Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, United States of America.
PLoS One. 2011;6(8):e23647. doi: 10.1371/journal.pone.0023647. Epub 2011 Aug 29.
Age at onset of Huntington's disease (HD) is largely determined by the CAG trinucleotide repeat length in the HTT gene. Importantly, the CAG repeat undergoes tissue-specific somatic instability, prevalent in brain regions that are disease targets, suggesting a potential role for somatic CAG repeat instability in modifying HD pathogenesis. Thus, understanding underlying mechanisms of somatic CAG repeat instability may lead to discoveries of novel therapeutics for HD. Investigation of the dynamics of the CAG repeat size changes over time may provide insights into the mechanisms underlying CAG repeat instability.
METHODOLOGY/PRINCIPAL FINDINGS: To understand how the HTT CAG repeat length changes over time, we quantified somatic instability of the CAG repeat in Huntington's disease CAG knock-in mice from 2-16 months of age in liver, striatum, spleen and tail. The HTT CAG repeat in spleen and tail was very stable, but that in liver and striatum expanded over time at an average rate of one CAG per month. Interestingly, the patterns of repeat instability were different between liver and striatum. Unstable CAG repeats in liver repeatedly gained similar sizes of additional CAG repeats (approximately two CAGs per month), maintaining a distinct population of unstable repeats. In contrast, unstable CAG repeats in striatum gained additional repeats with different sizes resulting in broadly distributed unstable CAG repeats. Expanded CAG repeats in the liver were highly enriched in polyploid hepatocytes, suggesting that the pattern of liver instability may reflect the restriction of the unstable repeats to a unique cell type.
CONCLUSIONS/SIGNIFICANCE: Our results are consistent with repeat expansion occurring as a consequence of recurrent small repeat insertions that differ in different tissues. Investigation of the specific mechanisms that underlie liver and striatal instability will contribute to our understanding of the relationship between instability and disease and the means to intervene in this process.
亨廷顿病(HD)的发病年龄在很大程度上取决于 HTT 基因中 CAG 三核苷酸重复序列的长度。重要的是,CAG 重复序列经历组织特异性的体细胞不稳定性,在疾病靶标所在的脑区普遍存在,这表明体细胞 CAG 重复序列不稳定性在修饰 HD 发病机制方面可能具有潜在作用。因此,了解体细胞 CAG 重复序列不稳定性的潜在机制可能会发现治疗 HD 的新疗法。研究 CAG 重复序列大小随时间的变化动态可能会深入了解 CAG 重复序列不稳定性的机制。
方法/主要发现:为了了解 HTT CAG 重复序列随时间的变化情况,我们在 2-16 个月大的亨廷顿病 CAG 敲入小鼠的肝脏、纹状体、脾脏和尾巴中定量检测了 CAG 重复序列的体细胞不稳定性。脾脏和尾巴中的 HTT CAG 重复序列非常稳定,但肝脏和纹状体中的 CAG 重复序列随时间推移以每月一个 CAG 的平均速度扩展。有趣的是,肝脏和纹状体中的重复不稳定模式不同。肝脏中不稳定的 CAG 重复序列反复获得相似大小的额外 CAG 重复序列(每月约两个 CAG),维持着独特的不稳定重复序列群体。相比之下,纹状体中不稳定的 CAG 重复序列获得了不同大小的额外重复序列,导致广泛分布的不稳定 CAG 重复序列。肝脏中扩展的 CAG 重复序列高度富集于多倍体肝细胞中,这表明肝脏不稳定的模式可能反映了不稳定重复序列对独特细胞类型的限制。
结论/意义:我们的研究结果与重复序列扩展是作为不同组织中不同的小重复插入的结果一致。研究导致肝脏和纹状体不稳定的具体机制将有助于我们理解不稳定性与疾病之间的关系以及干预这一过程的方法。