A compact GAD67 promoter enables inhibitory neuron-targeted AAV gene therapy for seizure suppression

Abstract

Epilepsy arises from disruption of excitation-inhibition (E/I) balance, typically due to excessive excitatory activity. Despite available therapies, a substantial proportion of patients remain treatment resistant. Enhancing inhibitory neuron activity via gene therapy can restore E/I balance and may therefore provide a therapeutic strategy for treatment-resistant epilepsy. Here, we developed a compact 410-bp glutamic acid decarboxylase 67 promoter (cmGAD67) that enables strong, selective transgene expression in inhibitory neurons while preserving adeno-associated virus (AAV) packaging capacity. Systemic delivery of AAV vectors carrying cmGAD67 preferentially targeted parvalbumin interneurons and enabled efficient circuit modulation. To evaluate therapeutic potential, we expressed glutamic acid decarboxylase 65 (GAD65) under the control of cmGAD67 (AAV-GAD65). AAV-GAD65 suppressed abnormal delta oscillations, reduced seizure-like activity, normalized anxiety-like behavior, and improved survival in seizure models. Biochemical analyses confirmed increased GABA levels in the cortex and hippocampus, linking functional improvements to enhanced inhibitory neurotransmitter synthesis. Together, these findings establish the cmGAD67 promoter as a versatile platform for inhibitory neuron-targeted AAV gene delivery and identify AAV-GAD65 as a promising strategy for seizure control and disorders associated with E/I imbalance.

Abstract of "A compact GAD67 promoter enables inhibitory neuron-targeted AAV gene therapy for seizure suppression" by Fukai Y, Konno A, Hosoi N et al., https://doi.org/10.1016/j.ymthe.2026.06.007, used under its Creative Commons Attribution license (CC-BY); markup removed, otherwise unmodified.