Extracellular vesicles (EVs) are biological nanoparticles with important roles in intercellular communication, and potential as drug delivery vehicles. Here we demonstrate a role for the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in EV assembly and secretion. We observe high levels of GAPDH binding to the outer surface of EVs via a phosphatidylserine binding motif (G58), which promotes extensive EV clustering. Further studies in a Drosophila EV biogenesis model reveal that GAPDH is required for the normal generation of intraluminal vesicles in endosomal compartments, and promotes vesicle clustering. Fusion of the GAPDH-derived G58 peptide to dsRNA-binding motifs enables highly efficient loading of small interfering RNA (siRNA) onto the EV surface. Such vesicles efficiently deliver siRNA to multiple anatomical regions of the brain in a Huntington's disease mouse model after systemic injection, resulting in silencing of the huntingtin gene in different regions of the brain.
Animals, Brain, Cell Line, Tumor, Disease Models, Animal, Drug Delivery Systems, Extracellular Vesicles, Glyceraldehyde-3-Phosphate Dehydrogenases, HEK293 Cells, HeLa Cells, Humans, Huntingtin Protein, Huntington Disease, Mesenchymal Stem Cells, Mice, Inbred C57BL, Phosphatidylserines, Protein Binding, RNA, Small Interfering