Genetic screens to identify pathogenic gene variants in the common cancer predisposition Lynch syndrome

Anne Lützen, Mark Drost, Sandrine Van Hees, Daniel Ferreira, Fabienne Calleja, Jose Zonneveld, Finn Cilius Nielsen, Lene Juel Rasmussen, Niels de Wind

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    Resumé

    In many individuals suspected of the common cancer predisposition Lynch syndrome, variants of unclear significance (VUS), rather than an obviously pathogenic mutations, are identified in one of the DNA mismatch repair (MMR) genes. The uncertainty of whether such VUS inactivate MMR, and therefore are pathogenic, precludes targeted healthcare for both carriers and their relatives. To facilitate the identification of pathogenic VUS, we have developed an in cellulo genetic screen-based procedure for the large-scale mutagenization, identification, and cataloging of residues of MMR genes critical for MMR gene function. When a residue identified as mutated in an individual suspected of Lynch syndrome is listed as critical in such a reverse diagnosis catalog, there is a high probability that the corresponding human VUS is pathogenic. To investigate the applicability of this approach, we have generated and validated a prototypic reverse diagnosis catalog for the MMR gene MutS Homolog 2 (Msh2) by mutagenizing, identifying, and cataloging 26 deleterious mutations in 23 amino acids. Extensive in vivo and in vitro analysis of mutants listed in the catalog revealed both recessive and dominant-negative phenotypes. Nearly half of these critical residues match with VUS previously identified in individuals suspected of Lynch syndrome. This aids in the assignment of pathogenicity to these human VUS and validates the approach described here as a diagnostic tool. In a wider perspective, this work provides a model for the translation of personalized genomics into targeted healthcare.
    OriginalsprogEngelsk
    TidsskriftProceedings of the National Academy of Sciences of the United States of America
    Vol/bind110
    Udgave nummer23
    Sider (fra-til)9403-9408
    ISSN0027-8424
    DOI
    StatusUdgivet - 2013

    Citer dette

    Lützen, Anne ; Drost, Mark ; Van Hees, Sandrine ; Ferreira, Daniel ; Calleja, Fabienne ; Zonneveld, Jose ; Nielsen, Finn Cilius ; Rasmussen, Lene Juel ; de Wind , Niels. / Genetic screens to identify pathogenic gene variants in the common cancer predisposition Lynch syndrome. I: Proceedings of the National Academy of Sciences of the United States of America. 2013 ; Bind 110, Nr. 23. s. 9403-9408.
    @article{188dd9adfb6f49749268c895dd86f80f,
    title = "Genetic screens to identify pathogenic gene variants in the common cancer predisposition Lynch syndrome",
    abstract = "In many individuals suspected of the common cancer predisposition Lynch syndrome, variants of unclear significance (VUS), rather than an obviously pathogenic mutations, are identified in one of the DNA mismatch repair (MMR) genes. The uncertainty of whether such VUS inactivate MMR, and therefore are pathogenic, precludes targeted healthcare for both carriers and their relatives. To facilitate the identification of pathogenic VUS, we have developed an in cellulo genetic screen-based procedure for the large-scale mutagenization, identification, and cataloging of residues of MMR genes critical for MMR gene function. When a residue identified as mutated in an individual suspected of Lynch syndrome is listed as critical in such a reverse diagnosis catalog, there is a high probability that the corresponding human VUS is pathogenic. To investigate the applicability of this approach, we have generated and validated a prototypic reverse diagnosis catalog for the MMR gene MutS Homolog 2 (Msh2) by mutagenizing, identifying, and cataloging 26 deleterious mutations in 23 amino acids. Extensive in vivo and in vitro analysis of mutants listed in the catalog revealed both recessive and dominant-negative phenotypes. Nearly half of these critical residues match with VUS previously identified in individuals suspected of Lynch syndrome. This aids in the assignment of pathogenicity to these human VUS and validates the approach described here as a diagnostic tool. In a wider perspective, this work provides a model for the translation of personalized genomics into targeted healthcare.",
    author = "Anne L{\"u}tzen and Mark Drost and {Van Hees}, Sandrine and Daniel Ferreira and Fabienne Calleja and Jose Zonneveld and Nielsen, {Finn Cilius} and Rasmussen, {Lene Juel} and {de Wind}, Niels",
    year = "2013",
    doi = "10.1073/pnas.1220537110",
    language = "English",
    volume = "110",
    pages = "9403--9408",
    journal = "Proceedings of the National Academy of Sciences of the United States of America",
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    Lützen, A, Drost, M, Van Hees, S, Ferreira, D, Calleja, F, Zonneveld, J, Nielsen, FC, Rasmussen, LJ & de Wind , N 2013, 'Genetic screens to identify pathogenic gene variants in the common cancer predisposition Lynch syndrome', Proceedings of the National Academy of Sciences of the United States of America, bind 110, nr. 23, s. 9403-9408. https://doi.org/10.1073/pnas.1220537110

    Genetic screens to identify pathogenic gene variants in the common cancer predisposition Lynch syndrome. / Lützen, Anne; Drost, Mark; Van Hees, Sandrine; Ferreira, Daniel; Calleja, Fabienne ; Zonneveld, Jose; Nielsen, Finn Cilius; Rasmussen, Lene Juel; de Wind , Niels.

    I: Proceedings of the National Academy of Sciences of the United States of America, Bind 110, Nr. 23, 2013, s. 9403-9408.

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    TY - JOUR

    T1 - Genetic screens to identify pathogenic gene variants in the common cancer predisposition Lynch syndrome

    AU - Lützen, Anne

    AU - Drost, Mark

    AU - Van Hees, Sandrine

    AU - Ferreira, Daniel

    AU - Calleja, Fabienne

    AU - Zonneveld, Jose

    AU - Nielsen, Finn Cilius

    AU - Rasmussen, Lene Juel

    AU - de Wind , Niels

    PY - 2013

    Y1 - 2013

    N2 - In many individuals suspected of the common cancer predisposition Lynch syndrome, variants of unclear significance (VUS), rather than an obviously pathogenic mutations, are identified in one of the DNA mismatch repair (MMR) genes. The uncertainty of whether such VUS inactivate MMR, and therefore are pathogenic, precludes targeted healthcare for both carriers and their relatives. To facilitate the identification of pathogenic VUS, we have developed an in cellulo genetic screen-based procedure for the large-scale mutagenization, identification, and cataloging of residues of MMR genes critical for MMR gene function. When a residue identified as mutated in an individual suspected of Lynch syndrome is listed as critical in such a reverse diagnosis catalog, there is a high probability that the corresponding human VUS is pathogenic. To investigate the applicability of this approach, we have generated and validated a prototypic reverse diagnosis catalog for the MMR gene MutS Homolog 2 (Msh2) by mutagenizing, identifying, and cataloging 26 deleterious mutations in 23 amino acids. Extensive in vivo and in vitro analysis of mutants listed in the catalog revealed both recessive and dominant-negative phenotypes. Nearly half of these critical residues match with VUS previously identified in individuals suspected of Lynch syndrome. This aids in the assignment of pathogenicity to these human VUS and validates the approach described here as a diagnostic tool. In a wider perspective, this work provides a model for the translation of personalized genomics into targeted healthcare.

    AB - In many individuals suspected of the common cancer predisposition Lynch syndrome, variants of unclear significance (VUS), rather than an obviously pathogenic mutations, are identified in one of the DNA mismatch repair (MMR) genes. The uncertainty of whether such VUS inactivate MMR, and therefore are pathogenic, precludes targeted healthcare for both carriers and their relatives. To facilitate the identification of pathogenic VUS, we have developed an in cellulo genetic screen-based procedure for the large-scale mutagenization, identification, and cataloging of residues of MMR genes critical for MMR gene function. When a residue identified as mutated in an individual suspected of Lynch syndrome is listed as critical in such a reverse diagnosis catalog, there is a high probability that the corresponding human VUS is pathogenic. To investigate the applicability of this approach, we have generated and validated a prototypic reverse diagnosis catalog for the MMR gene MutS Homolog 2 (Msh2) by mutagenizing, identifying, and cataloging 26 deleterious mutations in 23 amino acids. Extensive in vivo and in vitro analysis of mutants listed in the catalog revealed both recessive and dominant-negative phenotypes. Nearly half of these critical residues match with VUS previously identified in individuals suspected of Lynch syndrome. This aids in the assignment of pathogenicity to these human VUS and validates the approach described here as a diagnostic tool. In a wider perspective, this work provides a model for the translation of personalized genomics into targeted healthcare.

    U2 - 10.1073/pnas.1220537110

    DO - 10.1073/pnas.1220537110

    M3 - Journal article

    VL - 110

    SP - 9403

    EP - 9408

    JO - Proceedings of the National Academy of Sciences of the United States of America

    JF - Proceedings of the National Academy of Sciences of the United States of America

    SN - 0027-8424

    IS - 23

    ER -