Targeted Delivery of CRISPR/Cas9‐Mediated Cancer Gene Therapy via Liposome‐Templated Hydrogel Nanoparticles

TLDR

CRISPR/Cas9 is a highly efficient gene‑editing platform for cancer therapy, yet safe and specific delivery to tumors remains a critical barrier. The study reports a novel liposome‑templated hydrogel nanoparticle (LHNP) core‑shell design engineered for efficient co‑delivery of Cas9 protein and guide RNA. LHNPs consist of a liposome core encapsulating Cas9 and nucleic acids within a hydrogel shell, enabling controlled release and tumor targeting. In vitro and in vivo experiments show that LHNPs deliver CRISPR/Cas9 more efficiently than Lipofectamine 2000, enable targeted inhibition of genes such as PLK1 in brain and other tumors, and significantly suppress tumor growth and extend survival in mouse models, indicating their potential as a versatile cancer gene‑therapy platform.

Abstract

Due to its simplicity, versatility, and high efficiency, the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 technology has emerged as one of the most promising approaches for treatment of a variety of genetic diseases, including human cancers. However, further translation of CRISPR/Cas9 for cancer gene therapy requires development of safe approaches for efficient, highly specific delivery of both Cas9 and single guide RNA to tumors. Here, novel core-shell nanostructure, liposome-templated hydrogel nanoparticles (LHNPs) that are optimized for efficient codelivery of Cas9 protein and nucleic acids is reported. It is demonstrated that, when coupled with the minicircle DNA technology, LHNPs deliver CRISPR/Cas9 with efficiency greater than commercial agent Lipofectamine 2000 in cell culture and can be engineered for targeted inhibition of genes in tumors, including tumors the brain. When CRISPR/Cas9 targeting a model therapeutic gene, polo-like kinase 1 (PLK1), is delivered, LHNPs effectively inhibit tumor growth and improve tumor-bearing mouse survival. The results suggest LHNPs as versatile CRISPR/Cas9-delivery tool that can be adapted for experimentally studying the biology of cancer as well as for clinically translating cancer gene therapy.