An erythrocyte membrane-fused plant-derived nanoparticles as a gene therapy vehicle for the treatment of CI/R injury

Ischemic stroke is currently the second leading cause of death worldwide, and insufficient endogenous neurogenesis is the greatest cause of post-stroke disability. MicroRNAs have been proven to hold therapeutic potential, unfortunately, they have a low stability that hinders their clinical usage. Our earlier work revealed that Panax notoginseng derived exosome like nanoparticles, namely PDNs have potential to bypass BBB and reduce the cerebral ischemia/reperfusion (CI/R) damage. In this study, we employed microRNA-124 as a model therapeutic gene, utilizing its engineered variant Agomir-124 (Ago124) to optimize loading efficiency. The therapeutic effects of Ago124@R-PDN were further assessed in several sets of experiments. Pharmacokinetic study showed that erythrocyte membrane extended the half-life of PDNs from 7 min to 11.3 h, and the loading efficiency of Ago124 reached 40 %. In an <i>in vitro</i> oxygen-glucose deprivation/reperfusion (OGD/R) model, Ago124@R-PDN enhanced IL-10 production in microglia by 67 % (<i>vs</i> 11.7 % with free Ago124), and promoted Tuj1⁺ neuronal differentiation by 2.23-fold compared with vehicle. Also, Ago124@R-PDN brought gene cargo into the brain, alleviated infarct volume, and improved functional behaviors in model mice. At last, we demonstrated that surface glycosyl of PDN facilitated its brain-entering ability by being recognized by sodium-glucose linked transporter-1 protein. In conclusion, our erythrocyte fused PDNs offer a promising strategy for delivering biomacromolecule to treat brain diseases.