CRISPR/Cas9 Genome Editing Using Lipid Nanoparticles for Hepatocellular Carcinoma Therapy
2Shmunis School of Biomedicine and Cancer Research, George S. Wise Faculty of Life Sciences, Tel Aviv University, Israel
3Department of Materials Sciences and Engineering, Iby and Aladar Fleischman Faculty of Engineering, Tel Aviv University, Israel
4Center for Nanoscience and Nanotechnology, Tel Aviv University, Israel
5Cancer Biology Research Center, Tel Aviv University, Israel
Hepatocellular Carcinoma (HCC) is the 6th most common cancer and one of the most lethal worldwide. According to predictions, HCC mortality will reach 1 million annual deaths by 2030. As of today, HCC remains resistant to most therapies, including new immunotherapy regimens, which commonly achieve only a few months of progression-free survival. Therefore, new therapeutic approaches are necessary, with gene editing being a promising direction. Gene editing is a method in which engineered nucleases are utilized to induce double-strand breaks at specific genomic loci to introduce genomic modifications. One of the most common nucleases currently used in gene editing is the CRISPR/Cas9 system. In the case of cancer, CRISPR/Cas9 can be used to knock out a critical gene and lead to cancerous cells` death. In this work, the targeted gene is NCAPG which is a part of the chromosome condensation process and has been found important in various cancers including HCC. For this purpose, several sgRNAs were screened to achieve the highest rate of NCAPG editing with the best achieving ~85% editing. Further testing showed that NCAPG editing leads to ~60% mortality in HCC cells. We are currently screening various lipid nanoparticle formulations to co-deliver Cas9 mRNA alongside the sgRNA. Finally, we are currently in the process of creating a mouse model to test the Cas9 efficacy in-vivo. It is our belief that this CRISPR/Cas9-based approach may open a new avenue for treating HCC.