Picosecond Pulsed Electric Field-Induced Disaggregation of Polyglutamine Aggregates in Huntington’s Disease Neural Stem Cells

Huntington’s disease (HD) is a neurodegenerative disorder marked by polyglutamine (PolyQ) aggregation and mitochondrial dysfunction, yet non-invasive methods to modulate these intracellular stressors remain limited. Here, we applied picosecond pulsed electric fields (psPEF)—an ultrashort bioelectronic modality—to patient-derived HD neural stem cells (NSCs) to assess changes in pathogenic protein aggregation, gene expression, and mitochondrial membrane potential. Using a custom 3D bioprinter-based stimulation platform, cells were exposed to non-contact electric fields of 20 or 40 kV/cm with subnanosecond pulse width (660 ps). Quantitative imaging and automated analysis revealed a significant reduction in aggregate size and aggresome burden within 30 minutes post-treatment, effects that persisted at 24 hours without compromising viability. HTT mRNA levels remained unchanged, supporting a post-translational mechanism of aggregate modulation. We also observed a transient redistribution of aggregates into the nuclear compartment and a field-dependent trend toward increased mitochondrial polarization, suggestive of broader proteostatic or bioenergetic effects. Transcript analysis revealed downregulation of PAX6 and CACNA1C, further implicating psPEF in modulating intracellular stress pathways. These findings represent the first evidence that ultrashort electric fields can reduce mutant HTT aggregation in a human HD model without genetic manipulation or membrane poration. Our results establish HD-NSCs as a scalable, disease-relevant platform for evaluating psPEF in neurodegenerative disease and support further exploration of dielectric mechanisms for intracellular remodeling. Collectively, this work introduces a contactless, non-invasive strategy for modulating protein aggregation and mitochondrial stress in human neural cells, offering a new direction for therapeutic development in proteopathy-driven conditions.