Dar Ghulam Hassan, Mendes Cláudia C, Kuan Wei-Li, Speciale Alfina A, Conceição Mariana, Görgens André, Uliyakina Inna, Lobo Miguel J, Lim Wooi F, El Andaloussi Samir, Mäger Imre, Roberts Thomas C, Barker Roger A, Goberdhan Deborah C I, Wilson Clive, Wood Matthew J A
Department of Paediatrics, University of Oxford, Oxford, OX1 3QX, UK.
Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, OX1 3QX, UK.
Nat Commun. 2021 Nov 18;12(1):6666. doi: 10.1038/s41467-021-27056-3.
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.
细胞外囊泡（EVs）是在细胞间通讯中起重要作用的生物纳米颗粒，并且具有作为药物递送载体的潜力。在此，我们证明了糖酵解酶甘油醛-3-磷酸脱氢酶（GAPDH）在EV组装和分泌中的作用。我们观察到高水平的GAPDH通过磷脂酰丝氨酸结合基序（G58）与EV的外表面结合，这促进了广泛的EV聚集。在果蝇EV生物发生模型中的进一步研究表明，GAPDH是内体区室中正常生成腔内囊泡所必需的，并促进囊泡聚集。将源自GAPDH的G58肽与双链RNA结合基序融合，能够将小干扰RNA（siRNA）高效加载到EV表面。在亨廷顿舞蹈病小鼠模型中，全身注射后，此类囊泡能有效地将siRNA递送至大脑的多个解剖区域，从而导致大脑不同区域中亨廷顿蛋白基因的沉默。
