Artificial Transmembrane Oligopeptide‐Integrated Interferon mRNA with Monoclonal Antibody to Improve Specificity and Efficacy of Cancer Immunotherapy

Combination therapy to improve the immunotherapy response rate without causing treatment toxicity remains a significant challenge. Here, an artificial transmembrane oligopeptide, cholesterol-grafted polylysine (CPL), is synthesized to integrate mRNA with a monoclonal antibody into a single system. CPL with a 6% substitution degree is optimized and complexed with IFNβ mRNA to form CmRi nanoparticles. Subsequently, anti-PD-L1 monoclonal antibody (aPD-L1) is modified on the surface of CmRi to produce aCmRi nanoparticles. Notably, CPL facilitates cytosolic delivery via a membrane-interacting mechanism, thereby bypassing lysosomal degradation of mRNA. RNA sequencing analysis revealed that CPL promoted tumor cell apoptosis and upregulated MHC I expression by disrupting lysosomes in tumor cells. Additionally, CPL combined tumor cell-targeting and PD-L1 inhibitory functions of aPD-L1 with the multidimensional immunoadjuvant properties of in situ-expressed IFNβ. This approach overcame the side effects of IFNβ while fully leveraging its therapeutic potential. In a B16-F10 melanoma mouse model, aCmRi demonstrated a superior mRNA transfection rate (26.1%) and achieved the highest tumor suppression rate (89.9%) among all test formulations, without causing significant toxicity. This study presents a simple and versatile strategy to enhance the efficacy, selectivity, and safety of tumor immunotherapy in vivo.