Abstract：
Persistent high-risk human papilloma virus (HR-HPV) infection is the main risk factor for cervical cancer, threatening women\'s health. Despite growing prophylactic vaccination, annual cervical cancer cases are still increasing and show a trend of younger onset age. However, therapeutic approaches towards HPV infection are still limited. 25-hydrocholesterol (25HC) has a wide-spectrum inhibitory effect on a variety of viruses. To explore efficient interventions to restrict HPV infection at an early time, we applied different pseudoviruses (PsV) to evaluate anti-HPV efficacy of 25HC. We tested PsV inhibition by 25HC in cervical epithelial-derived HeLa and C-33A cells, using high-risk (HPV16, HPV18, HPV59), possibly carcinogenic (HPV73), and low-risk (HPV6) HPV PsVs. Then we established murine genital HPV PsV infection models and applied IVIS to evaluate anti-HPV efficacy of 25HC in vivo. Next, with the help of confocal imaging, we targeted 25HC activity at filopodia upon HPV exposure. After that, we used RNA-seq and Western blot analysis to investigate (1) how 25HC disturbs actin cytoskeleton remodeling during HPV infection and (2) how prenylation regulates the cytoskeletal remodeling signaling pathway. Our findings suggest that 25HC perturbs F-actin rearrangement by reducing small GTPase prenylation. In this way, the phenomenon of HPV virion surfing was restricted, leading to failed infection.
摘要：
高危型人乳头瘤病毒(HR-HPV)持续感染是宫颈癌的主要危险因素,威胁女性的健康。尽管越来越多的预防性疫苗接种，每年的宫颈癌病例仍在增加，并呈现出发病年龄年轻化的趋势。然而,HPV感染的治疗方法仍然有限.25-氢胆固醇（25HC）对多种病毒具有广谱抑制作用。探索有效的干预措施，以早期限制HPV感染。我们应用不同的假病毒（PsV）来评估25HC的抗HPV功效。我们测试了25HC对宫颈上皮来源的HeLa和C-33A细胞的PsV抑制作用，使用高风险（HPV16，HPV18，HPV59），可能致癌（HPV73），和低风险（HPV6）HPVPSV。然后我们建立了小鼠生殖器HPVPsV感染模型，并应用IVIS评估了体内25HC的抗HPV功效。接下来,在共聚焦成像的帮助下，我们针对HPV暴露后的丝足有25HC活性。之后,我们使用RNA-seq和Westernblot分析来研究(1)25HC如何在HPV感染期间干扰肌动蛋白细胞骨架重塑,以及(2)戊烯化如何调节细胞骨架重塑信号通路.我们的发现表明，25HC通过减少小的GTP酶异戊二烯化来干扰F-肌动蛋白重排。这样,HPV病毒体冲浪的现象受到限制，导致感染失败。
