Publication details
Targeted next-generation sequencing in chronic lymphocytic leukemia: a high-throughput yet tailored approach will facilitate implementation within a clinical setting
| Authors | |
|---|---|
| Year of publication | 2015 |
| Type | Article in Periodical |
| Magazine / Source | Haematologica |
| MU Faculty or unit | |
| Citation | SUTTON, Lesley-Ann, Viktor LJUNGSTRÖM, Larry MANSOURI, Emma YOUNG, Diego CORTESE, Veronika NAVRKALOVÁ, Jitka MALČÍKOVÁ, Alice F. MUGGEN, Martin TRBUŠEK, Panagiotis PANAGIOTIDIS, Frederic DAVI, Chrysoula BELESSI, Anton W. LANGERAK, Paolo GHIA, Šárka POSPÍŠILOVÁ, Kostas STAMATOPOULOS and Richard ROSENQUIST. Targeted next-generation sequencing in chronic lymphocytic leukemia: a high-throughput yet tailored approach will facilitate implementation within a clinical setting. Haematologica. Pavia (Italy): Ferrata Storti Foundation, 2015, vol. 100, No 3, p. 370-376. ISSN 0390-6078. Available from: https://dx.doi.org/10.3324/haematol.2014.109777. |
| web | http://www.haematologica.org/content/100/3/370.full-text.pdf+html |
| Doi | http://dx.doi.org/10.3324/haematol.2014.109777 |
| Field | Oncology and hematology |
| Keywords | Chronic Lymphocytic Leukemia; Cytogenetics and Molecular Genetics; Next-generation sequencing; Prognosis; Targeted resequencing |
| Attached files | |
| Description | Next-generation sequencing has revealed novel recurrent mutations in chronic lymphocytic leukemia, particularly in patients with aggressive disease. Here, we explored targeted re-sequencing as a novel strategy to assess the mutation status of genes with prognostic potential. To this end, we utilized the HaloPlex targeted enrichment technology and designed a panel including nine genes: ATM, BIRC3, MYD88, NOTCH1, SF3B1 and TP53, that have been linked to chronic lymphocytic leukemia prognosis, and KLHL6, POT1 and XPO1, that are less characterized but were found recurrently mutated in various sequencing studies. A total of 188 chronic lymphocytic leukemia patients with poor-prognostic features (unmutated IGHV, n=137; IGHV3-21 subset #2, n=51) were sequenced on the HiSeq 2000 and data were analyzed using well-established bioinformatics tools. Using a conservative cutoff of 10% for the mutant allele, we found that 114/180 (63%) patients carried at least one mutation, with mutations in ATM, BIRC3, NOTCH1, SF3B1 and TP53 accounting for 149/177 (84%) of all mutations. We selected 155 mutations for Sanger validation (variant allele frequency, 10-99%) and 93% (144/155) of mutations were confirmed; notably, all 11 discordant variants had a variant allele frequency between 11-27%, hence at the detection limit of conventional Sanger sequencing. Technical precision was assessed by repeating the entire Haloplex procedure for 63 patients with 77/82 (94%) mutations demonstrating concordance. In summary, this study demonstrates targeted next-generation sequencing as an accurate and reproducible technique potentially suitable for routine screening, eventually as a stand-alone test without the need for confirmation by Sanger sequencing. |
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