Optimization of trans-Splicing for Huntington's Disease RNA Therapy

Frontiers in Neuroscience

Virginia B. Mattis¹  Colton M. Tom¹  Christian L. Lorson²  Hansjörg Rindt² 

[1] Cedars-Sinai Medical Center, Board of Governors Regenerative Medicine Institute, Los Angeles, CA, United States;Department of Veterinary Pathobiology, Bond Life Sciences Center, University of Missouri, Columbia, MO, United States;

关键词: Huntington's disease;    HD;    Huntingtin;    HTT;    trans-splicing;    therapy;   

DOI  :  10.3389/fnins.2017.00544

来源: DOAJ

【 摘 要 】

Huntington's disease (HD) is a devastating neurodegenerative disorder caused by a polyglutamine (polyQ) expansion in exon 1 of the Huntingtin (HTT) gene. We have previously demonstrated that spliceosome-mediated trans-splicing is a viable molecular strategy to specifically reduce and repair mutant HTT (mtHTT). Here, the targeted tethering efficacy of the pre-mRNA trans-splicing modules (PTM) in HTT was optimized. Various PTMs that targeted the 3′ end of HTT intron 1 or the intron 1 branch point were shown trans-splice into an HTT mini-gene, as well as the endogenous HTT pre-mRNA. PTMs that specifically target the endogenous intron 1 branch point increased the trans-splicing efficacy from 1–5 to 10–15%. Furthermore, lentiviral expression of PTMs in a human HD patient iPSC-derived neural culture significantly reversed two previously established polyQ-length dependent phenotypes. These results suggest that pre-mRNA repair of mtHTT could hold therapeutic benefit and it demonstrates an alternative platform to correct the mRNA product produced by the mtHTT allele in the context of HD.

【 授权许可】

Unknown