Monitoring huntingtin aggregation in Huntington's disease: methods, challenges and future opportunities

Huntington's disease is characterized by the progressive misfolding and aggregation of mutant huntingtin, a dynamic process generating a heterogeneous spectrum of soluble oligomers, fibrillar assemblies and inclusion bodies. Increasing evidence suggests that distinct aggregation intermediates differ in their toxicity and biological function, making accurate detection of these species essential for understanding disease mechanisms and for developing effective therapeutic interventions.

In this review, we evaluate methodologies currently used to study mutant huntingtin aggregation, including biochemical and imaging approaches. The literature search for peer-reviewed articles up to 2026 was conducted using Pubmed and Google Scholar, with a preference for more recent papers. Based on this search, we discuss which stages of the aggregation pathway are captured and their strengths and limitations. We highlight that methodology strongly shapes observations, as each approach preferentially detects specific aggregate species, necessitating complementary strategies.

We propose future opportunities to develop minimally perturbative screening platforms by combining advances in intrabody engineering with high-content imaging and single-cell analytical approaches. Such integrated strategies could enable comprehensive detection of the full spectrum of mHTT aggregation while simultaneously linking distinct aggregation states to multi-omic signatures in physiologically relevant cellular models.