Use of CRISPR-modified human stem cell organoids to study the origin of mutational signatures in cancer.
American Association for the Advancement of Science (AAAS)
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Drost, J., van Boxtel, R., Blokzijl, F., Mizutani, T., Sasaki, N., Sasselli, V., de Ligt, J., et al. (2017). Use of CRISPR-modified human stem cell organoids to study the origin of mutational signatures in cancer.. Science, 358 (6360), 234-238. https://doi.org/10.1126/science.aao3130
Mutational processes underlie cancer initiation and progression. Signatures of these processes in cancer genomes may explain cancer etiology and could hold diagnostic and prognostic value. We developed a strategy that can be used to explore the origin of cancer-associated mutational signatures. We used CRISPR-Cas9 technology to delete key DNA repair genes in human colon organoids, followed by delayed subcloning and whole-genome sequencing. We found that mutation accumulation in organoids deficient in the mismatch repair gene MLH1 is driven by replication errors and accurately models the mutation profiles observed in mismatch repair-deficient colorectal cancers. Application of this strategy to the cancer predisposition gene NTHL1, which encodes a base excision repair protein, revealed a mutational footprint (signature 30) previously observed in a breast cancer cohort. We show that signature 30 can arise from germline NTHL1 mutations.
Colon, Organoids, Stem Cells, Humans, Neoplasms, Breast Neoplasms, Colorectal Neoplasms, Deoxyribonuclease (Pyrimidine Dimer), DNA Repair, DNA Replication, Mutagenesis, Germ-Line Mutation, Female, DNA Mismatch Repair, INDEL Mutation, CRISPR-Cas Systems, MutL Protein Homolog 1
External DOI: https://doi.org/10.1126/science.aao3130
This record's URL: https://www.repository.cam.ac.uk/handle/1810/283071