利用Exosomes靶向投递CRISPR/Cas9清除乙型肝炎病毒cccDNA分子的实验研究

Chronic HBV infection remains a serious public health problem. The cccDNA that persistent in nucleus unable to achieve remove is the key obstacle for a cure of chronic hepatitis B. The strategy of specific disruption cccDNA by employing CRISPR / Cas9 bring hope to figure out the problem. However, the current research data is still very limited, It's especially inadequate in vivo that the direct evidence of CRISPR / Cas9 removed cccDNA. Moreover, efficient and safe delivery carrier for CRISPR / Cas9 is lack. The project intends to use the DHBV (Duck Hepatitis B Virus) as an experimental model, To detect directly the efficacy of CRISPR / Cas9 eliminating cccDNA, in vivo, and explore to the development of efficient and safe ways to target delivery CRISPR / Cas9 by display DHBV large envelope proteins on the surface of exosomes. Through this study, we hope to directly confirm whether CRISPR / Cas9 alone or combination with existing antiviral agents can completely eliminate cccDNA, in vivo? In addition, explore to develop of an efficient and safe carrier for CRISPR / Cas9 delivery. The study will lay the foundation for the development of novel therapeutic strategies based- CRISPR / Cas9 for HBV treatment.

慢性HBV感染仍是一个严重的公共卫生问题。无法清除细胞核内持续存在的cccDNA是慢性HBV感染难以治愈的关键障碍。利用CRISPR/Cas9特异性破坏cccDNA的新策略为解决这一难题带来了希望。但是，目前相关研究的资料还非常有限，特别是体内CRISPR/Cas9直接清除cccDNA的证据不足，同时也缺乏高效安全的CRISPR/Cas9投递载体。本项目拟使用DHBV（Duck Hepatitis B Virus）作为实验模型，直接检测CRISPR/Cas9在体内、外清除cccDNA的效能，并探索在Exosomes表面展示DHBV 大包膜蛋白靶向投递CRISPR/Cas9。通过本项目研究我们希望能直接确认CRISPR/Cas9在体内单独或与现有抗病毒药协同是否能完全清除cccDNA？并探索一种安全高效的CRISPR/Cas9投递载体，为发展基于CRISPR/Cas9的HBV治疗策略奠定基础