F. Nollet PhD, MSc

Diagnostic testing in myeloid malignancies by next-generation sequencing: recommendations from the Commission Personalised Medicine

SUMMARY

Molecular diagnostics have an increasing impact on diagnosis, risk stratification and targeted treatment in haemato-oncology. In the framework of a pilot study for the implementation of next-generation sequencing in the Belgian healthcare system, the Commission of Personalised Medicine was founded to give professional and evidence-based advice on the molecular analysis in haemato-oncology. This paper describes its recommendations for NGS analysis in myeloid malignancies. In addition, the minimally required set of genes that must be analysed is defined and algorithms for molecular workflow in myeloid malignancies are proposed.

(BELG J HEMATOL 2019;10(6):241–9)

Targeted next generation DNA sequencing for the detection of clonal haematopoiesis in idiopathic cytopaenia of undetermined significance (ICUS)

SUMMARY

Patients lacking diagnostic criteria for myelodysplastic syndrome, but who show an unexplained persistent cytopaenia are classified as patients suffering from idiopathic cytopaenia of undetermined significance (ICUS). A fraction of these patients carry somatic mutations in genes which are also mutated in myeloid neoplasms. The significance of these mutations in ICUS patients is not well known and only few research papers have tried to correlate them with clinical outcome. ICUS patients carrying somatic mutations seem to have a higher progression rate to myeloid malignancies compared to unmutated patients. Some mutation profiles also show lower overall survival, similar to patients with (low-risk) myelodysplastic syndrome. Therefore, it seems useful to screen for somatic mutations in cytopaenic patients. The goal of this paper is to review recent literature regarding the significance of somatic mutations in cytopaenic patients and propose a screening protocol by evaluating a test protocol at the AZ Sint-Jan hospital Brugge-Oostende.

(BELG J HEMATOL 2019;10(6):231–40)

PP48 Validation of first trimester non-invasive fetal RhD genotyping for targeted antenatal anti-D prophylaxis

P07 Refractory mediastinal myeloid sarcoma preceding an acute myeloid leukemia with t(10;11) chromosome translocation

Targeted next generation sequencing in myeloid neoplasms

Summary

For most haematological disease entities whole genome and/or exome sequencing efforts identified a core set of recurrently mutated genes. Multiplex DNA mutation screening proves to be highly applicable for myeloid malignancies, since mutations in many genes, e.g. FLT3, NPM1, CEBPA, KIT, DNMT3A, IDH1, IDH2, TET2, ASXL1, RUNX1, SF3B1, SRSF2, U2AF1, ZRSR2, TP53, STAG2, SMC1A, SMC3, RAD21, PHF6, RAS, EZH2, ETV6, JAK2, MPL, CALR, SETBP1, CSF3R, are described to be significantly associated with diagnosis, disease subtyping, prognostication, and/or for tailoring therapy. Obviously, their analysis is no longer feasible using conventional, single gene molecular diagnostic techniques, urging the use of a multi-gene ‘pan-myeloid’ Next Generation Sequencing panel.

(BELG J HEMATOL 2016; 7(3):98–102)

Challenges and pitfalls of next generation sequencing in molecular haematology

Summary

In the past few years the cost of next generation sequencing decreased substantially and the technology has significantly matured, allowing for its introduction into clinical practice. For most haematological disease entities whole genome and/or exome sequencing efforts identified a core set of recurrently mutated genes. Several of these genes are expected to be included in the upcoming revision of the 2008 edition of the World Health Organisation classification of haematological malignancies. Next generation sequencing technology allows transforming current single gene mutation analysis into multiplexed mutational profiling. Next generation ‘deep’ sequencing is a promising new tool to monitor minimal disease burden and to detect mutations within malignant subclones. With current platforms point mutations can be detected with a sensitivity of 1–5% mutant DNA. For indel mutations or clonal IG/TCR rearrangements sensitivity in the range of 10⁻⁵ can be reached. In this review we will highlight the opportunities and challenges of the introduction of next generation sequencing technology into a setting where it will contribute significantly to individual patient cancer management.

(BELG J HEMATOL 2016;7(2):63–8)

PP4.3 Detection and Monitoring of BCR-ABL1 Kinase Domain Mutations by Next Generation Sequencing