Fig. 1: Schematic design of E. coli protoplast-derived nanovesicles (sgPik3cg-DHP/DGA-NVs) for TAM-selective genome editing to enhance anti-tumor efficacy. | Nature Communications

Fig. 1: Schematic design of E. coli protoplast-derived nanovesicles (sgPik3cg-DHP/DGA-NVs) for TAM-selective genome editing to enhance anti-tumor efficacy.

From: Bacterial protoplast-derived nanovesicles carrying CRISPR-Cas9 tools re-educate tumor-associated macrophages for enhanced cancer immunotherapy

Fig. 1

In this illustration, we outline the creation of sgPik3cg-DHP/DGA-NVs for targeted genome editing in tumor-associated macrophages (TAMs) to enhance anti-tumor effects. The process begins with the construction of E. coli expressing the Cas9-sgPik3cg complex. Subsequently, the bacterial outer membrane, which possesses high endotoxicity, is removed. This results in the formation of sgPik3cg-DHP/DGA-NVs, which encapsulate a substantial amount of Cas9-sgPik3cg ribonucleoproteins (RNPs) and CpG-rich genomic DNA. These nanovesicles (NVs) are produced through a series of extrusion steps and are further modified with a pH-responsive phospholipid derivative (DHP) and a phospholipid derivative targeted specifically to TAM (DGA). Upon intravenous injection, sgPik3cg-DHP/DGA-NVs accumulate in tumor tissues due to their prolonged circulation capability and the enhanced permeability and retention (EPR) effect. Within the acidic microenvironment of the tumor, PEG2000 separates from DHP, triggering the recognition and internalization of DGA-functionalized NVs by TAMs via macrophage galactose-type lectin (MGL) receptor-mediated endocytosis. This process enables TAM-specific genome editing of Pik3cg and activation of toll-like receptor 9 (TLR9) in vivo, resulting in the reprogramming of M2-like TAMs into an anti-tumor M1-like phenotype and facilitating tumor immunotherapy.

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