Charge-reversal nanocomolexes-based CRISPR/Cas9 delivery system for loss-of-function oncogene editing in...

Area of Science:

• Biomedical Engineering
• Nanotechnology
• Oncology

Background:

• Hepatocellular carcinoma (HCC) presents significant global health challenges due to high mortality and treatment resistance.
• Current chemotherapy and targeted therapies for HCC face limitations, necessitating novel treatment strategies.

Purpose of the Study:

• To develop and evaluate a novel charge-reversal nanoparticle system (Hep@PGEA) for delivering CRISPR-Cas9 gene editing components to treat HCC.
• To assess the in vitro and in vivo anti-tumor efficacy of the Hep@PGEA/pCas9 system against HCC.

Main Methods:

• Fabrication of a responsive charge-reversal vehicle (Hep@PGEA) with a heparin core and EA-modified PGEA shell.
• Encapsulation of pCas9 plasmid and sgRNA targeting survivin within the Hep@PGEA vehicle.
• In vitro evaluation of anti-tumor activity, including apoptosis induction and inhibition of proliferation, migration, and invasion.
• In vivo studies using an orthotopic HCC mouse model to assess liver accumulation and therapeutic effects, including combination therapy with sorafenib.

Main Results:

• The Hep@PGEA/pCas9 system demonstrated potent anti-tumor efficiency in vitro by inducing apoptosis and suppressing HCC cell growth and metastasis.
• In vivo administration resulted in significant accumulation within the liver and notable anti-tumor effects in orthotopic HCC mouse models.
• Combination therapy with sorafenib showed improved therapeutic outcomes, highlighting the system's potential for synergistic treatment.

Conclusions:

• The Hep@PGEA/pCas9 system represents a promising nanocarrier for targeted gene therapy delivery in HCC treatment.
• The system's liver-specific accumulation and enhanced therapeutic efficacy, especially in combination therapy, suggest broad potential for treating liver diseases.