Autonomic lysosomal escape via sialic acid modification enhances mRNA lipid nanoparticles to eradicate tumors and build humoral immune memory.

Lysosomes present a major barrier to efficient mRNA delivery. Existing strategies primarily depend on lysosomal disruption, which is inefficient and carries a risk of cytolysis. We propose an Autonomic Lysosomal Escape (ALE) strategy, in which sialic acid (SA) modification enables over 90 % of LNPs to successfully escape from lysosomes by inducing cells to spontaneously reduce lysosome generation. The SA modification enhances the transfection efficiency of LNPs administered via intravenous injection, intramuscular injection, and inhalation, demonstrating the broad applicability. The structure of cleavable PEG-lipids was optimized using a newly developed method, termed Systematic Evaluation of LNPs' Efficiency by Cumulative Tests (SELECT). The results showed that polyethylene glycol 2000-cholesterol hemisuccinate (Ps) is the optimal candidate for co-modification with SA. The resulting LNPs co-modified with SA and Ps (SAPs@LNPs) completely eradicated TC-1 tumors and induced humoral immune memory. Combining SA-modified doxorubicin liposomes (DOX-SL) further accelerates tumor elimination, while licensed PEGylated liposomal doxorubicin (Caelyx) impairs the efficacy of mRNA vaccines. This difference stems from DOX-SL's selective depletion of tumor-associated immune cells (TAICs) and the nonspecific cytotoxicity of Caelyx. These findings suggest that combining Caelyx with mRNA vaccines should be approached with caution. Our study also highlights the key roles of humoral immune memory and natural killer cell-driven antibody-dependent cellular cytotoxicity (ADCC) in tumor eradication, and incorporating them into the cancer immune cycle further refines this theory.