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Review
. 2024 Aug 23:6:1458037.
doi: 10.3389/fgeed.2024.1458037. eCollection 2024.

In vivo liver targeted genome editing as therapeutic approach: progresses and challenges

Chiara Simoni  1   2 , Elena Barbon  1 , Andrés F Muro  3 , Alessio Cantore  1   2
Affiliations
Review

In vivo liver targeted genome editing as therapeutic approach: progresses and challenges

Chiara Simoni et al. Front Genome Ed. .

Abstract

The liver is an essential organ of the body that performs several vital functions, including the metabolism of biomolecules, foreign substances, and toxins, and the production of plasma proteins, such as coagulation factors. There are hundreds of genetic disorders affecting liver functions and, for many of them, the only curative option is orthotopic liver transplantation, which nevertheless entails many risks and long-term complications. Some peculiar features of the liver, such as its large blood flow supply and the tolerogenic immune environment, make it an attractive target for in vivo gene therapy approaches. In recent years, several genome-editing tools mainly based on the clustered regularly interspaced short palindromic repeats associated protein 9 (CRISPR-Cas9) system have been successfully exploited in the context of liver-directed preclinical or clinical therapeutic applications. These include gene knock-out, knock-in, activation, interference, or base and prime editing approaches. Despite many achievements, important challenges still need to be addressed to broaden clinical applications, such as the optimization of the delivery methods, the improvement of the editing efficiency, and the risk of on-target or off-target unwanted effects and chromosomal rearrangements. In this review, we highlight the latest progress in the development of in vivo liver-targeted genome editing approaches for the treatment of genetic disorders. We describe the technological advancements that are currently under investigation, the challenges to overcome for clinical applicability, and the future perspectives of this technology.

Keywords: delivery methods; genome editing; lipid nano particle; liver; liver directed genome editing; viral vectors.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.

Figures

FIGURE 1
FIGURE 1
Nucleases exploited for genome editing. Schematic representation of meganuclease (A), zinc finger nuclease (ZFN) (B), transcription activator-like effector nuclease (TALEN) (C), clustered regularly interspaced short palindromic repeats associated protein 9 (CRISPR-Cas9) (D), and their respective features. Created with BioRender.com.
FIGURE 2
FIGURE 2
Timeline of in vivo liver-directed genome editing. Timeline chart highlighting crucial events regarding technological advancement and applications of in vivo liver-directed genome editing. Created with BioRender.com.
FIGURE 3
FIGURE 3
Non-homologous end joining (NHEJ)-based gene editing strategies. Scheme of NHEJ-based gene editing approaches and their possible outcomes, i.e., gene disruption via indels formation or gene insertion in the presence of a donor DNA (HITI-based or bi-directional). Created with BioRender.com.
FIGURE 4
FIGURE 4
Microhomology-mediated end joining (MMEJ)-based gene editing strategies. Scheme of MMEJ-based gene editing approaches and their possible outcomes, i.e., gene disruption via larger indels formation or gene insertion in the presence of a donor DNA (PITCh). Created with BioRender.com.
FIGURE 5
FIGURE 5
Homology-directed repair (HDR)-based gene editing strategies. Scheme of HDR-mediated gene insertion mechanisms, achieved via recombination with or without nucleases. Created with BioRender.com.
FIGURE 6
FIGURE 6
Base editing strategies. Scheme of base editing approaches and their possible outcomes, i.e., gene disruption via stop codon or alternative splice formation, or gene correction of a point mutation. Created with BioRender.com.
FIGURE 7
FIGURE 7
Prime editing strategies. Scheme of prime editing approaches and their possible outcomes, i.e., gene disruption via stop codon or alternative splice formation, gene correction, editing of up to 1 kb with two pegRNAs (PEDAR, TJ), or serine integrase-mediated long insertions (TwinPE, PASTE, PASSIGE). Created with BioRender.com.
FIGURE 8
FIGURE 8
Gene silencing or activation strategies. Scheme of gene silencing or activation mechanisms, via the combination of DNA-binding proteins with repressor or activator domains, respectively. Created with BioRender.com.
FIGURE 9
FIGURE 9
Delivery vehicles to the liver. Schematic representation of adeno-associated viral vectors (AAV) (A), adenoviruses vectors (AdV) (B), lentiviral vectors (LV) or integrase-deficient lentiviral vectors (IDLV) (C), lipid nanoparticles (LNP) (D), virus-like particles (VLP) (E), and their respective features. Created with BioRender.com.
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