Bui Sheryl, Lainé Jeanne, Chevé Maud, Vassilopoulos Stéphane, Lavieu Gregory
Université Paris Cité, NABI, INSERM U1334, UMR 8175/CNRS, 45 Rue Des Saints-Pères, 75006, Paris, France.
Sorbonne Université, INSERM, Institute of Myology, Centre of Research in Myology, UMRS 974, Paris, France.
Sci Rep. 2025 Jun 3;15(1):19454. doi: 10.1038/s41598-025-04576-2.
Extracellular Vesicles (EVs) are natural communication vectors involved in many physiological processes. Significant efforts have aimed to harness EVs for therapeutic delivery, with key challenges being control and enhancement of EV-mediated delivery steps. We and others have developed strategies to improve cargo loading and enhance EV content delivery using viral or non-viral fusogens. However, few targeting solutions have been proposed. Here, we present a versatile system for precise EV targeting to specific cell types, enabling quantitative assessment of targeting efficiency via luminescence and fluorescence. EVs are genetically engineered to express a chimeric adapter protein anchored by a glycosylphosphatidylinositol (GPI) anchor. This protein includes a fluorescent/luminescent domain for detection and a streptavidin domain recruit biotinylated antibodies or ligands specific to cell-surface antigens or receptors. We validated this platform with three different combinations of ligand/target cells, demonstrating up to 40-fold increase in EV uptake. This adaptable system promises to provide a comprehensive solution for targeted therapeutic delivery using EV-based vectors.
细胞外囊泡（EVs）是参与许多生理过程的天然通讯载体。人们付出了巨大努力，旨在利用细胞外囊泡进行治疗性递送，其中关键挑战在于控制和增强细胞外囊泡介导的递送步骤。我们和其他人已经开发出策略，使用病毒或非病毒融合素来改善货物装载并增强细胞外囊泡内容物的递送。然而，很少有靶向解决方案被提出。在这里，我们提出了一个通用系统，用于将细胞外囊泡精确靶向特定细胞类型，能够通过发光和荧光对靶向效率进行定量评估。细胞外囊泡经过基因工程改造，以表达一种由糖基磷脂酰肌醇（GPI）锚定的嵌合衔接蛋白。该蛋白包括一个用于检测的荧光/发光结构域和一个链霉亲和素结构域，可募集针对细胞表面抗原或受体的生物素化抗体或配体。我们用三种不同的配体/靶细胞组合验证了这个平台，证明细胞外囊泡摄取增加了40倍。这个适应性强的系统有望为使用基于细胞外囊泡的载体进行靶向治疗递送提供一个全面的解决方案。
