Worrillow, L, Baskaran, P, Care, MA et al. (9 more authors) (2016) An ultra-deep sequencing strategy to detect sub-clonal TP53 mutations in presentation chronic lymphocytic leukemia cases using multiple polymerases. Oncogene, 35 (40). pp. 5328-5336. ISSN 0950-9232
Abstract
Chronic lymphocytic leukaemia (CLL) is the most common clonal B-cell disorder characterized by clonal diversity, a relapsing and remitting course, and in its aggressive forms remains largely incurable. Current front-line regimes include agents such as fludarabine, which act primarily via the DNA damage response pathway. Key to this is the transcription factor p53. Mutations in the TP53 gene, altering p53 functionality, are associated with genetic instability, and are present in aggressive CLL. Furthermore, the emergence of clonal TP53 mutations in relapsed CLL, refractory to DNA-damaging therapy, suggests that accurate detection of sub-clonal TP53 mutations prior to and during treatment may be indicative of early relapse. In this study, we describe a novel deep sequencing workflow using multiple polymerases to generate sequencing libraries (MuPol-Seq), facilitating accurate detection of TP53 mutations at a frequency as low as 0.3%, in presentation CLL cases tested. As these mutations were mostly clustered within the regions of TP53 encoding DNA-binding domains, essential for DNA contact and structural architecture, they are likely to be of prognostic relevance in disease progression. The workflow described here has the potential to be implemented routinely to identify rare mutations across a range of diseases.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2016 Macmillan Publishers Limited. This is an author produced version of a paper published in Oncogene. Uploaded in accordance with the publisher's self-archiving policy. |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Medicine and Health (Leeds) > Institute of Molecular Medicine (LIMM) (Leeds) > Section of Experimental Haematology (Leeds) |
Funding Information: | Funder Grant number Kay Kendall Leukaemia Fund KKL693 |
Depositing User: | Symplectic Publications |
Date Deposited: | 15 Mar 2016 12:16 |
Last Modified: | 16 Nov 2016 17:12 |
Published Version: | http://dx.doi.org/10.1038/onc.2016.73 |
Status: | Published |
Publisher: | Nature Publishing Group |
Identification Number: | 10.1038/onc.2016.73 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:96412 |