Wits/SAMRC Antiviral Gene Therapy Research Unit, School of Pathology, Health Sciences Faculty, University of the Witwatersrand, Johannesburg, South Africa.
Methods Mol Biol. 2020;2115:171-183. doi: 10.1007/978-1-0716-0290-4_10.
Chronic hepatitis B, a liver disease resulting from persisting hepatitis B virus (HBV) infection, remains a global health challenge despite the availability of an effective vaccine. Various preclinical studies using adeno-associated viruses (AAVs) to deliver anti-HBV RNA interference (RNAi) activators to mediate long-lasting HBV silencing show promise. Recent positive outcomes observed in clinical trials and the FDA approval of AAV-based drugs further demonstrate the potential of AAVs in antiviral therapeutic development. However, the prevalence of neutralizing antibodies against vectors based on extant AVV capsids limits the application of these vectors in human. The exciting reports on in silico designed and in vitro synthesized ancestral AAV (Anc80L65) with a potential to evade prevailing AAV neutralizing antibodies will significantly contribute to the success of these vectors in humans. Here, we describe methods for production and in vivo characterization of Anc80L65 expressing anti-HBV RNAi activators.
慢性乙型肝炎是一种由持续的乙型肝炎病毒（HBV）感染引起的肝脏疾病，尽管有有效的疫苗可用，但仍然是一个全球性的健康挑战。各种使用腺相关病毒（AAV）向肝脏递送抗 HBV RNA 干扰（RNAi）激活物以介导持久 HBV 沉默的临床前研究显示出前景。最近在临床试验中观察到的积极结果和 FDA 批准的基于 AAV 的药物进一步证明了 AAV 在抗病毒治疗开发中的潜力。然而，针对现有 AAV 衣壳的载体的中和抗体的普遍性限制了这些载体在人类中的应用。关于在计算机中设计和在体外合成具有逃避流行的 AAV 中和抗体潜力的原始 AAV（Anc80L65）的令人兴奋的报告将极大地有助于这些载体在人类中的成功。在这里，我们描述了表达抗 HBV RNAi 激活物的 Anc80L65 的生产和体内表征方法。
