Hierarchical ZIF-8@Au cluster Nanocarriers for In Situ CAR-M programming and Immunotherapy in Prostate Cancer

Here, we have successfully constructed a novel nano-delivery system based on the Zeolitic Imidazolate Framework-8 (ZIF-8), which revealed high CAR gene encapsulation efficiency, reduced non-specific hepatic accumulation, targeted delivery of tumor-associated macrophages (TAM) and efficient intracellular gene transfection efficiency enable the in-situ construction of CAR-M cells. Meanwhile, the co-delivery of the IFN-γ and CAR genes not only maintains the specific killing and phagocytic activity of CAR-M against tumor cells but also activates the adaptive immunity, inducing excellent anti-tumor efficacy in a prostate cancer mouse model. In summary, this strategy provides a novel approach for the systematic in vivo editing of CAR-M. The obtained bone marrow-derived macrophages were transfected with PCI plasmid and PC plasmid via our nanocarriers, respectively, and then incubated with the cell incubator at 37 ° C for 48h and then collect these cells. There were two parallel experimental groups in each group.The RNA was extracted by Trizol method, and the library was constructed using Fast Rna-Seq Lib Prep Kit V2 (Cat.No.RK20306). Transcriptome related data of these macrophages were generated by deep sequencing, in triplicate, using Illumina NovaSeq X Plus. RNA samples were subjected to rigorous quality control through an Agilent 2100 bioanalyzer. PCI plasmid structure include PSMA-CAR-IFN-γ, while PCI plasmid structure just include PSMA-CAR. PSMA is the abbreviation of prostate cancer-specific antigen, CAR is the abbreviation of chimeric antigen receptor, IFN-γ is the abbreviation of interferon-γ.