Activation of RIG‐I/MDA5 Signaling and Inhibition of CD47‐SIRPα Checkpoint with a Dual siRNA‐Assembled Nanoadjuvant for Robust Cancer Immunotherapy

Abstract Antigen‐presenting cells (APCs) play a crucial role in the anti‐tumor immunity as they are responsible for capturing, processing, and presenting tumor antigens to T cells. However, their activation is often limited by the absence of adjuvants and the suppressive effects of immune checkpoints, such as CD47‐SIRPα. Herein, we present a nanoadjuvant that is self‐assembled from long RNA building blocks generated through rolling circle transcription (RCT) reaction and further modified with cationic liposomes. Owing to the high load of densely packed RNA, this nanoadjuvant could robustly activate RIG‐I/MDA5 signaling in APCs, leading to the maturation of dendritic cells (DCs) and the polarization of tumor‐associated macrophages (TAMs) toward an anti‐tumor M1‐like phenotype. In addition, with a well‐designed template, the generated long RNA from RCT reaction includes two kinds of siRNA targeting both CD47 in tumor cells and SIRPα in APCs. This dual gene silencing results in efficient inhibition of the CD47‐SIRPα checkpoint. Collectively, the robust activation of RIG‐I/MDA5 signaling and efficient inhibition of CD47‐SIRPα checkpoint enhance the phagocytic activity of APCs, which in turn promotes the cross‐priming of effector T cells and the activation of anti‐tumor immune responses. This study therefore provides a simple and robust RNA nanoadjuvant for cancer immunotherapy.