Whole exome sequencing of an asbestos-induced wild-type murine model of malignant mesothelioma

Sophie Sneddon, Ann Marie Patch, Ian M. Dick, Stephen H. Kazakoff, John V. Pearson, Nicola Waddell, Richard J.N. Allcock, Robert A. Holt, Bruce W.S. Robinson, Jenette Creaney

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    Abstract

    Background: Malignant mesothelioma (MM) is an aggressive cancer of the pleural and peritoneal cavities caused by exposure to asbestos. Asbestos-induced mesotheliomas in wild-type mice have been used extensively as a preclinical model because they are phenotypically identical to their human counterpart. However, it is not known if the genetic lesions in these mice tumours are similar to in the human disease, a prerequisite for any new preclinical studies that target genetic abnormalities. Methods: We performed whole exome sequencing of fifteen asbestos-induced murine MM tumour cell lines from BALB/c, CBA and C57BL/6 mouse strains and compared the somatic mutations and copy number variations with those recurrently reported in human MM. We then catalogued and characterised the mutational landscape of the wild-type murine MM tumours. Quantitative RT-PCR was used to interrogate the expression of key MM genes of interest in the mRNA. Results: Consistent with human MM tumours, we identified homozygous loss of the tumour suppressor Cdkn2a in 14/15 tumours. One tumour retained the first exon of both of the p16INK4a and p19ARF isoforms though this tumour also contained genetic amplification of Myc resulting in increased expression of the c-Myc proto-oncogene in the mRNA. There were no chromosomal losses in either the Bap1 or Nf2 regions. One tumour harbored homozygous loss of Trp53 in the DNA. Mutation rates were similar in tumours generated in the CBA and C57BL/6 strains when compared to human MM. Interestingly, all BALB/c tumour lines displayed high mutational loads, consistent with the known mutator phenotype of the host strain. The Wnt, MAPK and Jak-STAT signaling pathways were found to be the most commonly affected biological pathways. Mutations and copy number deletions also occurred in the Hedgehog and Hippo pathways. Conclusions: These data suggest that in the wild-type murine model asbestos causes mesotheliomas in a similar way to in human MM. This further supports the notion that the murine model of MM represents a genuine homologue of the human disease, something uncommon in cancer, and is thus a valuable tool to provide insight into MM tumour development and to aide the search for novel therapeutic strategies.

    Original languageEnglish
    Article number396
    JournalBMC Cancer
    Volume17
    Issue number1
    DOIs
    Publication statusPublished - 2 Jun 2017

    Fingerprint

    Exome
    Asbestos
    Neoplasms
    Mesothelioma
    Malignant Mesothelioma
    Pleural Cavity
    Messenger RNA
    Mutation
    myc Genes
    Hedgehogs
    Peritoneal Cavity
    Mutation Rate
    Tumor Cell Line
    Inbred C57BL Mouse

    Cite this

    Sneddon, Sophie ; Patch, Ann Marie ; Dick, Ian M. ; Kazakoff, Stephen H. ; Pearson, John V. ; Waddell, Nicola ; Allcock, Richard J.N. ; Holt, Robert A. ; Robinson, Bruce W.S. ; Creaney, Jenette. / Whole exome sequencing of an asbestos-induced wild-type murine model of malignant mesothelioma. In: BMC Cancer. 2017 ; Vol. 17, No. 1.
    @article{5fde74e81f7c46b58d867807de334ea6,
    title = "Whole exome sequencing of an asbestos-induced wild-type murine model of malignant mesothelioma",
    abstract = "Background: Malignant mesothelioma (MM) is an aggressive cancer of the pleural and peritoneal cavities caused by exposure to asbestos. Asbestos-induced mesotheliomas in wild-type mice have been used extensively as a preclinical model because they are phenotypically identical to their human counterpart. However, it is not known if the genetic lesions in these mice tumours are similar to in the human disease, a prerequisite for any new preclinical studies that target genetic abnormalities. Methods: We performed whole exome sequencing of fifteen asbestos-induced murine MM tumour cell lines from BALB/c, CBA and C57BL/6 mouse strains and compared the somatic mutations and copy number variations with those recurrently reported in human MM. We then catalogued and characterised the mutational landscape of the wild-type murine MM tumours. Quantitative RT-PCR was used to interrogate the expression of key MM genes of interest in the mRNA. Results: Consistent with human MM tumours, we identified homozygous loss of the tumour suppressor Cdkn2a in 14/15 tumours. One tumour retained the first exon of both of the p16INK4a and p19ARF isoforms though this tumour also contained genetic amplification of Myc resulting in increased expression of the c-Myc proto-oncogene in the mRNA. There were no chromosomal losses in either the Bap1 or Nf2 regions. One tumour harbored homozygous loss of Trp53 in the DNA. Mutation rates were similar in tumours generated in the CBA and C57BL/6 strains when compared to human MM. Interestingly, all BALB/c tumour lines displayed high mutational loads, consistent with the known mutator phenotype of the host strain. The Wnt, MAPK and Jak-STAT signaling pathways were found to be the most commonly affected biological pathways. Mutations and copy number deletions also occurred in the Hedgehog and Hippo pathways. Conclusions: These data suggest that in the wild-type murine model asbestos causes mesotheliomas in a similar way to in human MM. This further supports the notion that the murine model of MM represents a genuine homologue of the human disease, something uncommon in cancer, and is thus a valuable tool to provide insight into MM tumour development and to aide the search for novel therapeutic strategies.",
    keywords = "Asbestos, Cdkn2a, Exome sequencing, Mesothelioma, Mouse model, Wild-type",
    author = "Sophie Sneddon and Patch, {Ann Marie} and Dick, {Ian M.} and Kazakoff, {Stephen H.} and Pearson, {John V.} and Nicola Waddell and Allcock, {Richard J.N.} and Holt, {Robert A.} and Robinson, {Bruce W.S.} and Jenette Creaney",
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    Whole exome sequencing of an asbestos-induced wild-type murine model of malignant mesothelioma. / Sneddon, Sophie; Patch, Ann Marie; Dick, Ian M.; Kazakoff, Stephen H.; Pearson, John V.; Waddell, Nicola; Allcock, Richard J.N.; Holt, Robert A.; Robinson, Bruce W.S.; Creaney, Jenette.

    In: BMC Cancer, Vol. 17, No. 1, 396, 02.06.2017.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Whole exome sequencing of an asbestos-induced wild-type murine model of malignant mesothelioma

    AU - Sneddon, Sophie

    AU - Patch, Ann Marie

    AU - Dick, Ian M.

    AU - Kazakoff, Stephen H.

    AU - Pearson, John V.

    AU - Waddell, Nicola

    AU - Allcock, Richard J.N.

    AU - Holt, Robert A.

    AU - Robinson, Bruce W.S.

    AU - Creaney, Jenette

    PY - 2017/6/2

    Y1 - 2017/6/2

    N2 - Background: Malignant mesothelioma (MM) is an aggressive cancer of the pleural and peritoneal cavities caused by exposure to asbestos. Asbestos-induced mesotheliomas in wild-type mice have been used extensively as a preclinical model because they are phenotypically identical to their human counterpart. However, it is not known if the genetic lesions in these mice tumours are similar to in the human disease, a prerequisite for any new preclinical studies that target genetic abnormalities. Methods: We performed whole exome sequencing of fifteen asbestos-induced murine MM tumour cell lines from BALB/c, CBA and C57BL/6 mouse strains and compared the somatic mutations and copy number variations with those recurrently reported in human MM. We then catalogued and characterised the mutational landscape of the wild-type murine MM tumours. Quantitative RT-PCR was used to interrogate the expression of key MM genes of interest in the mRNA. Results: Consistent with human MM tumours, we identified homozygous loss of the tumour suppressor Cdkn2a in 14/15 tumours. One tumour retained the first exon of both of the p16INK4a and p19ARF isoforms though this tumour also contained genetic amplification of Myc resulting in increased expression of the c-Myc proto-oncogene in the mRNA. There were no chromosomal losses in either the Bap1 or Nf2 regions. One tumour harbored homozygous loss of Trp53 in the DNA. Mutation rates were similar in tumours generated in the CBA and C57BL/6 strains when compared to human MM. Interestingly, all BALB/c tumour lines displayed high mutational loads, consistent with the known mutator phenotype of the host strain. The Wnt, MAPK and Jak-STAT signaling pathways were found to be the most commonly affected biological pathways. Mutations and copy number deletions also occurred in the Hedgehog and Hippo pathways. Conclusions: These data suggest that in the wild-type murine model asbestos causes mesotheliomas in a similar way to in human MM. This further supports the notion that the murine model of MM represents a genuine homologue of the human disease, something uncommon in cancer, and is thus a valuable tool to provide insight into MM tumour development and to aide the search for novel therapeutic strategies.

    AB - Background: Malignant mesothelioma (MM) is an aggressive cancer of the pleural and peritoneal cavities caused by exposure to asbestos. Asbestos-induced mesotheliomas in wild-type mice have been used extensively as a preclinical model because they are phenotypically identical to their human counterpart. However, it is not known if the genetic lesions in these mice tumours are similar to in the human disease, a prerequisite for any new preclinical studies that target genetic abnormalities. Methods: We performed whole exome sequencing of fifteen asbestos-induced murine MM tumour cell lines from BALB/c, CBA and C57BL/6 mouse strains and compared the somatic mutations and copy number variations with those recurrently reported in human MM. We then catalogued and characterised the mutational landscape of the wild-type murine MM tumours. Quantitative RT-PCR was used to interrogate the expression of key MM genes of interest in the mRNA. Results: Consistent with human MM tumours, we identified homozygous loss of the tumour suppressor Cdkn2a in 14/15 tumours. One tumour retained the first exon of both of the p16INK4a and p19ARF isoforms though this tumour also contained genetic amplification of Myc resulting in increased expression of the c-Myc proto-oncogene in the mRNA. There were no chromosomal losses in either the Bap1 or Nf2 regions. One tumour harbored homozygous loss of Trp53 in the DNA. Mutation rates were similar in tumours generated in the CBA and C57BL/6 strains when compared to human MM. Interestingly, all BALB/c tumour lines displayed high mutational loads, consistent with the known mutator phenotype of the host strain. The Wnt, MAPK and Jak-STAT signaling pathways were found to be the most commonly affected biological pathways. Mutations and copy number deletions also occurred in the Hedgehog and Hippo pathways. Conclusions: These data suggest that in the wild-type murine model asbestos causes mesotheliomas in a similar way to in human MM. This further supports the notion that the murine model of MM represents a genuine homologue of the human disease, something uncommon in cancer, and is thus a valuable tool to provide insight into MM tumour development and to aide the search for novel therapeutic strategies.

    KW - Asbestos

    KW - Cdkn2a

    KW - Exome sequencing

    KW - Mesothelioma

    KW - Mouse model

    KW - Wild-type

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    U2 - 10.1186/s12885-017-3382-6

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    VL - 17

    JO - BMC Cancer

    JF - BMC Cancer

    SN - 1471-2407

    IS - 1

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    ER -