Nanoparticle Orientation to Control RNA Loading and Ligand Display on Extracellular Vesicles for Cancer Regression

Nanotechnology holds many advantages. Here we report another advantage of applying RNA nanotechnology for directional control. The orientation of arrow-shaped RNA was altered to control ligand-display on extracellular vesicle (EV) membranes for specific cell targeting, or to regulate intracellular trafficking of siRNA/miRNA. Placing membrane-anchoring cholesterol at the arrow-tail results in display of RNA aptamer or folate on EV outer surface. In contrast, placing the cholesterol at the arrow-head results in partial loading of RNA nanoparticles into the EVs. Taking advantage of the RNA ligand for specific targeting and EVs for efficient membrane fusion, the resulting ligand-displaying EVs were competent for specific delivery of siRNA to cells, and efficiently block tumor growth in three cancer models. PSMA aptamer-displaying EVs loaded with survivin siRNA inhibited prostate cancer xenograft. The same EV but displaying EGFR aptamer inhibited orthotopic breast cancer models. Likewise, survivin-loaded and folate-displaying EVs inhibited patient derived colorectal cancer xenograft.