目的:戊型肝炎病毒(HEV)是全球流行的病原体,每年导致两千万人感染,三百万例临床疾病,全世界有6万人死亡,严重危及孕妇和免疫功能低下的个体。与HEV相关的研究已被大大推迟,并且尚未开发出HEV特异性疗法。我们旨在通过高通量筛选发现有效的抗HEV药物,这些药物可以在体外和体内临床前动物研究中得到验证。并阐明潜在的抗病毒机制。
方法:使用适当的细胞和啮齿动物HEV感染模型,我们研究了宿主-HEV相互作用的关键通路,并通过靶向HEV复制酶的蛋白抑制对相应的抗病毒药物进行了临床前研究.
结果:我们通过对携带HEV复制子的人肝细胞进行无偏化合物文库筛选,发现了17种靶向HEV-HSP90相互作用的抑制剂。HSP90(iHSP90)抑制剂在体外显着抑制HEV复制,其功效超过常规抗病毒药物(IFNα和利巴韦林)。机械上,iHSP90处理从ORF1-HSP90复合物释放病毒复制酶ORF1蛋白,并触发ORF1快速泛素/蛋白酶体介导的降解,导致HEV复制被取消。此外,一项针对蒙古沙鼠HEV感染模型的临床前试验表明,这种新型抗HEV策略是安全的,高效,并能预防HEV引起的肝损伤。
结论:这项研究共同说明了宿主-HEV相互作用的关键蛋白抑制途径,并为将对HEV生命周期的新理解转化为临床上有希望的抗病毒药物铺平了道路。
未经评估:对于感染HEV的孕妇以及免疫功能低下的患者,缺乏适当的治疗选择。迫切需要开发安全的HEV特异性疗法。这项研究确定了新的抗病毒药物(HSP90的抑制剂),通过靶向病毒复制酶的降解显着降低HEV感染。此外,这些抗HEV药物在HEV啮齿动物模型中进行了验证,在临床前实验中发现这些抗HEV药物对于预防HEV诱导的肝损伤是安全有效的.我们的发现大大促进了对HEV病理生物学的理解和抗病毒发展的步伐。
Appropriate treatment options are lacking for hepatitis E virus (HEV)-infected pregnant women and immunocompromised individuals. Thus, we aimed to identify efficient anti-HEV drugs through high-throughput screening, validate them in vitro and in vivo (in a preclinical animal study), and elucidate their underlying antiviral mechanism of action.
Using appropriate cellular and rodent HEV infection models, we studied a critical pathway for host-HEV interactions and performed a preclinical study of the corresponding antivirals, which target proteostasis of the HEV replicase.
We found 17 inhibitors that target HEV-HSP90 interactions by unbiased compound library screening on human hepatocytes harboring an HEV replicon. Inhibitors of HSP90 (iHSP90) markedly suppressed HEV replication with efficacy exceeding that of conventional antivirals (IFNα and ribavirin) in vitro. Mechanistically, iHSP90 treatment released the viral replicase ORF1 protein from the ORF1-HSP90 complex and triggered rapid ubiquitin/proteasome-mediated degradation of ORF1, resulting in abrogated HEV replication. Furthermore, a preclinical trial in a Mongolian gerbil HEV infection model showed this novel anti-HEV strategy to be safe, efficient, and able to prevent HEV-induced liver damage.
In this study, we uncover a proteostatic pathway that is critical for host-HEV interactions and we provide a foundation from which to translate this new understanding of the HEV life cycle into clinically promising antivirals.
Appropriate treatment options for hepatitis E virus (HEV)-infected pregnant women and immunocompromised patients are lacking; hence, there is an urgent need for safe and effective HEV-specific therapies. This study identified new antivirals (inhibitors of HSP90) that significantly limit HEV infection by targeting the viral replicase for degradation. Moreover, these anti-HEV drugs were validated in an HEV rodent model and were found to be safe and efficient for prevention of HEV-induced liver injury in preclinical experiments. Our findings substantially promote the understanding of HEV pathobiology and pave the way for antiviral development.