of hemophilia report on the annual global survey 2021. World Fed Hemoph. 2021;1-100.

18. Wang J, Gu J, Chen H, Wu Q, Xiong L, Qiao B, et al. A novel deletion mutation of the F8 gene for hemophilia A. Diagnostics. 2022;21;12(11):2876. [DOI:10.3390/diagnostics12112876]

19. Sinclair A. Genetic 101 : Detecting mutation in human genes. CMAJ. 2002;167(3):275-279.

20. Graw, J., Brackmann, H. H., Oldenburg, J., Schneppenheim, R., Spannagl, M., and Schwaab, R. 2005. Haemophilia A: from Mutation Analysis to new Therapies. Nature Reviews Genetics. 6(6):488-501. [DOI:10.1038/nrg1617]

21. Pshenichnikova, O., Salomashkina, V., Poznyakova, J., Selivanova, D., Chernetskaya, D., Yakovleva, E., Dimitrieva, O., Likhacheva, E., Perina, F., Zozulya, N., & Surin, V. (2023). Spectrum of Causative Mutations in Patients with Hemophilia A in Russia. Genes, 14(2), 260. [DOI:10.3390/genes14020260]

22. Santacroce, R., Acquila, M., Belvini, D., Castaldo, G., Garagiola, I., Giacomelli, S. H., Lombardi, A. M., Minuti, B., Riccardi, F., Salviato, R., Tagliabue, L., Grandone, E., Margaglione, M., & AICE-Genetics Study Group (2008). Identification of 217 unreported mutations in the F8 gene in a group of 1,410 unselected Italian patients with hemophilia A. Journal of human genetics, 53(3), 275-284. [DOI:10.1007/s10038-007-0238-y]

23. Akkarapatumwong, V., Intorasoot, S., Oranwiroon, S., Thano-Otarakul, P., Pung-Amritt, P., Veerakul, G., Mahasandana, C., Panyim, S., & Yenchitsomanus, P. (2000). Frameshift mutations with severe and moderate clinical phenotypes in Thai hemophilia A patients. Human mutation. 16(6):1-4. https://doi.org/10.1002/1098-1004(200012)16:6<530::AID-HUMU16>3.0.CO;2-3 [DOI:10.1002/1098-1004(200012)16:63.0.CO;2-3]

24. Akkarapatumwong, V., Oranwiroon, S., Pung-amritt, P., Treesucon, A., Thanootarakul, P., Veerakul, G., Mahasandana, C., Panyim, S. and Yenchitsomanus, P. (1999). Mutations of the factor VIII gene in Thai hemophilia A patients. Human mutation. 15: 1-6. https://doi.org/10.1002/(SICI)1098-1004(200001)15:1<117::AID-HUMU27>3.0.CO;2-E [DOI:10.1002/(SICI)1098-1004(200001)15:13.0.CO;2-E]

25. Guo, Z., Yang, L., Qin, X., Liu, X., & Zhang, Y. (2018). Spectrum of Molecular Defects in 216 Chinese Families With Hemophilia A: Identification of Noninversion Mutation Hot Spots and42 Novel Mutations. journal of the International Academy of Clinical and Applied Thrombosis/Hemostasis, 24(1): 70-78. [DOI:10.1177/1076029616687848]

26. Khanum, F., Collins, P. W., Harris, R. L., & Bowen, D. J. (2014). Characterization of F8 defects in haemophilia A in Pakistan: investigation of correlation between mutation type and the in vitro thrombin generation assay. Haemophilia : the official journal of the World Federation of Hemophilia, 20(2), 287-293. [DOI:10.1111/hae.12272]

27. Johnsen, J. M., Fletcher, S. N., Huston, H., Roberge, S., Martin, B. K., Kircher, M., Josephson, N. C., Shendure, J., Ruuska, S., Koerper, M. A., Morales, J., Pierce, G. F., Aschman, D. J., & Konkle, B. A. (2017). Novel approach to genetic analysis and results in 3000 hemophilia patients enrolled in the My Life, Our Future initiative. Blood advances, 1(13), 824-834. [DOI:10.1182/bloodadvances.2016002923]

28. Liu, W., Lyu, C., Wang, W., Xue, F., Chen, L., Li, H., Chi, Y., Ma, Y., Wu, R., Fang, Y., Zhang, L., & Yang, R. (2022). Risk Factors For Inhibitors In Hemophilia A based on RNA-seq and DNA Methylation. Research And Practice In Thrombosis And Haemostasis. 6(6), e12794. [DOI:10.1002/rth2.12794]

29. Jankowska, K. I., McGill, J., Pezeshkpoor, B., Oldenburg, J., Sauna, Z. E., & Atreya, C. D. (2020). Further Evidence That MicroRNAs Can Play a Role in Hemophilia A Disease Manifestation: F8 Gene Downregulation by miR-19b-3p and miR-186-5p. Frontiers In Cell And Developmental Biology. 8, 669. [DOI:10.3389/fcell.2020.00669]

30. Cutler JA, Mitchell MJ, Smith MP, Savidge GF. The identification and classification of 41 novel mutations in the factor VIII gene (F8C). Hum Mutat. 2002;19(3):274-8. [DOI:10.1002/humu.10056]

31. Jackson LJ. A reappraisal of non-consensus mRNA splice sites. Nucleic Acids Res. 1991;19(14):3795-8. [DOI:10.1093/nar/19.14.3795]

32. Caminsky N, Mucaki EJ, Rogan PK. Interpretation of mRNA splicing mutations in genetic disease: Review of the literature and guidelines for information-theoretical analysis. F1000 Research. 2014;3(282):1-30. https://doi.org/10.12688/f1000research.5654.2 [DOI:10.12688/f1000research.5654.1]

33. Anna A, Monika G. Splicing mutations in human genetic disorders: Examples, detection, and confirmation. J Appl Genet. 2018;59(3):253-68. [DOI:10.1007/s13353-018-0444-7]

34. Nguyen H, Das U, Wang B, Xie J. The matrices and constraints of GT/AG splice sites of more than 1000 Species/Lineages. Gene. 2018;660:92-101. [DOI:10.1016/j.gene.2018.03.031]

35. Zimmermann MA, Gehrig A, Oldenburg J, Muller CR., Rost S. Analysis of F8 mRNA in haemophilia A patients with silent mutations or presumptive splice site mutations. Haemophilia. 2013;19(2):310-317. [DOI:10.1111/hae.12039]

36. Baralle FE, Singh RN, Stamm S. RNA structure and splicing regulation. Physiol Behav. 2017;176(3):139-148.

37. Rogalska ME, Vivori C, Valcárcel J. Regulation of pre-mRNA splicing: roles in physiology and disease, and therapeutic prospects. Nat Rev Genet. 2023;24(4):251-69. [DOI:10.1038/s41576-022-00556-8]

38. Balestra D, Maestri I, Branchini A, Ferrarese M, Bernardi F, Pinotti M. An altered splicing registry explains the differential ExSpeU1-mediated rescue of splicing mutations causing haemophilia A. Front Genet. 2019;10:1-9. [DOI:10.3389/fgene.2019.00974]

39. Aldalaqan S, Dalgliesh C, Luzzi S, Siachisumo C, Reynard LN, Ehrmann I, et al. Cryptic splicing: Common pathological mechanisms involved in male infertility and neuronal diseases. Cell Cycle. 2022;21(3):219-27. [DOI:10.1080/15384101.2021.2015672]

40. Kapustin Y, Chan E, Sarkar R, Wong F, Vorechovsky I, Winston RM, et al. Cryptic splice sites and split genes. Nucleic Acids Res. 2011;39(14):5837-44. [DOI:10.1093/nar/gkr203]

41. Lannoy N, Abinet I, Bosmans A, Lambert C, Vermylen C, Hermans C. Computational and molecular approaches for predicting unreported causal missense mutations in Belgian patients with haemophilia A. Haemophilia. 2012;18(3):1-9. [DOI:10.1111/j.1365-2516.2011.02640.x]

42. Castaman G, Giacomelli SH, Mancuso ME, Sanna S, Santagostino E, Rodeghiero F. F8 mRNA studies in Haemophilia A patients with different splice site mutations. Haemophilia. 2010;16(5):786-90. [DOI:10.1111/j.1365-2516.2010.02250.x]

43. Wang J, Ye Z, Huang TH, Shi H, Jin VX. Computational methods and correlation of exon-skipping events with splicing, transcription, and epigenetic factors. Methods Mol Biol. 2017;1513:163-70. [DOI:10.1007/978-1-4939-6539-7_11]

44. Lee Y, Gamazon ER, Rebman E, Lee Y, Lee S, Dolan ME, et al. Variants affecting exon skipping contribute to complex traits. PLoS Genet. 2012;8(10):1-12. [DOI:10.1371/journal.pgen.1002998]

45. Marian AJ. Clinical interpretation and management of genetic variants. JACC Basic to Transl Sci. 2020;5(10):1029-42. [DOI:10.1016/j.jacbts.2020.05.013]

46. Bagnall, R. D., Waseem, N. H., Green, P. M., Colvin, B., Lee, C., & Giannelli, F. (1999). Creation of a novel donor splice site in intron 1 of the factor VIII gene leads to activation of a 191 bp cryptic exon in two haemophilia A patients. British journal of haematology, 107(4), 766-771. [DOI:10.1046/j.1365-2141.1999.01767.x]

47. Dardik R, Janczar S, Lalezari S, Avishai E, Levy-Mendelovich S, Barg AA, et al. Four decades of carrier detection and prenatal diagnosis in hemophilia A: historical overview, state of the art and future directions. Int J Mol Sci. 2023;24(14):1-13. [DOI:10.3390/ijms241411846]

48. Kawashima S, Hattori A, Suzuki E, Matsubara K, Toki M, Kosaki R, et al. Methylation status of genes escaping from X-chromosome inactivation in patients with X-chromosome rearrangements. Clin Epigenetics. 2021;13(1):1-11. [DOI:10.1186/s13148-021-01121-6]

49. Miller CH, Benson J, Ellingsen D, Driggers J, Payne A, Kelly FM, et al. F8 and F9 mutations in US haemophilia patients: correlation with history of inhibitor and race/ethnicity. Haemophilia. 2012;18(3):375-82. [DOI:10.1111/j.1365-2516.2011.02700.x]

50. Doncel SS, Alejandra G, Cortes JM, Rico CA, Javier F, Cadavid M, et al. Haemophilia A : A review of clinical manifestations, treatment, mutations, and the development of inhibitors pathways FVIIIa. 2023;15(1):130-150. [DOI:10.3390/hematolrep15010014]

51. Luo L, Zheng Q, Chen Z, Huang M, Fu L, Hu J, et al. Hemophilia a patients with inhibitors: Mechanistic insights and novel therapeutic implications. Front Immunol. 2022;13:1-15. [DOI:10.3389/fimmu.2022.1019275]

Send email to the article author

Rights and permissions
	This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Designed & Developed by : Yektaweb