Differential genomic destabilisation in human cells with pathogenic MSH2 mutations introduced by genome editing

Genki Hayashida, Seijiro Shioi, Kyoko Hidaka, Ryosuke Fujikane, Masumi Hidaka, Toshiki Tsurimoto, Teruhisa Tsuzuki, Shinya Oda, Yoshimichi Nakatsu

Research output: Contribution to journalArticle

Abstract

Repeat destabilisation is variously associated with human disease. In neoplastic diseases, microsatellite instability (MSI) has been regarded as simply reflecting DNA mismatch repair (MMR) deficiency. However, several discrepancies have been pointed out. Firstly, the MSI + phenotype is not uniform in human neoplasms. Established classification utilises the frequency of microsatellite changes, i.e. MSI-H (high) and -L (low), the former regarded as an authentic MMR-defective phenotype. In addition, we have observed the qualitatively distinct modes of MSI, i.e. Type A and Type B. One discrepancy we previously pointed out is that tumours occurring in MMR gene knockout mice exhibited not drastic microsatellite changes typical in MSI-H tumours (i.e. Type B mode) but minor and more subtle alterations (i.e. Type A mode). In the present study, MSH2 mutations reported in Lynch syndrome (LS) kindred have been introduced into HeLa cells using the CRISPR/Cas9 system. The established mutant clones clearly exhibited MMR-defective phenotypes with alkylating agent-tolerance and elevated mutation frequencies. Nevertheless, microsatellites were not markedly destabilised as in MSI-H tumours occurring in LS patients, and all the observed alterations were uniformly Type A, which confirms the results in mice. Our findings suggest added complexities to the molecular mechanisms underlying repeat destabilisation in human genome.

Original languageEnglish
Pages (from-to)24-35
Number of pages12
JournalExperimental Cell Research
Volume377
Issue number1-2
DOIs
Publication statusPublished - Apr 15 2019

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Microsatellite Instability
DNA Mismatch Repair
Mutation
Microsatellite Repeats
Hereditary Nonpolyposis Colorectal Neoplasms
Phenotype
Neoplasms
Clustered Regularly Interspaced Short Palindromic Repeats
DNA Repair-Deficiency Disorders
Gene Knockout Techniques
Alkylating Agents
Human Genome
Mutation Rate
HeLa Cells
Knockout Mice
Gene Editing
Clone Cells

All Science Journal Classification (ASJC) codes

  • Cell Biology

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Differential genomic destabilisation in human cells with pathogenic MSH2 mutations introduced by genome editing. / Hayashida, Genki; Shioi, Seijiro; Hidaka, Kyoko; Fujikane, Ryosuke; Hidaka, Masumi; Tsurimoto, Toshiki; Tsuzuki, Teruhisa; Oda, Shinya; Nakatsu, Yoshimichi.

In: Experimental Cell Research, Vol. 377, No. 1-2, 15.04.2019, p. 24-35.

Research output: Contribution to journalArticle

Hayashida, Genki ; Shioi, Seijiro ; Hidaka, Kyoko ; Fujikane, Ryosuke ; Hidaka, Masumi ; Tsurimoto, Toshiki ; Tsuzuki, Teruhisa ; Oda, Shinya ; Nakatsu, Yoshimichi. / Differential genomic destabilisation in human cells with pathogenic MSH2 mutations introduced by genome editing. In: Experimental Cell Research. 2019 ; Vol. 377, No. 1-2. pp. 24-35.
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