Replacement of normal with mutant alleles in the genome of normal human cells unveils mutation-specific drug responses.
Mutations in oncogenes and tumor suppressor genes are responsible for tumorigenesis and represent favored therapeutic targets in oncology. We exploited homologous recombination to knock-in individual cancer mutations in the genome of nontransformed human cells. Sequential introduction of multiple mutations was also achieved, demonstrating the potential of this strategy to construct tumor progression models. Knock-in cells displayed allele-specific activation of signaling pathways and mutation-specific phenotypes different from those obtainable by ectopic oncogene expression. Profiling of a library of pharmacological agents on the mutated cells showed striking sensitivity or resistance phenotypes to pathway-targeted drugs, often matching those of tumor cells carrying equivalent cancer mutations. Thus, knock-in of single or multiple cancer alleles provides a pharmacogenomic platform for the rational design of targeted therapies.
Di Nicolantonio
Federica
F
Laboratory of Molecular Genetics, Institute for Cancer Research and Treatment, University of Turin Medical School, I-10060 Candiolo, Turin, Italy.
Arena
Sabrina
S
Gallicchio
Margherita
M
Zecchin
Davide
D
Martini
Miriam
M
Flonta
Simona Emilia
SE
Stella
Giulia Maria
GM
Lamba
Simona
S
Cancelliere
Carlotta
C
Russo
Mariangela
M
Geuna
Massimo
M
Appendino
Giovanni
G
Fantozzi
Roberto
R
Medico
Enzo
E
Bardelli
Alberto
A
D000483:Alleles; D000970:Antineoplastic Agents / Q000494:pharmacology* / Q000627:therapeutic use; D002460:Cell Line; D016503:Drug Delivery Systems / Q000379:methods; D004354:Drug Screening Assays, Antitumor; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D055879:Gene Knock-In Techniques; D016147:Genes, Tumor Suppressor; D006801:Humans; D008954:Models, Biological; D009154:Mutation; D009369:Neoplasms / Q000188:drug therapy* / Q000235:genetics*
D000970:Antineoplastic Agents