Continuous and Periodic Expansion of CAG Repeats in Huntington's Disease R6/1 Mice

Huntington's disease (HD) is one of several neurodegenerative disorders caused by expansion of CAG repeats in a coding gene. Somatic CAG expansion rates in HD vary between organs, and the greatest instability is observed in the brain, correlating with neuropathology. The fundamental mechanisms of somatic CAG repeat instability are poorly understood, but locally formed secondary DNA structures generated during replication and/or repair are believed to underlie triplet repeat expansion. Recent studies in HD mice have demonstrated that mismatch repair (MMR) and base excision repair (BER) proteins are expansion inducing components in brain tissues. This study was designed to simultaneously investigate the rates and modes of expansion in different tissues of HD R6/1 mice in order to further understand the expansion mechanisms in vivo. We demonstrate continuous small expansions in most somatic tissues (exemplified by tail), which bear the signature of many short, probably single-repeat expansions and contractions occurring over time. In contrast, striatum and cortex display a dramatic—and apparently irreversible—periodic expansion. Expansion profiles displaying this kind of periodicity in the expansion process have not previously been reported. These in vivo findings imply that mechanistically distinct expansion processes occur in different tissues.Author Summary: Huntington's disease (HD) is a genetically determined neurodegenerative disorder identified by the presence of a mutation for a long series of CAG repeats (>36 repeats) in the Huntingtin (HTT) gene. Longer repeat sequences cause disease onset at a younger age. The mutation encodes an expanded glutamine tract within the huntingtin protein. This enlarged polyglutamine fragment in the protein leads to the formation of the huntingtin aggregates that are observed in HD brains. The stretch of CAG repeats expands with age in affected brain areas, increasing the length of the polyglutamine tract, and is believed to amplify the effect of the disease. Several HD mouse models display phenotypes relevant to the human disease. We have investigated the rate and modes of expansion in striatum, cortex, and tail in transgenic R6/1 mice. Tail was included as a stable tissue, however we observed a small continuous expansion of CAG repeats in tail tissues. In brain tissues, we identified a periodic expansion process consisting of predominantly seven repeat steps. Our findings point towards a very controlled molecular mechanism as the cause of expansion in the most severely affected tissues, which may provide useful targets that can be used to inhibit disease development.