Defining the mutational profile of lower-risk myelodysplastic neoplasm patients with respect to disease progression using next-generation sequencing and pyrosequencing

Monika Adamska; Ewelina Kowal-Wiśniewska; Joanna Czerwińska-Rybak; Katarzyna Kiwerska; Marta Barańska; Weronika Gronowska; Jagoda Loba; Katarzyna Brzeźniakiewicz-Janus; Ewa Wasilewska; Aleksandra Łanocha; Małgorzata Jarmuż-Szymczak; Lidia Gil

doi:10.5114/wo.2023.135365

Defining the mutational profile of lower-risk myelodysplastic neoplasm patients with respect to disease progression using next-generation sequencing and pyrosequencing

Contemp Oncol (Pozn). 2023;27(4):269-279. doi: 10.5114/wo.2023.135365. Epub 2024 Feb 17.

Authors

Affiliations

¹ Department of Haematology and Bone Marrow Transplantation, Poznań University of Medical Sciences, Poznań, Poland.
² Doctoral School, Poznań University of Medical Sciences, Poznań, Poland.
³ Institute of Human Genetics, Polish Academy of Sciences, Poznań, Poland.
⁴ Student Scientific Society, Poznań University of Medical Sciences, Poznań, Poland.
⁵ Department of Haematology, Oncology, and Radiotherapy, University of Zielona Góra, Multi-specialist Hospital Gorzów Wielkopolski, Poland.
⁶ Department of Haematology, Medical University of Białystok, Białystok, Poland.
⁷ Department of Haematology with Bone Marrow Transplantation Unit, University Hospital No. 1 of Pomeranian Medical University, Szczecin, Poland.

Abstract

Introduction: Lower-risk myelodysplastic neoplasms (LR-MDS) comprise the majority of MDS. Despite favourable prognoses, some patients remain at risk of rapid progression. We aimed to define the mutational profile of LR-MDS using next-generation sequencing (NGS), Sanger Sequencing (SSeq), and pyrosequencing.

Material and methods: Samples from 5 primary LR-MDS (67 exons of SF3B1, U2AF1, SRSF2, ZRSR2, TET2, ASXL1, DNMT3A, TP53, and RUNX1 genes) were subjected to NGS. Next, a genomic study was performed to test for the presence of identified DNA sequence variants on a larger group of LR-MDS patients (25 bone marrow [BM], 3 saliva [SAL], and one peripheral blood [PB] sample/s). Both SSeq (all selected DNA sequence variants) and pyrosequencing (9 selected DNA sequence variants) were performed.

Results: Next-generation sequencing results identified 13 DNA sequence variants in 7 genes, comprising 8 mutations in 6 genes (ASXL1, DNMT3A, RUNX1, SF3B1, TET2, ZRSR2) in LR-MDS. The presence of 8 DNA variants was detected in the expanded LR-MDS group using SSeq and pyrosequencing. Mutation acquisition was observed during LR-MDS progression. Four LR-MDS and one acute myeloid leukaemia myelodysplasia-related patient exhibited the presence of at least one mutation. ASXL1 and SF3B1 alterations were most commonly observed (2 patients). Five DNA sequence variants detected in BM (patients: 9, 13) were also present in SAL.

Conclusions: We suggest using NGS to determine the LR-MDS mutational profile at diagnosis and suspicion of disease progression. Moreover, PB and SAL molecular testing represent useful tools for monitoring LR-MDS at higher risk of progression. However, the results need to be confirmed in a larger group.

Keywords: NGS; acute myeloid leukaemia myelodysplasia-related; allogenic hematopoietic cell transplantation; lower-risk myelodysplastic neoplasms; pyrosequencing.