Therapeutic Genome editing in Cancer via Targeted Lipid Nanoparticles entrapping RNAs

Harnessing editing technologies for cancer therapeutics has been hampered by low editing efficiency in tumors, lack of selective delivery to tumors and potential toxicity of the existing delivery systems. In my presentation I will describe a safe and efficient lipid-nanoparticle (LNP) for the delivery of Cas9 mRNA and sgRNAs that utilize novel amino-ionizable lipids with cell specific targeting strategy.
I will detailed several examples including: a single intracerebral injection of CRISPR-LNPs against PLK1 (sgPLK1-cLNPs) into aggressive orthotopic glioblastoma, enabled up to ~ 70% gene editing in-vivo, which caused tumor-cells apoptosis, inhibited tumor-growth by 50%, and improved survival in mice by 30%.
To reach disseminated tumors, cLNPs were also engineered for antibody-targeted delivery. Intraperitoneal injections of EGFR-targeted sgPLK1-cLNPs caused their selective uptake into disseminated ovarian tumors, enabeled up to ~80% gene-editing in-vivo, inhibited tumor-growth, and increased survival in tumor-bearing mice by 80%.
I will also introduce a safe and effective intratumoral EGFR-targeted CRISPR-LNP delivery strategy for knocking out SOX2, which is a cancer-specific gene. To assess their therapeutic potential, we showed that LNPs made from ionizable lipids with helper lipids co-encapsulating Cas9 mRNA and sgRNA targeting SOX2 (sgSOX2), lead to a ~60% reduction in HNSCC cell viability in vitro. Next, using a xenograft of Squamous cell carcinomas of the head and neck (HNSCC) mouse model, targeted delivery of 𝜶EGFR- CRISPR-sgSOX2-LNPs to HNSCC cells resulted in a 90% inhibition of tumor growth and a 90% increase in survival for > 84 days, with tumor disappearance observed in 50% of the mice. The ability to disrupt gene expression in-vivo in extrahepatic tumors and in a cell specific manner opens new avenues for cancer therapeutics.