Abstract

mRNA-based therapeutics offer significant potential for cancer treatment owing to their ability to induce transient, non-integrating expression of therapeutic proteins. However, effective and tumour-specific delivery remains a major barrier. In this study, we report a cell membrane vesicle (CMV)-based delivery platform for targeted mRNA therapy, employing engineered CMVs enriched with the surface molecule CD6 for the selective delivery of Il24 mRNA to CD166-overexpressing tumour cells. CMVs were derived from NIH-3T3 cells via cytochalasin B induction and genetically modified to express CD6, enabling biomimetic targeting through the CD6–CD166 axis. Il24 mRNA was loaded into CD6-CMVs through a digitonin-assisted permeabilisation strategy, achieving high encapsulation efficiency and sustained intracellular release. In vitro , CD6-CMVs facilitated enhanced uptake in CT26 cells, leading to elevated IL-24 expression, activation of apoptotic and autophagic pathways, and suppression of migration and invasion. In mouse colorectal cancer and squamous carcinoma models, CD6-CMVs/ Il24 mRNA demonstrated effective tumour targeting, increased intratumoral IL-24 translation, and potent antitumor efficacy, with minimal off-target accumulation or systemic toxicity. Notably, this treatment promoted immune cell infiltration and reshaped the tumour microenvironment towards a pro-inflammatory state. This study presents a scalable, low-immunogenicity CMV platform capable of targeted mRNA delivery and cytokine expression, offering a promising strategy for precision immunotherapy for solid tumours.