CRISPR/Cas9 delivery by NIR-responsive biomimetic nanoparticles for targeted HBV therapy

Area of Science:

• Biomedical Engineering
• Gene Therapy
• Nanotechnology

Background:

• Hepatitis B virus (HBV) infection lacks curative treatments, necessitating complete elimination of the viral cccDNA.
• CRISPR/Cas9 technology offers a direct method to destroy HBV cccDNA, but efficient and safe delivery systems are lacking.
• Precise spatiotemporal control of CRISPR/Cas9 remains a significant challenge for therapeutic applications.

Purpose of the Study:

• To develop an efficient and low-immunogenicity CRISPR/Cas9 delivery system for HBV therapy.
• To investigate the efficacy of a novel nanoplatform for targeting and eliminating HBV cccDNA.
• To establish remote spatiotemporal control over CRISPR/Cas9 activity for precise genome editing.

Main Methods:

• Design and synthesis of NIR-responsive biomimetic nanoparticles (UCNPs-Cas9@CM) for Cas9 RNP delivery.
• In vitro evaluation of UCNPs-Cas9@CM in HBV-infected cells to assess inhibition of viral markers.
• In vivo studies in HBV-transgenic (HBV-Tg) mice to confirm therapeutic efficacy and safety profile.

Main Results:

• UCNPs-Cas9@CM effectively delivered Cas9 RNP, achieving significant inhibition of HBsAg, HBeAg, HBV pgRNA, HBV DNA, and cccDNA in infected cells.
• Therapeutic efficacy was confirmed in HBV-Tg mice, demonstrating substantial reduction in viral load.
• The nanoplatform exhibited minimal cytotoxicity and negligible off-target DNA damage in vivo.

Conclusions:

• UCNPs-based biomimetic nanoplatforms provide an effective CRISPR-based gene therapy approach for inhibiting HBV replication.
• This novel system shows significant potential for the efficient treatment of human HBV diseases.
• The developed nanoplatform addresses key limitations in CRISPR/Cas9 delivery for viral infections.