Department of Biology, Boston University, Boston, MA 02215, USA.
Department of Bioengineering, College of Engineering, Hanyang University, Seoul 04763, Republic of Korea.
J Control Release. 2021 Feb 10;330:684-695. doi: 10.1016/j.jconrel.2020.12.053. Epub 2021 Jan 1.
Exosomes have been investigated as delivery vesicles for various drugs. However, exosome-mediated peptide delivery into the lungs has not been studied. In this study, exosomes were engineered for the pulmonary delivery of RAGE-binding peptide (RBP), an anti-inflammatory peptide. To load the peptide into exosomes, RBP was linked to an exosome membrane integral protein, Lamp2b, to produce RBP-linked exosomes (RBP-exo). The anti-inflammatory effects of RBP-exo were confirmed by cytokine assays in lipopolysaccharide (LPS)-activated macrophage cells. To increase anti-inflammatory effects, curcumin was loaded into RBP-exo. Curcumin loaded RBP-exo (RBP-exo/Cur) had higher intracellular curcumin delivery efficiency than curcumin alone or curcumin loaded into unmodified exosomes (unmod-exo/Cur). This suggests that RBP on the surface of RBP-exo may interact with RAGE and increase the intracellular delivery efficiency of curcumin. In addition, RBP-exo/Cur had higher anti-inflammatory effects than curcumin alone, a mixture of RBP and curcumin, and unmod-exo/Cur in vitro. For in vivo evaluation, RBP-exo/Cur was administrated by intratracheal instillation into the lungs of an acute lung injury (ALI) model. The results showed that RBP-exo/Cur reduced pro-inflammatory cytokines more efficiently than curcumin alone, RBP-exo, and unmod-exo/Cur. Hematoxylin and eosin staining confirmed that the inflammation reaction was inhibited in the RBP-exo and RBP-exo/Cur groups. Immunostaining assays showed that RBP-exo was co-localized mostly with type I epithelial cells. In conclusion, RBP was successfully delivered with exosomes into the lungs by inhalation. A combination of RBP and curcumin using exosomes as carriers may be useful as ALI therapy.
外泌体已被研究作为各种药物的递药载体。然而，外泌体介导的肽递送至肺部尚未得到研究。在这项研究中，工程化的外泌体用于肺部递送达肽（RBP），一种抗炎肽。为了将肽装载到外泌体中，将 RBP 连接到外泌体膜整合蛋白 Lamp2b 上，以产生 RBP 连接的外泌体（RBP-exo）。通过脂多糖（LPS）激活的巨噬细胞细胞中的细胞因子测定来证实 RBP-exo 的抗炎作用。为了增加抗炎作用，将姜黄素载入 RBP-exo。载有姜黄素的 RBP-exo（RBP-exo/Cur）比单独的姜黄素或载入未修饰的外泌体（unmod-exo/Cur）中的姜黄素具有更高的细胞内姜黄素递送效率。这表明 RBP-exo 表面的 RBP 可能与 RAGE 相互作用，从而提高姜黄素的细胞内递送效率。此外，RBP-exo/Cur 在体外具有比单独的姜黄素、RBP 和姜黄素的混合物以及 unmod-exo/Cur 更高的抗炎作用。为了进行体内评价，通过气管内滴注将 RBP-exo/Cur 递送至急性肺损伤（ALI）模型的肺部。结果表明，与单独的姜黄素、RBP-exo 和 unmod-exo/Cur 相比，RBP-exo/Cur 更有效地减少促炎细胞因子。苏木精和伊红染色证实 RBP-exo 和 RBP-exo/Cur 组抑制了炎症反应。免疫染色测定表明 RBP-exo 主要与 I 型上皮细胞共定位。总之，通过吸入成功地将 RBP 与外泌体一起递送至肺部。使用外泌体作为载体的 RBP 和姜黄素的组合可能对 ALI 治疗有用。
