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

Research output: Contribution to journalJournal articleResearchpeer-review

Mark Drost, Anne Lützen, Sandrine van Hees, Daniel Ferreira, Fabienne Calléja, José B M Zonneveld, Finn Cilius Nielsen, Lene Juel Rasmussen, Niels de Wind

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.
Original languageEnglish
JournalProceedings of the National Academy of Science of the United States of America
Volume110
Issue number23
Pages (from-to)9403-8
Number of pages6
ISSN0027-8424
DOIs
Publication statusPublished - 4 Jun 2013

ID: 47458484