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Molecular and clinical significance of FLT3, NPM1, DNMT3A and TP53 mutations in acute myeloid leukemia patients

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Abstract

Background

Acute myeloid leukemia (AML) is a type of blood cancer that affects the bone marrow and blood cells. AML is characterized by the rapid growth and accumulation of abnormal white blood cells, known as myeloblasts, which interfere with the production of normal blood cells.

Aims

The main aim was to determine the relationship between these genetic alterations and the clinico-haematological parameters and prognostic factors with therapy for Iraqi patients with AML.

Methods

We used Sanger Sequencing to detect the mutations in 76 AML patients. Clinical data of AML patients were retrospectively analysed to compare the prognosis of each gene mutation group.

Results

Somatic mutations were identified in 47.4% of the enrolled patients in a core set of pathogenic genes, including FLT3 (18 patients, 23.7%), DNMT3A (14, 18.4%), NPM1 (11, 14.5%) and TP53 (5, 6.8%). As multiple mutations frequently coexisted in the same patient, we classified patients into 10 further groups. Two novel mutations were detected in FLT3-ITD, with new accession numbers deposited into NCBI GenBank (OP807465 and OP807466). These two novel mutations were computationally analysed and predicted as disease-causing mutations. We found significant differences between patients with and without the detected mutations in disease progression after induction therapy (remission, failure and death; pv = < 0.001) and statistically significant differences were reported in total leukocyte count (pv = < 0.0001).

Conclusion

These genes are among the most frequently mutated genes in AML patients. Understanding the molecular and clinical significance of these mutations is important for guiding treatment decisions and predicting patient outcomes.

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Data Availability

The datasets regarding FLT3-ITD gene generated during the study were deposited into the NCBI GenBank Banklt database and can be accessed with the accession numbers OP807465 and OP807466 (https://www.ncbi.nlm.nih.gov/nuccore/OP807465/https://www.ncbi.nlm.nih.gov/nuccore/OP807466). Furthermore, the data related to DNMT3A gene in our study released as: (https://www.ncbi.nlm.nih.gov/nuccore/ON881282.1) (https://www.ncbi.nlm.nih.gov/nuccore/ON881281.1) and (https://www.ncbi.nlm.nih.gov/nuccore/ON881280.1).

Code Availability

Not Applicable.

References

  1. Arber DA et al (2016) The 2016 revision to the World Health Organization classification of myeloid neoplasms and acute leukemia. Blood 127:2391–2405

    Article  CAS  PubMed  Google Scholar 

  2. Tang S et al (2017) FLT3-ITD with DNMT3A R882 double mutation is a poor prognostic factor in chinese patients with acute myeloid leukemia after chemotherapy or allogeneic hematopoietic stem cell transplantation. Int J Hematol 106:552–561

    Article  CAS  PubMed  Google Scholar 

  3. Loghavi S et al (2014) Clinical features of de Novo acute myeloid leukemia with concurrent DNMT3A, FLT3 and NPM1 mutations. J Hematol Oncol 7:1–10

    Article  Google Scholar 

  4. Papaemmanuil E et al (2016) Genomic classification and prognosis in Acute myeloid leukemia. N Engl J Med 374:2209–2221

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Volpe G et al (2015) Regulation of the flt3 gene in haematopoietic stem and early progenitor cells. PLoS ONE 10:1–18

    Article  Google Scholar 

  6. Schlenk RF et al (2014) Differential impact of allelic ratio and insertion site in FLT3-ITD-positive AML with respect to allogeneic transplantation. Blood 124:3441–3449

    Article  CAS  PubMed  Google Scholar 

  7. Verson CA (2020) Profiling FLT3 mutations in mexican Acute Myeloid Leukemia Pediatric Patients: impact on overall survival. 8,

  8. Bhattacharyya J et al (2018) Prevalence and clinical significance of FLT3 and NPM1 mutations in Acute myeloid leukaemia patients of Assam, India. Indian J Hematol Blood Transfus 34:32–42

    Article  PubMed  Google Scholar 

  9. George B, Kantarjian H, Baran N, Krocker JD, Rios A (2021) Tp53 in acute myeloid leukemia: molecular aspects and patterns of mutation. Int J Mol Sci 22,

  10. Huang X et al (2013) A novel PTEN/Mutant p53/ c-Myc/Bcl-XL axis mediates context-dependent oncogenic effects of PTEN with implications for cancer prognosis and therapy. Neoplasia (United States) 15:952–965

    Article  Google Scholar 

  11. Hou H et al (2015) TP53 mutations in de novo acute myeloid leukemia patients: longitudinal follow-ups show the mutation is stable during disease evolution. https://doi.org/10.1038/bcj.2015.59

  12. DiNardo C, Lachowiez C (2019) Acute myeloid leukemia: from mutation profiling to treatment decisions. Curr Hematol Malig Rep 14:386–394

    Article  PubMed  Google Scholar 

  13. Genomic and Epigenomic Landscapes of Adult De Novo Acute Myeloid Leukemia (2013) N Engl J Med 368:2059–2074

    Article  Google Scholar 

  14. Bullinger L, Konstanze D (2022) Genomics of Acute Myeloid Leukemia Diagnosis and Pathways 35,

  15. Lazenby M et al (2014) The prognostic relevance of flt3 and npm1 mutations on older patients treated intensively or non-intensively: a study of 1312 patients in the UK NCRI AML16 trial. Leukemia 28:1953–1959

    Article  CAS  PubMed  Google Scholar 

  16. Döhner H et al (2022) Diagnosis and management of AML in adults: 2022 recommendations from an international expert panel on behalf of the ELN. Blood 140:1345–1377

    Article  PubMed  Google Scholar 

  17. Stein EM et al (2021) Safety and Efficacy of Menin Inhibition in patients (pts) with MLL-Rearranged and NPM1 mutant Acute Leukemia: a phase (ph) 1, first-in-human study of SNDX-5613 (AUGMENT 101). Blood 138:699–699

    Article  Google Scholar 

  18. Uy GL et al (2021) Flotetuzumab as salvage immunotherapy for refractory acute myeloid leukemia. Blood 137:751–762

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Untergasser A et al (2007) Primer3Plus, an enhanced web interface to Primer3. Nucleic Acids Res 35:71–74

    Article  Google Scholar 

  20. Kalendar R, Khassenov B, Ramankulov Y, Samuilova O, Ivanov K (2017) I. FastPCR: an in silico tool for fast primer and probe design and advanced sequence analysis. Genomics 109:312–319

    Article  CAS  PubMed  Google Scholar 

  21. Manachai N et al (2022) Activating Mutation in the Receptor Tyrosine Kinase FLT3 with Clinicopathological Relevance in Canine Mast Cell Tumors. Vet. Med. Int (2022)

  22. Park DJ et al (2020) Characteristics of DNMT3A mutations in acute myeloid leukemia. 55,

  23. Gamaleldin MA, Imbaby SA, E. (2021) T. The role of tumor necrosis factor receptor superfamily member 4 (TNFRSF4) gene expression in diagnosis and prognosis of acute myeloid leukemia. Mol Biol Rep 48:6831–6843

    Article  CAS  PubMed  Google Scholar 

  24. Kadia TM et al (2016) TP53 mutations in newly diagnosed acute myeloid leukemia: clinicomolecular characteristics, response to therapy, and outcomes. Cancer 122:3484–3491

    Article  CAS  PubMed  Google Scholar 

  25. Ali AM, Salih GF (2022) Identification of three novel DNMT3A mutations with compromising methylation capacity in human acute myeloid leukaemia. Mol Biol Rep. https://doi.org/10.1007/s11033-022-07977-y

    Article  PubMed  PubMed Central  Google Scholar 

  26. Ali KM et al Clinical outcomes and phylogenetic analysis in reflection with three predominant clades of SARS-CoV-2 variants. 0–2 https://doi.org/10.1111/eci.14004

  27. ProtParam tool. https://web.expasy.org/protparam/

  28. Yunus NM et al (2015) Characterisation and clinical significance of FLT3-ITD and non-ITD in Acute myeloid leukaemia patients in Kelantan. Northeast Peninsular Malaysia. https://doi.org/10.7314/APJCP.2015.16.12.4869

    Article  Google Scholar 

  29. Laskowski RA, Thornton JM, VarSite (2020) Disease variants and protein structure. 111–119. https://doi.org/10.1002/pro.3746

  30. Falini B et al (2006) Immunohistochemistry predicts nucleophosmin (NPM) mutations in acute myeloid leukemia. Blood 108:1999–2005

    Article  CAS  PubMed  Google Scholar 

  31. Othman GO, Mohammad NS, Saeed CH (2021) Molecular study of nucleophosmin 1 (NPM1) gene in acute myeloid leukemia in kurdish population. 21:687–692

  32. Yanada M et al (2016) TP53 mutations in older adults with acute myeloid leukemia. Int J Hematol 103:429–435

    Article  CAS  PubMed  Google Scholar 

  33. Dutta S et al (2020) Functional classification of tp53 mutations in acute myeloid leukemia. Cancers (Basel) 12:1–16

    Article  Google Scholar 

  34. Article O (2020) Impact of the variant allele frequency of ASXL1, DNMT3A, JAK2, TET2, TP53, and NPM1 on the Outcomes of patients with newly diagnosed Acute myeloid leukemia. 765–774. https://doi.org/10.1002/cncr.32566

  35. Hamed HR, Al-jumaily RMK, Kadhom AE (2021) Characterization of NPM1 and FLT3-ITD mutations in iraqi patients with AML. 21:133–139

  36. Kiyoi BH et al (1999) Prognostic implication of FLT3 and N- RAS gene mutations in Acute myeloid leukemia. 93:3074–3080

  37. Sazawal S et al (2017) NPM1 and FLT3 mutations in acute myeloid leukemia with normal karyotype. Indian perspective. https://doi.org/10.4103/IJPM.IJPM

    Article  Google Scholar 

  38. Kottaridis PD et al (2001) The presence of a FLT3 internal tandem duplication in patients with acute myeloid leukemia (AML) adds important prognostic information to cytogenetic risk group and response to the first cycle of chemotherapy: analysis of 854 patients from the United King. Blood 98:1752–1759

    Article  CAS  PubMed  Google Scholar 

  39. Rashid PMA, Salih GF (2022) Molecular and computational analysis of spike protein of newly emerged omicron variant in comparison to the delta variant of SARS – CoV – 2 in Iraq. Mol Biol Rep. https://doi.org/10.1007/s11033-022-07545-4

    Article  PubMed  PubMed Central  Google Scholar 

  40. Cheng J, Qu L, Wang J, Cheng L, Wang Y (2018) High expression of FLT3 is a risk factor in leukemia. Mol Med Rep 17:2885–2892

    CAS  PubMed  Google Scholar 

  41. Naseem S et al (2021) NPM1 and FLT3-ITD / TKD gene mutations in. 15:15–26

  42. Hindley A, Catherwood MA, McMullin MF, Mills K (2021) I. significance of npm1 gene mutations in aml. Int J Mol Sci 22,

  43. Sasaki K et al (2020) Impact of the variant allele frequency of ASXL1, DNMT3A, JAK2, TET2, TP53, and NPM1 on the outcomes of patients with newly diagnosed acute myeloid leukemia. Cancer 126:765–774

    Article  CAS  PubMed  Google Scholar 

  44. Döhner H et al (2017) Diagnosis and management of AML in adults: 2017 ELN recommendations from an international expert panel. Blood 129:424–447

    Article  PubMed  PubMed Central  Google Scholar 

  45. Gary Gilliland D, Griffin JD (2002) The roles of FLT3 in hematopoiesis and leukemia. Blood 100:1532–1542

    Article  PubMed  Google Scholar 

  46. Hou HA et al (2012) DNMT3A mutations in acute myeloid leukemia: Stability during disease evolution and clinical implications. Blood 119:559–568

    Article  CAS  PubMed  Google Scholar 

  47. Naoe T, Kiyoi H (2013) Gene mutations of acute myeloid leukemia in the genome era. 165–174. https://doi.org/10.1007/s12185-013-1257-4

  48. Elrhman HAEA, El-Meligui YM, Elalawi SM (2021) Prognostic impact of concurrent DNMT3A, FLT3 and NPM1 gene mutations in Acute myeloid leukemia patients. Clin Lymphoma Myeloma Leuk 21:e960–e969

    Article  CAS  PubMed  Google Scholar 

  49. Easwar A, Siddon AJ (2021) Genetic landscape of myeloproliferative neoplasms with an emphasis on molecular diagnostic laboratory testing. Life 11:1–22

    Article  Google Scholar 

  50. Bumbea H et al (2022) Platelet Defects in Acute Myeloid Leukemia — Potential for Hemorrhagic Events.

  51. Bond J et al (2019) T-cell acute lymphoblastic leukemia 104:1617–1625

    CAS  Google Scholar 

  52. Stone RM et al (2017) Midostaurin plus chemotherapy for Acute myeloid leukemia with a FLT3 mutation. N Engl J Med 377:454–464

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Thiede C et al (2020) Impact of NPM1 / FLT3 -ITD genotypes de fi ned by the 2017 european LeukemiaNet in patients with acute myeloid leukemia. 135:371–380

  54. Barranco R, Fossati F, Candosin S, Orcioni GF, Ventura F (2016) Unexpected sudden death due to acute myeloid leukemia subtype M5: a case report and review of the literature. 207–211. https://doi.org/10.4323/rjlm.2016.207

  55. Kiani BH et al (2020) Artemisinin and its derivatives: a promising cancer therapy. Mol Biol Rep 47:6321–6336

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Chemistry, College of Science, University of Garmian, Kalar, Iraq

    Ayad M. Ali

  2. Department of Biology, College of Science, University of Sulaimani, Sulaymaniyah, Iraq

    Gaza F. Salih

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Contributions

AMA and GFA were involved in the conception of the study idea and the experimental design. AMA conducted the laboratory work, performed data analysis, and prepared the manuscript. GFA provided supervision and guidance throughout the process. Both authors participated in discussions regarding the results and contributed to the finalization of the manuscript.

Corresponding author

Correspondence to Ayad M. Ali.

Ethics declarations

Ethics approval and consent to participate

The ethical aspects of this study were given utmost importance and followed the established guidelines and regulations. Prior to enrollment, all patients provided verbal consent for the collection of blood samples. The study protocol was reviewed and approved by the Ministry of Health’s health directorate, with the approval obtained on August 23, 2021 (protocol NO. 9811). The research investigations were conducted at the University of Sulaimani, College of Science, in accordance with the applicable rules and regulations. The university granted approval for the study (No. 1917/259 on 01/08/2021), ensuring that ethical standards were upheld and necessary protocols were followed throughout the study. The study was conducted in compliance with the research ethical guidelines set by the Ministry of Health. The ethical standards outlined in the Declaration of Helsinki, which governs medical research involving human subjects, were strictly followed. Participants, including the parents of patients under the age of 18, provided both verbal and written consent after being provided with the necessary information and having the opportunity to review the Declaration of Helsinki.

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Competing interests

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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The authors declare no conflict of interest.

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Ali, A.M., Salih, G.F. Molecular and clinical significance of FLT3, NPM1, DNMT3A and TP53 mutations in acute myeloid leukemia patients. Mol Biol Rep 50, 8035–8048 (2023). https://doi.org/10.1007/s11033-023-08680-2

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