Targeting the hepatitis B cccDNA with a sequence-specific ARCUS nuclease to eliminate hepatitis B virus in vivo

Gorsuch, Cassandra L.; Nemec, Paige S.; Yu, Mei; Xu, Simin; Han, Dong; Smith, Jeff; Lape, Janel; Buuren, Nicholas van; Ramírez, Ricardo; Muench, Robert; Holdorf, Meghan; Feierbach, Becket; Falls, Greg; Holt, J. D.; Shoop, Wendy; Sevigny, Emma; Karriker, Forrest; Brown, Robert V.; Joshi, Amod; Goodwin, Tyler J.; Tam, Ying K.; Lin, Paulo J.C.; Semple, Sean C.; Leatherbury, Neil; Delaney, William E.; Jantz, Derek; Smith, Amy

doi:10.1016/j.ymthe.2022.05.013

Cited by 20 publications

(22 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Specific cleavage mediation of viral covalently closed circular DNA (cccDNA) via nucleases. [20] PBGENE-HBV [21] Preclinical Endonuclease I-CreI EBT107 [20] Preclinical CRISPR-Cas9…”

Section: Viral Gene Repressorsmentioning

confidence: 99%

Induction of Hepatitis B Core Protein Aggregation Targeting an Unconventional Binding Site

Khayenko,

Makbul,

Schulte

et al. 2024

Preprint

View full text Add to dashboard Cite

The hepatitis B virus (HBV) infection is a major global health problem, with chronic infection leading to liver complications and high death toll. Current treatments, such as nucleos(t)ide analogs and interferon-α, effectively suppress viral replication but rarely cure the infection. To address this, new antivirals targeting different components of the HBV molecular machinery are being developed. Here we investigated the hepatitis B core protein (HBc) that forms the viral capsids and plays a vital role in the HBV life cycle. We explored two distinct binding pockets on the HBV capsid: the central hydrophobic pocket of HBc-dimers and the pocket at the tips of capsid spikes. We synthesized a geranyl dimer that binds to the central pocket with micromolar affinity, and dimeric peptides that bind the spike-tip pocket with nanomolar affinity. Cryo-electron microscopy further confirmed the binding of peptide dimers to the capsid spike tips and their capsid-aggregating properties. Finally, we show that the peptide dimers induce HBc aggregation in vitro and in living cells. Our findings highlight two tractable sites within the HBV capsid and provide an alternative strategy to affect HBV capsids.

show abstract

“…Specific cleavage mediation of viral covalently closed circular DNA (cccDNA) via nucleases. [20] PBGENE-HBV [21] Preclinical Endonuclease I-CreI EBT107 [20] Preclinical CRISPR-Cas9…”

Section: Viral Gene Repressorsmentioning

confidence: 99%

Induction of Hepatitis B Core Protein Aggregation Targeting an Unconventional Binding Site

Khayenko,

Makbul,

Schulte

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

“…Because HBsAg secretion in chronic HBV infection largely originates from HBV DNA integrated in the host genome, mutagenizing the highly conserved domains in the S gene could potentially suppress the expression of HBsAg. In this regard, cytosine base editors have demonstrated their efficacy [ 34 , 35 ].…”

Section: Gene Editing For Hepatitis Bmentioning

confidence: 99%

Prospects for Controlling Hepatitis B Globally

Soriano,

Moreno-Torres,

Treviño

et al. 2024

Pathogens

View full text Add to dashboard Cite

Infection with the hepatitis B virus (HBV) is highly prevalent globally. Over 250 million people suffer from chronic hepatitis B, and more than 800,000 patients die each year due to hepatitis B complications, including liver cancer. Although protective HBV vaccines are recommended for all newborns, global coverage is suboptimal. In adults, sexual transmission is by far the most frequent route of contagion. The WHO estimates that 1.5 million new HBV infections occur annually. Oral nucleos(t)ide analogues entecavir and tenofovir are the most frequent antivirals prescribed as HBV therapy. Almost all patients adherent to the medication achieve undetectable plasma viremia beyond 6 months of monotherapy. However, less than 5% achieve anti-HBs seroconversion, and viral rebound occurs following drug discontinuation. Therefore, nucleos(t)ide analogues need to be lifelong. New long-acting formulations of tenofovir and entecavir are being developed that will maximize treatment benefit and overcome adherence barriers. Furthermore, new antiviral agents are in development, including entry inhibitors, capside assembly modulators, and RNA interference molecules. The use of combination therapy pursues a functional HBV cure, meaning it is negative for both circulating HBV-DNA and HBsAg. Even when this goal is achieved, the cccDNA reservoir within infected hepatocytes remains a signal of past infection, and HBV can reactivate under immune suppression. Therefore, new gene therapies, including gene editing, are eagerly being pursued to silence or definitively disrupt HBV genomes within infected hepatocytes and, in this way, ultimately cure hepatitis B. At this time, three actions can be taken to push HBV eradication globally: (1) expand universal newborn HBV vaccination; (2) perform once-in-life testing of all adults to identify susceptible HBV persons that could be vaccinated (or re-vaccinated) and unveil asymptomatic carriers that could benefit from treatment; and (3) provide earlier antiviral therapy to chronic HBV carriers, as being aviremic reduces the risk of both clinical progression and transmission.

show abstract

“…TALENs and ZFNs can efficiently disable HBV targets; however, the complexity of their production precludes more efficient screening to select the best product, and direct effect in an established pool of cccDNA in a relevant natural infection model remains to be determined. Recently, data have been reported regarding the use of a specific engineered meganuclease, ARCUS, based on a naturally occurring genome editing enzyme, I-CreI, which evolved in the algae Chlamydomonas reinhardtii, to target the HBV genome [92]. Thanks to the small size of the nuclease, clinically relevant delivery was achieved in AAV-HBV mouse and non-human primate models through the systemic administration of LNPs containing ARCUS mRNA, resulting in indel detection, significant reductions in intrahepatic AAV copy numbers in both models and durable serum HBsAg decrease in the mouse model.…”

Section: Application Of Gene Editing For the Treatment Of Diseases Ca...mentioning

confidence: 99%

“…Thanks to the small size of the nuclease, clinically relevant delivery was achieved in AAV-HBV mouse and non-human primate models through the systemic administration of LNPs containing ARCUS mRNA, resulting in indel detection, significant reductions in intrahepatic AAV copy numbers in both models and durable serum HBsAg decrease in the mouse model. Moreover, assessment of genomic insertions in host cell genomes was evaluated in primary human hepatocytes, demonstrating an inverse correlation with the specificity of the meganuclease [92].…”

Section: Application Of Gene Editing For the Treatment Of Diseases Ca...mentioning

confidence: 99%

Gene Editing Technologies to Target HBV cccDNA

Martínez

Smekalova

Combe

et al. 2022

Viruses

View full text Add to dashboard Cite

Hepatitis B virus (HBV) remains a significant cause of mortality and morbidity worldwide, since chronic HBV infection is associated with elevated risk of cirrhosis and hepatocellular carcinoma. Current licensed therapies against HBV efficiently suppress viral replication; however, they do not have significant effects on the intrahepatic covalently closed circular DNA (cccDNA) of the viral minichromosome responsible for viral persistence. Thus, life-long treatment is required to avoid viral rebound. There is a significant need for novel therapies that can reduce, silence or eradicate cccDNA, thus preventing HBV reemergence after treatment withdrawal. In this review, we discuss the latest developments and applications of gene editing and related approaches for directly targeting HBV DNA and, more specifically, cccDNA in infected hepatocytes.

show abstract

Targeting the hepatitis B cccDNA with a sequence-specific ARCUS nuclease to eliminate hepatitis B virus in vivo

Cited by 20 publications

References 38 publications

Induction of Hepatitis B Core Protein Aggregation Targeting an Unconventional Binding Site

Induction of Hepatitis B Core Protein Aggregation Targeting an Unconventional Binding Site

Prospects for Controlling Hepatitis B Globally

Gene Editing Technologies to Target HBV cccDNA

Contact Info

Product

Resources

About