VPS9D1-AS1 antisense therapy via lipid nanoparticles reprograms cold tumors and enhances immunotherapy in colorectal cancer.

Advanced colorectal cancer (CRC) is characterized by a highly suppressive immune tumor microenvironment (TME) and is insensitive to immunotherapy. The induction of cancer immunogenic cell death (ICD) has been acknowledged as a promising immunotherapeutic strategy. Here, we demonstrate that the long noncoding RNA VPS9D1-AS1 is expressed predominantly in consensus molecular subtype (CMS)-2 CRC, and that its overexpression is associated with immune checkpoint blockade (ICB) sensitivity. A lipid-nanoparticle-(LNP)-based drug delivery system loaded with antisense oligonucleotides (ASOs) was established to target VPS9D1-AS1. This LNP-based drug exhibited high efficiency in inhibiting VPS9D1-AS1 expression in both tumor cells and patient-derived-xenograft (PDX) tumors, significantly suppressing tumor growth and metastasis. Mechanistically, targeting VPS9D1-AS1 activates MLKL-induced ICD, which in turn promotes antigen exposure in tumor cells. Moreover, VPS9D1-AS1 blockade inhibits HLA-G to sensitize CRC cells to immunotherapy. Consequently, LNP-based nano-ASO drugs targeting VPS9D1-AS1 promote TME remodeling by increasing the infiltrations of CD8 T cells and dendritic cells (DCs). In addition, targeting VPS9D1-AS1 harmonizes the crosstalk between tumor cells and DCs via the AXL/GAS6 pathway. In vivo animal studies demonstrated that the combined targeting of PD1 and VPS9D1-AS1 enhances the efficacy of ICB treatment. It is anticipated that nanodrugs targeting VPS9D1-AS1 can be applied in clinical advanced CRC therapy in the near future.