Structural insight into transmissive mutant huntingtin species by correlative light and electron microscopy and cryo-electron tomography

Aggregates of mutant huntingtin (mHTT) containing an expanded polyglutamine (polyQ) tract are hallmarks of Huntington's Disease (HD). Studies have shown that mHTT can spread between cells, leading to the propagation of misfolded protein pathology. However, the structure of transmissive mHTT species, and the molecular mechanisms underlying their transmission remain unknown. Using correlative light and electron microscopy (CLEM) and cryo-electron tomography (cryo-ET), we identified two types of aggregation-prone granules in conditioned medium from PC12 cells expressing a mHTT N-terminal fragment: densities enclosed by extracellular vesicles (EVs), and uncoated, amorphous meshworks of heterogeneous oligomers that co-localize with clusters of EVs. In vitro assays confirmed that liposomes induce condensation of polyQ oligomers into higher-order assemblies, resembling the uncoated meshworks observed in PC12 conditioned medium. Our findings provide novel insights into formation and architecture of transmissive mHTT proteins, and highlight the potential role of EVs as both carriers and modulators of transmissive mHTT proteins. •Mutant huntingtin spreads between cells leading to misfolded protein propagation.•Transmissive mutant huntingtin exists in both vesicle-enclosed and freestanding forms.•Extracellular vesicles serve as carriers and structural modulators of transmissive mutant huntingtin.•Cutting-edge bioimaging of polymorphic transmissive structures in cellular environment.