Recent Progress and Future Prospective in HBV Cure by CRISPR/Cas.

Area of Science:

• Hepatology
• Molecular Biology
• Gene Therapy

Background:

• Chronic hepatitis B (CHB) is a global health concern with current treatments failing to provide a cure.
• Persistent HBV DNA (cccDNA and integrated) prevents complete eradication, leading to rebound viremia after therapy cessation.

Purpose of the Study:

• To explore CRISPR-mediated gene editing as a therapeutic strategy for achieving a functional cure for CHB.
• To evaluate CRISPR-derived base editors (BEs) as a safer alternative to CRISPR-Cas9 for HBV genome inactivation.

Main Methods:

• Investigating CRISPR-Cas9's potential and its associated risks (host genome DSBs).
• Examining CRISPR-derived base editors for precise point mutations to inactivate HBV DNA without DSBs.
• Reviewing strategies to enhance base-editing efficacy, specificity, and in vivo delivery.

Main Results:

• CRISPR-Cas9 can cleave HBV DNA but poses safety concerns due to host genome double-strand breaks.
• CRISPR-derived base editors offer a promising approach to permanently inactivate HBV by introducing stop codons without DSBs.
• Optimized base-editing strategies and non-viral delivery methods (lipid nanoparticles) show potential for clinical application.

Conclusions:

• CRISPR-mediated gene therapy, particularly using base editors, holds significant promise for a functional cure of CHB.
• Overcoming challenges in efficacy, specificity, and delivery is crucial for the clinical translation of CRISPR-based HBV therapies.