Enhancing CRISPR/Cas gene editing through modulating cellular mechanical properties for cancer therapy

Genome editing holds great potential for cancer treatment due to the ability to precisely inactivate or repair cancer-related genes. However, delivery of CRISPR/Cas to solid tumors for efficient cancer therapy remains challenging. Here, we targeted tumor tissue mechanics via a multiplexed dendrimer lipid nanoparticle (LNP) approach involving co-delivery of focal adhesion kinase (FAK) siRNA, Cas9 mRNA, and sgRNA (siFAK+CRISPR-LNPs) to enable tumor delivery and enhance gene editing efficacy. We show that gene editing was enhanced >10-fold in tumor spheroids due to increased cellular uptake and tumor penetration of nanoparticles mediated by FAK-knockdown. siFAK+CRISPR-PD-L1-LNPs reduced extracellular matrix stiffness and efficiently disrupted PD-L1 expression by CRISPR/Cas gene editing, which significantly inhibited tumor growth and metastasis in four mouse models of cancer. Overall, we provide evidence that modulating the stiffness of tumor tissue can enhance gene editing in tumors, which offers a new strategy for synergistic LNPs and other nanoparticle systems to treat cancer using gene editing.