Targeted re-sequencing analysis of 25 genes commonly mutated in myeloid disorders in del(5q) myelodysplastic syndromes
Marta Fernandez-Mercado,
Adam Burns,
Andrea Pellagatti,
Aristoteles Giagounidis,
Ulrich Germing,
Xabier Agirre,
Felipe Prosper,
Carlo Aul,
Sally Killick,
James S. Wainscoat,
Anna Schuh,
Jacqueline Boultwood
Affiliations
Marta Fernandez-Mercado
LLR Molecular Haematology Unit, NDCLS, RDM, John Radcliffe Hospital, Oxford, UK
Adam Burns
NIHR Biomedical Research Centre, Oxford, UK
Andrea Pellagatti
LLR Molecular Haematology Unit, NDCLS, RDM, John Radcliffe Hospital, Oxford, UK
Aristoteles Giagounidis
Medizinische Klinik II, St Johannes Hospital, Duisburg, Germany
Ulrich Germing
Department of Hematology, Oncology and Clinical Immunology, Heinrich-Heine-Universität, Düsseldorf, Germany
Xabier Agirre
Division of Cancer and Area of Cell Therapy and Haematology Service, Foundation for Applied Medical Research, Clínica Universitaria, Universidad de Navarra, Pamplona, Spain
Felipe Prosper
Division of Cancer and Area of Cell Therapy and Haematology Service, Foundation for Applied Medical Research, Clínica Universitaria, Universidad de Navarra, Pamplona, Spain
Carlo Aul
Medizinische Klinik II, St Johannes Hospital, Duisburg, Germany
Sally Killick
Department of Haematology, Royal Bournemouth Hospital, Bournemouth, UK
James S. Wainscoat
LLR Molecular Haematology Unit, NDCLS, RDM, John Radcliffe Hospital, Oxford, UK
Anna Schuh
NIHR Biomedical Research Centre, Oxford, UK
Jacqueline Boultwood
LLR Molecular Haematology Unit, NDCLS, RDM, John Radcliffe Hospital, Oxford, UK
Interstitial deletion of chromosome 5q is the most common chromosomal abnormality in myelodysplastic syndromes. The catalogue of genes involved in the molecular pathogenesis of myelodysplastic syndromes is rapidly expanding and next-generation sequencing technology allows detection of these mutations at great depth. Here we describe the design, validation and application of a targeted next-generation sequencing approach to simultaneously screen 25 genes mutated in myeloid malignancies. We used this method alongside single nucleotide polymorphism-array technology to characterize the mutational and cytogenetic profile of 43 cases of early or advanced del(5q) myelodysplastic syndromes. A total of 29 mutations were detected in our cohort. Overall, 45% of early and 66.7% of advanced cases had at least one mutation. Genes with the highest mutation frequency among advanced cases were TP53 and ASXL1 (25% of patients each). These showed a lower mutation frequency in cases of 5q- syndrome (4.5% and 13.6%, respectively), suggesting a role in disease progression in del(5q) myelodysplastic syndromes. Fifty-two percent of mutations identified were in genes involved in epigenetic regulation (ASXL1, TET2, DNMT3A and JAK2). Six mutations had allele frequencies
