Portal de Periódicos da CAPES

Huntingtin CAG-expansion mutation results in a dominant negative effect

2023; Frontiers Media; Volume: 11; Linguagem: Inglês

10.3389/fcell.2023.1252521

2296-634X

Tiago L. Laundos, Shu Li, Eric Cheang, Riccardo De Santis, Francesco M. Piccolo, Ali H. Brivanlou,

RNA Research and Splicing

Huntington's disease (HD) remains an incurable and fatal neurodegenerative disease long after CAGexpansion mutation in the huntingtin gene (HTT) was identified as the cause. The underlying pathological mechanism, whether HTT loss of function or gain of toxicity results from mutation, remains a matter of debate. In this study, we genetically modulated wild-type or mutant HTT expression levels in isogenic human embryonic stem cells to systematically investigate their contribution to HDspecific phenotypes. Using highly reproducible and quantifiable in vitro micropattern-based assays, we observed comparable phenotypes with HD mutation and HTT depletion. However, halving endogenous wild-type HTT levels did not strongly recapitulate the HD phenotypes, arguing against a classical loss of function mechanism. Remarkably, expression of CAG-expanded HTT in non-HD cells induced HDlike phenotypes akin to HTT depletion. By corollary, these results indicate a dominant negative effect of mutated HTT on its wild-type counterpart. Complementation with additional copies of wild-type 2 This is a provisional file, not the final typeset article HTT ameliorated the HD-associated phenotypes, strongly supporting a classical dominant negative mechanism. Understanding the molecular basis of this dominant negative effect will guide the development of efficient clinical strategies to counteract the deleterious impact of mutant HTT on the wild-type proteinHTT function.