Skip to main content
SpringerLink
Log in

BRAF V600E mutation detection in hairy cell leukemia-utility of archival DNA from bone marrow aspirate/imprint smear and amplification refractory mutation system

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

BRAF V600E is a disease defining mutation for hairy cell leukemia (HCL), which helps in its diagnosis and differentiation from morphologically similar splenic marginal zone lymphoma (SMZL) and HCL-variant (HCL-v). Forty eight cases:HCL(n = 34), SMZL(n = 11) and HCL-v(n = 3) were included. Of these, 32 were retrospective and 16 were prospective. DNA was extracted, in retrospective cases from cells obtained by smears from bone marrow aspirate/trephine imprint (BMA/BMTx) slides, and in prospective cases from peripheral blood (PB)/BMA samples. BRAF V600E mutation testing was done using ARMS-PCR. BRAF V600E mutation was positive in all HCL and negative in all SMZL and HCL-v cases. DNA extracted from BMA/BMTx slides gave results comparable to DNA extracted from PB/BMA samples. Median age of presentation for HCL (53 years) and SMZL (56 years) were quite similar, however, HCL-v (71 years) cases presented at an older age. Statistically significant differences between the three groups were seen for total leucocyte, platelet, absolute lymphocyte and monocyte counts, bone marrow-infiltration pattern, reticulin fibrosis, and an expression of CD11c, CD25, CD103, CD123, and CD200. The use of BMA/BMTx smears for DNA extraction was found to be a useful alternative to DNA extraction from formalin-fixed paraffin-embedded biopsy sections. ARMS-PCR is an efficient and specific technique to detect BRAF V600E mutation in HCL patients.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Foucar K, Falini B, Stein H (2017) Hairy cell leukaemia. In: Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H, et al. (eds) WHO classification of tumours of haematopoietic and lymphoid tissues, 4th edn. Lyon, IARC, pp 226–228

    Google Scholar 

  2. Sharpe RW, Bethel KJ (2006) Hairy cell leukemia: diagnostic pathology. Hematol Oncol Clin North Am 20:1023–1049

    Article  Google Scholar 

  3. Bethel KJ, Sharpe RW (2003) Pathology of hairy cell leukaemia. Best Pract Res Clin Haematol 16:15–31

    Article  CAS  Google Scholar 

  4. Matutes E (2006) Immunophenotyping and differential diagnosis of hairy cell leukemia. Hematol Oncol Clin N Am 20:1051–1063

    Article  Google Scholar 

  5. Shao H, Calvo KR, Gronborg M, Tembhare PR, Kreitman RJ, Steller-Stevenson M et al (2013) Distinguishing hairy cell leukemia variant from hairy cell leukemia: development and validation of diagnostic criteria. Leuk Res 37:401–409

    Article  CAS  Google Scholar 

  6. Gidron A, Tallman MS (2005) Hairy cell leukemia: towards a curative strategy. Hematol Oncol Clin N Am 20:1153–1162

    Article  Google Scholar 

  7. Else M, Ruchlemer R, Osuji N et al (2005) Long remissions in hairy cell leukemia with purine analogs: a report of 219 patients with a median follow-up of 12.5 years. Cancer 104:2442–2448

    Article  CAS  Google Scholar 

  8. Tiacci E, Trifonov V, Schiavoni G et al (2011) BRAF mutations in hairy-cell leukemia. N Engl J Med 364:2305–2315

    Article  CAS  Google Scholar 

  9. Wan PT, Garnett MJ, Roe SM, Lee S, Niculescu-Duvaz D, Good VM et al (2004) Mechanism of activation of the RAF-ERK signaling pathway by oncogenic mutations of B-RAF. Cell 116(6):855–867

    Article  CAS  Google Scholar 

  10. Tiacci E, Schiavoni G, Forconi F, Santi A, Trentin L, Ambrosetti A et al (2012) Simple genetic diagnosis of hairy cell leukemia by sensitive detection of the BRAF-V600E mutation. Blood 119(1):192–195

    Article  CAS  Google Scholar 

  11. Verma S, Greaves WO, Ravandi F, Reddy N, Bueso-Ramos CE, O’Brien S et al (2012) Rapid detection and quantitation of BRAF mutations in hairy cell leukemia using a sensitive pyrosequencing assay. Am J Clin Pathol 138(1):153–156

    Article  CAS  Google Scholar 

  12. Chen D, Wang YY, Chuai ZR, Huang JF, Wang YX, Liu K et al (2014) High-Resolution melting analysis for accurate detection of BRAF mutations: a systematic review and meta-analysis. Sci Rep 4:4168

    Article  Google Scholar 

  13. Carbonell P, Turpin MC, Torres-Moreno D, Molina-Martinez I, García-Solano J, Perez-Guillermo M et al (2011) Comparison of allelic discrimination by dHPLC, HRM, and TaqMan in the detection of BRAF mutation V600E. J Mol Diagn 13(5):467–473

    Article  CAS  Google Scholar 

  14. Caper D, Preusser M, Habel A, Sahm F, Ackermann U, Schindler G et al (2011) Assessment of BRAF V600E mutation status by immunohistochemistry with a mutation-specific monoclonal antibody. Acta Neuropathol 122(1):11–19

    Article  Google Scholar 

  15. Foucar K, Falini B, Stein H (2017) Hairy cell leukaemia. In: Swerdlow SH, Campo E, Harris NL, et al (eds) WHO classification of tumours of haematopoietic and lymphoid tissues. Lyon, IARC, pp 226–228

    Google Scholar 

  16. Piris M, BMA, Issacson PG, Swerldow PG, et al (2017) Splenic marginal zone lymphoma. In: Swerdlow SH, Campo E, Harris NL, et al (eds) WHO classification of tumours of haematopoietic and lymphoid tissues. Lyon, IARC, pp 223–225

    Google Scholar 

  17. Piris MA, PittalugaFoucar K, Mollejo M, et al (2017) Hairy cell leukaemia variant. In: Swerdlow SH, Campo E, Harris NL, et al (eds) WHO classification of tumours of haematopoietic and lymphoid tissues. IARC, Lyon, pp 230–231

    Google Scholar 

  18. Huang T, Zhuge J, Zhang WW (2013) Sensitive detection of BRAF V600E mutation by amplification refractory mutation system (ARMS)-PCR. Biomark Res 1:3

    Article  Google Scholar 

  19. World Health Organization (2001) Dept. of Nutrition for Health and Development. Iron deficiency anaemia : assessment, prevention and control : a guide for programme managers. World Health Organization, Geneva

    Google Scholar 

  20. Bates I, Lewis SM (2011) Reference ranges and normal values. In: Bain BJ, Bates I, Laffan MA, Lewis SM (eds) Practical haematology, 11th edn. Churchill Livingstone, Philadelphia, pp 11–22

    Google Scholar 

  21. Stasi R (2012) How to approach thrombocytopenia. Hematol Am Soc Hematol Educ Program 2012:191–197

    Article  Google Scholar 

  22. Grever MR (2010) How I treat hairy cell leukemia. Blood 115:21–28

    Article  CAS  Google Scholar 

  23. Grever MR, Abdel-Wahab O, Andritsos LA, Banerji V, Barrientos J, Blachly J et al (2017) Consensus guidelines for the diagnosis and management of patients with classic hairy cell leukemia. Blood 129:553–560

    Article  CAS  Google Scholar 

  24. Arcaini L, Zibellini S, Boveri E, Riboni R, Rattotti S, Varettoni M et al (2012) The BRAF V600E mutation in hairy cell leukemia and other mature B-cell neoplasms. Blood 119:188–191

    Article  CAS  Google Scholar 

  25. Blombery PA, Wong SQ, Hewitt CA, Dobrovic A, Maxwell EL, Juneja S et al (2012) Detection of BRAF mutations in patients with hairy cell leukemia and related lymphoproliferative disorders. Haematologica 97:780–783

    Article  CAS  Google Scholar 

  26. Samuel J, Macip S, Dyer MJS (2014) Efficacy of vemurafenib in hairy-cell leukemia. N Engl J Med 370:286–288

    Article  CAS  Google Scholar 

  27. Tiacci E, Park JH, Carolis LD, Chung SS, Broccoli A, Scott S et al (2015) Targeting mutant BRAF in relapsed or refractory Hairy-Cell Leukemia. N Engl J Med 373:1733–1747

    Article  CAS  Google Scholar 

  28. Xi L, Arons E, Navarro W, Calvo KR, Stetler-Stevenson M, Raffeld M et al (2012) Both variant and IGHV4-34-expressing hairy cell leukemia lack the BRAF V600E mutation. Blood 119:3330–3332

    Article  CAS  Google Scholar 

  29. Thomas C, Amanuel B, Finlayson J, Grieu- Iacopetta F, Spagnolo DV, Erber WN (2015) BRAF mutation detection in hairy cell leukaemia from archival haematolymphoid specimens. Pathology 47:349–354

    Article  CAS  Google Scholar 

  30. Dores GM, Matsuno RK, Weisenburger DD, Rosenberg PS, Anderson WF (2008) Hairy cell leukaemia: a heterogeneous disease? Br J Hematol 142:45–51

    Article  Google Scholar 

  31. Teras LR, DeSantis CE, Cerhan JR, Morton LM, Jemal A, Flowers CR (2016) 2016 US lymphoid malignancy statistics by World Health Organization subtypes. CA Cancer J Clin 66:443–459

    Article  Google Scholar 

  32. Galani KS, Subramanian PG, Gadage VS, Rahman K, Ashok Kumar MS, Shinde S et al (2012) Clinico-pathological profile of Hairy cell leukemia: critical insights gained at a tertiary care cancer hospital. Indian J Pathol Microbiol 55:61–65

    Article  Google Scholar 

  33. Yuan CM, Yang LJ (2008) Hairy cell leukemia with unusual loss of CD103 in a subset of the neoplastic population: immunophenotypic and cell cycle analysis by flow cytometry. Int J Clin Exp Pathol 1(4):381–386

    PubMed  PubMed Central  Google Scholar 

  34. Chen YH, Tallman MS, Goolsby C, Peterson L (2006) Immunophenotypic variations in hairy cell leukemia. Am J Clin Pathol 125(2):251–259

    Article  Google Scholar 

Download references

Acknowledgements

This study was supported in part by “Intramural Research Grant (No. 71/8-Edu-15/176) to Dr. Shano Naseem, from Postgraduate Institute of Medical Education and Research, Chandigarh, India”.

Author information

Authors and Affiliations

  1. Department of Hematology, Postgraduate Institute of Medical Education and Research, Chandigarh, 160012, India

    Shano Naseem, Ojas Gupta, Jogeshwar Binota & Neelam Varma

  2. Department of Internal Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India

    Subhash Varma & Pankaj Malhotra

Authors
  1. Shano Naseem
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ojas Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jogeshwar Binota
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Neelam Varma
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Subhash Varma
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Pankaj Malhotra
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Shano Naseem.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Naseem, S., Gupta, O., Binota, J. et al. BRAF V600E mutation detection in hairy cell leukemia-utility of archival DNA from bone marrow aspirate/imprint smear and amplification refractory mutation system. Mol Biol Rep 47, 4365–4372 (2020). https://doi.org/10.1007/s11033-020-05509-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-020-05509-0

Keywords

Search

Navigation