Enhanced Small Extracellular Vesicle Uptake By Activated Interneurons Improves Stroke Recovery In Mice
Recommended Citation
Wang M, Liu Z, Chopp M, Millman M, Li Y, Cepparulo P, Li C, Kemper A, Zhang L, Zhang Z, Zhang Y. Enhanced Small Extracellular Vesicle Uptake By Activated Interneurons Improves Stroke Recovery In Mice. Stroke 2025; 56.
Document Type
Conference Proceeding
Publication Date
1-30-2025
Publication Title
Stroke
Keywords
adenosine triphosphate, amine, clozapine n oxide, dextran, transcription factor, Adeno associated virus, adult, animal cell, animal experiment, animal model, animal tissue, axon, brain cell, cerebrovascular accident, chemogenetics, conference abstract, controlled study, endothelium cell, exosome, female, Golgi stain, infarct volume, interneuron, male, middle cerebral artery occlusion, mouse, nerve cell network, nonhuman, oxidative phosphorylation, pharmacology, pyramidal tract, tomato, transmission electron microscope, Western blotting
Abstract
Background: Neuronal circuitry remodeling which compromises excitatory and inhibitory neurons is critical to improve neurological outcome after stroke. Cerebral endothelial cell (CEC) generated small extracellular vesicles (CEC-sEVs) have a therapeutic effect on stroke recovery. However, it remains challenge to use sEVs for specifically targeting individual neurons for enhancement of the circuitry after stroke. Using a chemogenetic approach, we tested the hypothesis that Designer Receptors Exclusively Activated by Designer Drugs (DREADD) activated peri-infarct cortical interneurons preferentially take up exogenous CEC-sEVs to enhance neuronal remodeling and functional recovery. Methods: Adeno-associated viruses (AAVs) carrying Gq-DREADD and Tomato under the control of the transcription factor distal-less homeobox (Dlx) enhancer element (AAV-hDlx-Gq, 500 nL) were injected into the cortex. The mice were then subjected to middle cerebral artery occlusion (MCAO). CEC-sEVs (1x10 particles, i.v.) were administered following interneuron activation by Clozapine-N-oxide (CNO, 5mg/kg, i.p). The circuitry was assayed by Golgi-Cox staining and biotinylated dextran amine labeled axons in the corticospinal tract (CST). Transmission electron microscope (TEM) was used to measure CEC-sEVs in neurons and neuronal mitochondrial (Mito) integrity. Results: The adjunct therapy of CEC-sEVs and CNO increased CEC-sEV uptake by peri-infarct cortical neurons and improved somatosensory functional outcome by ~70% vs CEC-sEVs or CNO alone (p<0.05, n=10/group). The adjunct therapy (p<0.05) augmented peri-infarct cortical axonal/dendritic outgrowth and CST axonal remodeling but did not reduce infarct volume. TEM analysis revealed that the neurons with uptake of CEC-sEVs (p<0.05) increased their Mito with intact cristae. Western blot showed that adjunct CEC-sEVs with CNO (p<0.05) decreased ischemia-increased Drp1 and Mff, and elevated ischemia-reduced Mito oxidative phosphorylation (OXPHOS) protein, COX5A, vs CEC-sEVs or CNO alone. Increases of Mito Drp1 and Mff impair Mito crista integrity, which reduces OXPHOS-induced ATP production. Conclusions: Our data demonstrate that activated cortical interneurons preferentially increased the uptake of CEC-sEVs, leading to the improvement of functional outcome after stroke. The increased CEC-sEV uptake by activated interneurons reduced ischemic neuronal Mito damage, which may contribute to enhanced neuronal circuitry remodeling.
Volume
56
