TY - JOUR
T1 - Genetic screens to identify pathogenic gene variants in the common cancer predisposition Lynch syndrome
AU - Drost, Mark
AU - Lützen, Anne
AU - van Hees, Sandrine
AU - Ferreira, Daniel
AU - Calléja, Fabienne
AU - Zonneveld, José B M
AU - Nielsen, Finn Cilius
AU - Rasmussen, Lene Juel
AU - de Wind, Niels
PY - 2013/6/4
Y1 - 2013/6/4
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.
KW - Amino Acid Substitution
KW - Base Sequence
KW - Colorectal Neoplasms, Hereditary Nonpolyposis
KW - Genetic Predisposition to Disease
KW - Genetic Testing
KW - Genetic Variation
KW - Humans
KW - Microsatellite Repeats
KW - Models, Molecular
KW - Molecular Sequence Data
KW - MutS Homolog 2 Protein
KW - Mutagenesis
KW - Mutation
KW - Reverse Genetics
KW - Sequence Analysis, DNA
KW - Thioguanine
KW - Virulence
U2 - 10.1073/pnas.1220537110
DO - 10.1073/pnas.1220537110
M3 - Journal article
C2 - 23690608
SN - 0027-8424
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
IS - 23
ER -