Pyrosequencing-based methods reveal marked inter-individual differences in oncogene mutation burden in human colorectal tumours

S. Weidlich, K. Walsh, D. Crowther, M. E. Burczynski, G. Feuerstein, Francis Carey, R. J. C. Steele, C. R. Wolf, G. Miele (Lead / Corresponding author), G. Smith (Lead / Corresponding author)

    Research output: Contribution to journalArticle

    20 Citations (Scopus)

    Abstract

    BACKGROUND: The epidermal growth factor receptor-targeted monoclonal antibody cetuximab (Erbitux) was recently introduced for the treatment of metastatic colorectal cancer. Treatment response is dependent on Kirsten-Ras (K-Ras) mutation status, in which the majority of patients with tumour-specific K-Ras mutations fail to respond to treatment. Mutations in the oncogenes B-Raf and PIK3CA (phosphoinositide-3-kinase) may also influence cetuximab response, highlighting the need for a sensitive, accurate and quantitative assessment of tumour mutation burden.

    METHODS: Mutations in K-Ras, B-Raf and PIK3CA were identified by both dideoxy and quantitative pyrosequencing-based methods in a cohort of unselected colorectal tumours (n = 102), and pyrosequencing-based mutation calls correlated with various clinicopathological parameters.

    RESULTS: The use of quantitative pyrosequencing-based methods allowed us to report a 13.7% increase in mutation burden, and to identify low-frequency (<30% mutation burden) mutations not routinely detected by dideoxy sequencing. K-Ras and B-Raf mutations were mutually exclusive and independently associated with a more advanced tumour phenotype.

    CONCLUSION: Pyrosequencing-based methods facilitate the identification of low-frequency tumour mutations and allow more accurate assessment of tumour mutation burden. Quantitative assessment of mutation burden may permit a more detailed evaluation of the role of specific tumour mutations in the pathogenesis and progression of colorectal cancer and may improve future patient selection for targeted drug therapies. British Journal of Cancer (2011) 105, 246-254. doi: 10.1038/bjc.2011.197 www.bjcancer.com Published online 28 June 2011 (C) 2011 Cancer Research UK

    Original languageEnglish
    Pages (from-to)246-254
    Number of pages9
    JournalBritish Journal of Cancer
    Volume105
    Issue number2
    DOIs
    Publication statusPublished - 12 Jul 2011

    Keywords

    • K-Ras
    • Mutation
    • Dideoxy sequencing
    • Pyrosequencing
    • Colorectal tumour
    • Personalised medicine
    • Growth factor receptor
    • Kirsten Ras mutations
    • PIK3CA mutations
    • Prognostic significance
    • Gene mutation
    • Colon cancer
    • CETUXIMAB
    • BRAF

    Cite this

    @article{c0e6086c4482473384ef1c606196fd02,
    title = "Pyrosequencing-based methods reveal marked inter-individual differences in oncogene mutation burden in human colorectal tumours",
    abstract = "BACKGROUND: The epidermal growth factor receptor-targeted monoclonal antibody cetuximab (Erbitux) was recently introduced for the treatment of metastatic colorectal cancer. Treatment response is dependent on Kirsten-Ras (K-Ras) mutation status, in which the majority of patients with tumour-specific K-Ras mutations fail to respond to treatment. Mutations in the oncogenes B-Raf and PIK3CA (phosphoinositide-3-kinase) may also influence cetuximab response, highlighting the need for a sensitive, accurate and quantitative assessment of tumour mutation burden.METHODS: Mutations in K-Ras, B-Raf and PIK3CA were identified by both dideoxy and quantitative pyrosequencing-based methods in a cohort of unselected colorectal tumours (n = 102), and pyrosequencing-based mutation calls correlated with various clinicopathological parameters.RESULTS: The use of quantitative pyrosequencing-based methods allowed us to report a 13.7{\%} increase in mutation burden, and to identify low-frequency (<30{\%} mutation burden) mutations not routinely detected by dideoxy sequencing. K-Ras and B-Raf mutations were mutually exclusive and independently associated with a more advanced tumour phenotype.CONCLUSION: Pyrosequencing-based methods facilitate the identification of low-frequency tumour mutations and allow more accurate assessment of tumour mutation burden. Quantitative assessment of mutation burden may permit a more detailed evaluation of the role of specific tumour mutations in the pathogenesis and progression of colorectal cancer and may improve future patient selection for targeted drug therapies. British Journal of Cancer (2011) 105, 246-254. doi: 10.1038/bjc.2011.197 www.bjcancer.com Published online 28 June 2011 (C) 2011 Cancer Research UK",
    keywords = "K-Ras, Mutation, Dideoxy sequencing, Pyrosequencing, Colorectal tumour, Personalised medicine, Growth factor receptor, Kirsten Ras mutations, PIK3CA mutations, Prognostic significance, Gene mutation, Colon cancer, CETUXIMAB, BRAF",
    author = "S. Weidlich and K. Walsh and D. Crowther and Burczynski, {M. E.} and G. Feuerstein and Francis Carey and Steele, {R. J. C.} and Wolf, {C. R.} and G. Miele and G. Smith",
    year = "2011",
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    language = "English",
    volume = "105",
    pages = "246--254",
    journal = "British Journal of Cancer",
    issn = "0007-0920",
    publisher = "Cancer Research UK",
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    }

    Pyrosequencing-based methods reveal marked inter-individual differences in oncogene mutation burden in human colorectal tumours. / Weidlich, S.; Walsh, K.; Crowther, D.; Burczynski, M. E.; Feuerstein, G.; Carey, Francis; Steele, R. J. C.; Wolf, C. R.; Miele, G. (Lead / Corresponding author); Smith, G. (Lead / Corresponding author).

    In: British Journal of Cancer, Vol. 105, No. 2, 12.07.2011, p. 246-254.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Pyrosequencing-based methods reveal marked inter-individual differences in oncogene mutation burden in human colorectal tumours

    AU - Weidlich, S.

    AU - Walsh, K.

    AU - Crowther, D.

    AU - Burczynski, M. E.

    AU - Feuerstein, G.

    AU - Carey, Francis

    AU - Steele, R. J. C.

    AU - Wolf, C. R.

    AU - Miele, G.

    AU - Smith, G.

    PY - 2011/7/12

    Y1 - 2011/7/12

    N2 - BACKGROUND: The epidermal growth factor receptor-targeted monoclonal antibody cetuximab (Erbitux) was recently introduced for the treatment of metastatic colorectal cancer. Treatment response is dependent on Kirsten-Ras (K-Ras) mutation status, in which the majority of patients with tumour-specific K-Ras mutations fail to respond to treatment. Mutations in the oncogenes B-Raf and PIK3CA (phosphoinositide-3-kinase) may also influence cetuximab response, highlighting the need for a sensitive, accurate and quantitative assessment of tumour mutation burden.METHODS: Mutations in K-Ras, B-Raf and PIK3CA were identified by both dideoxy and quantitative pyrosequencing-based methods in a cohort of unselected colorectal tumours (n = 102), and pyrosequencing-based mutation calls correlated with various clinicopathological parameters.RESULTS: The use of quantitative pyrosequencing-based methods allowed us to report a 13.7% increase in mutation burden, and to identify low-frequency (<30% mutation burden) mutations not routinely detected by dideoxy sequencing. K-Ras and B-Raf mutations were mutually exclusive and independently associated with a more advanced tumour phenotype.CONCLUSION: Pyrosequencing-based methods facilitate the identification of low-frequency tumour mutations and allow more accurate assessment of tumour mutation burden. Quantitative assessment of mutation burden may permit a more detailed evaluation of the role of specific tumour mutations in the pathogenesis and progression of colorectal cancer and may improve future patient selection for targeted drug therapies. British Journal of Cancer (2011) 105, 246-254. doi: 10.1038/bjc.2011.197 www.bjcancer.com Published online 28 June 2011 (C) 2011 Cancer Research UK

    AB - BACKGROUND: The epidermal growth factor receptor-targeted monoclonal antibody cetuximab (Erbitux) was recently introduced for the treatment of metastatic colorectal cancer. Treatment response is dependent on Kirsten-Ras (K-Ras) mutation status, in which the majority of patients with tumour-specific K-Ras mutations fail to respond to treatment. Mutations in the oncogenes B-Raf and PIK3CA (phosphoinositide-3-kinase) may also influence cetuximab response, highlighting the need for a sensitive, accurate and quantitative assessment of tumour mutation burden.METHODS: Mutations in K-Ras, B-Raf and PIK3CA were identified by both dideoxy and quantitative pyrosequencing-based methods in a cohort of unselected colorectal tumours (n = 102), and pyrosequencing-based mutation calls correlated with various clinicopathological parameters.RESULTS: The use of quantitative pyrosequencing-based methods allowed us to report a 13.7% increase in mutation burden, and to identify low-frequency (<30% mutation burden) mutations not routinely detected by dideoxy sequencing. K-Ras and B-Raf mutations were mutually exclusive and independently associated with a more advanced tumour phenotype.CONCLUSION: Pyrosequencing-based methods facilitate the identification of low-frequency tumour mutations and allow more accurate assessment of tumour mutation burden. Quantitative assessment of mutation burden may permit a more detailed evaluation of the role of specific tumour mutations in the pathogenesis and progression of colorectal cancer and may improve future patient selection for targeted drug therapies. British Journal of Cancer (2011) 105, 246-254. doi: 10.1038/bjc.2011.197 www.bjcancer.com Published online 28 June 2011 (C) 2011 Cancer Research UK

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    KW - Mutation

    KW - Dideoxy sequencing

    KW - Pyrosequencing

    KW - Colorectal tumour

    KW - Personalised medicine

    KW - Growth factor receptor

    KW - Kirsten Ras mutations

    KW - PIK3CA mutations

    KW - Prognostic significance

    KW - Gene mutation

    KW - Colon cancer

    KW - CETUXIMAB

    KW - BRAF

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    U2 - 10.1038/bjc.2011.197

    DO - 10.1038/bjc.2011.197

    M3 - Article

    VL - 105

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    EP - 254

    JO - British Journal of Cancer

    JF - British Journal of Cancer

    SN - 0007-0920

    IS - 2

    ER -