2. Academic Validation
  3. Codelivery of Cas9 mRNA and guide RNAs edits hepatitis B virus episomal and integration DNA in mouse and tree shrew models

Codelivery of Cas9 mRNA and guide RNAs edits hepatitis B virus episomal and integration DNA in mouse and tree shrew models

  • Antiviral Res. 2023 May 2;105618. doi: 10.1016/j.antiviral.2023.105618.
Junzhu Yi 1,  Xinlin Lei 2,  Fangteng Guo 1,  Qiubing Chen 2,  Xueyong Chen 3,  Kaitao Zhao 1,  Chengliang Zhu 4,  Xiaoming Cheng 5,  Jiangwei Lin 6,  Hao Yin 7,  Yuchen Xia 8
Affiliations

Affiliations

  • 1 State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Hubei Jiangxia Laboratory, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan, China; TaiKang Center for Life and Medical Sciences, Wuhan University, Wuhan, China.
  • 2 TaiKang Center for Life and Medical Sciences, Wuhan University, Wuhan, China; Department of Urology, Frontier Science Centre for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, China.
  • 3 Group of Nonhuman Primates Reproductive and Stem Cell, Kunming Institute of Zoology, Kunming, Yunnan, 650223, China.
  • 4 Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan, China.
  • 5 State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Hubei Jiangxia Laboratory, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan, China.
  • 6 Group of Nonhuman Primates Reproductive and Stem Cell, Kunming Institute of Zoology, Kunming, Yunnan, 650223, China. Electronic address: linjiangwei@mail.kiz.ac.cn.
  • 7 TaiKang Center for Life and Medical Sciences, Wuhan University, Wuhan, China; Department of Urology, Frontier Science Centre for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, China; Department of Pulmonary and Critical Care Medicine, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, China; Department of Pathology, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, China; RNA Institute, Wuhan University, Wuhan, China; Wuhan Research Centre for Infectious Diseases and Cancer, Chinese Academy of Medical Sciences, Wuhan, China. Electronic address: haoyin@whu.edu.cn.
  • 8 State Key Laboratory of Virology and Hubei Province Key Laboratory of Allergy and Immunology, Hubei Jiangxia Laboratory, Institute of Medical Virology, School of Basic Medical Sciences, Wuhan University, Wuhan, China; TaiKang Center for Life and Medical Sciences, Wuhan University, Wuhan, China. Electronic address: yuchenxia@whu.edu.cn.
PMID: 37142191 DOI: 10.1016/j.antiviral.2023.105618
Abstract

With 296 million chronically infected individuals worldwide, hepatitis B virus (HBV) causes a major health burden. The major challenge to cure HBV Infection lies in the fact that the source of persistence Infection, viral episomal covalently closed circular DNA (cccDNA), could not be targeted. In addition, HBV DNA integration, although normally results in replication-incompetent transcripts, considered as oncogenic. Though several studies evaluated the potential of gene-editing approaches to target HBV, previous in vivo studies have been of limited relevance to authentic HBV Infection, as the models do not contain HBV cccDNA or feature a complete HBV replication cycle under competent host immune system. In this study, we evaluated the effect of in vivo codelivery of Cas9 mRNA and guide RNAs (gRNAs) by SM-102-based lipid nanoparticles (LNPs) on HBV cccDNA and integrated DNA in mouse and a higher species. CRISPR nanoparticle treatment decreased the levels of HBcAg, HBsAg and cccDNA in AAV-HBV1.04 transduced mouse liver by 53%, 73% and 64% respectively. In HBV infected tree shrews, the treatment achieved 70% reduction of viral RNA and 35% reduction of cccDNA. In HBV transgenic mouse, 90% inhibition of HBV RNA and 95% inhibition of DNA were observed. CRISPR nanoparticle treatment was well tolerated in both mouse and tree shrew, as no elevation of liver Enzymes and minimal off-target was observed. Our study demonstrated that SM-102-based CRISPR is safe and effective in targeting HBV episomal and integration DNA in vivo. The system delivered by SM-102-based LNPs may be used as a potential therapeutic strategy against HBV Infection.

Keywords

Adeno-associated virus; CRISPR/Cas9; Covalently closed circular DNA (cccDNA); Hepatitis B virus (HBV); Lipid nanoparticles (LNPs); SM-102; Tree shrew.

Figures
Products