Matuszek Zaneta, Arbab Mandana, Kesavan Maheswaran, Hsu Alvin, Roy Jennie C L, Zhao Jing, Yu Tian, Weisburd Ben, Newby Gregory A, Doherty Neil J, Wu Muzhou, Shibata Shota, Cristian Ana, Tao Y Allen, Fearnley Liam G, Bahlo Melanie, Rehm Heidi L, Xie Jun, Gao Guangping, Mouro Pinto Ricardo, Liu David R
Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of Harvard and MIT, Cambridge, MA, USA.
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.
Nat Genet. 2025 May 26. doi: 10.1038/s41588-025-02172-8.
Trinucleotide repeat (TNR) diseases are neurological disorders caused by expanded genomic TNRs that become unstable in a length-dependent manner. The CAG•CTG sequence is found in approximately one-third of pathogenic TNR loci, including the HTT gene that causes Huntington's disease. Friedreich's ataxia, the most prevalent hereditary ataxia, results from GAA repeat expansion at the FXN gene. Here we used cytosine and adenine base editing to reduce the repetitiveness of TNRs in patient cells and in mice. Base editors introduced G•C>A•T and A•T>G•C interruptions at CAG and GAA repeats, mimicking stable, nonpathogenic alleles that naturally occur in people. AAV9 delivery of optimized base editors in Htt.Q111 Huntington's disease and YG8s Friedreich's ataxia mice resulted in efficient editing in transduced tissues, and significantly reduced repeat expansion in the central nervous system. These findings demonstrate that introducing interruptions in pathogenic TNRs can mitigate a key neurological feature of TNR diseases in vivo.
三核苷酸重复(TNR)疾病是由基因组TNR扩增引起的神经疾病,这些TNR会以长度依赖的方式变得不稳定。约三分之一的致病性TNR位点中存在CAG•CTG序列,包括导致亨廷顿舞蹈病的HTT基因。最常见的遗传性共济失调——弗里德赖希共济失调,是由FXN基因处的GAA重复扩增导致的。在此,我们利用胞嘧啶和腺嘌呤碱基编辑来降低患者细胞和小鼠中TNR的重复次数。碱基编辑器在CAG和GAA重复序列处引入G•C>A•T和A•T>G•C干扰,模拟人类中自然存在的稳定、无致病性的等位基因。在Htt.Q111亨廷顿舞蹈病小鼠和YG8s弗里德赖希共济失调小鼠中通过AAV9递送优化的碱基编辑器,在转导组织中实现了高效编辑,并显著减少了中枢神经系统中的重复扩增。这些发现表明,在致病性TNR中引入干扰可在体内减轻TNR疾病的关键神经特征。