1. GAPDH controls extracellular vesicle biogenesis and enhances the therapeutic potential of EV mediated siRNA delivery to the brain
- Author
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Roger A. Barker, Clive Wilson, Wooi F. Lim, Alfina A Speciale, Thomas C. Roberts, André Görgens, Samir El Andaloussi, Imre Mäger, Matthew J.A. Wood, Cláudia C. Mendes, Ghulam Hassan Dar, Mariana Conceição, I. Uliyakina, Wei-Li Kuan, Deborah C.I. Goberdhan, Miguel J Lobo, Speciale, Alfina A. [0000-0001-9728-254X], Görgens, André [0000-0001-9198-0857], Lim, Wooi F. [0000-0002-1238-3040], EL Andaloussi, Samir [0000-0003-4468-9113], Mäger, Imre [0000-0003-2242-7227], Roberts, Thomas C. [0000-0002-3313-7631], Goberdhan, Deborah C. I. [0000-0003-0645-6714], Wood, Matthew J. A. [0000-0002-5436-6011], Apollo - University of Cambridge Repository, Speciale, Alfina A [0000-0001-9728-254X], Lim, Wooi F [0000-0002-1238-3040], El Andaloussi, Samir [0000-0003-4468-9113], Roberts, Thomas C [0000-0002-3313-7631], Goberdhan, Deborah CI [0000-0003-0645-6714], Wood, Matthew JA [0000-0002-5436-6011], and Barker, Roger [0000-0001-8843-7730]
- Subjects
Small interfering RNA ,General Physics and Astronomy ,13 ,14 ,59/5 ,59 ,Drug Delivery Systems ,631/61/391/3932 ,42/89 ,RNA, Small Interfering ,13/89 ,Glyceraldehyde 3-phosphate dehydrogenase ,64 ,Huntingtin Protein ,Multidisciplinary ,biology ,Chemistry ,Vesicle ,article ,food and beverages ,Brain ,Glyceraldehyde-3-Phosphate Dehydrogenases ,Extracellular vesicle ,Cell biology ,Huntington Disease ,64/60 ,Protein Binding ,RNAi therapy ,Science ,Phosphatidylserines ,General Biochemistry, Genetics and Molecular Biology ,38 ,Extracellular Vesicles ,stomatognathic system ,Cell Line, Tumor ,Gene silencing ,Animals ,Humans ,Secretion ,Phosphatidylserine binding ,42 ,fungi ,Mesenchymal Stem Cells ,General Chemistry ,Mice, Inbred C57BL ,Disease Models, Animal ,HEK293 Cells ,biology.protein ,Extracellular signalling molecules ,631/80/86/820 ,Biogenesis ,HeLa Cells - Abstract
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., GAPDH is generally considered a housekeeping gene and functions in glycolysis. Here, the authors show that GAPDH has a role in promoting vesicle clustering in endosomes and can load siRNA onto the surface of extracellular vesicles, which can be exploited therapeutically.
- Published
- 2022
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