Department of Pediatrics, Baylor College of Medicine, Houston, Texas, United States of America.
Summer Undergraduate Research Training Program, Baylor College of Medicine, Houston, Texas, United States of America.
PLoS One. 2020 Jul 22;15(7):e0236175. doi: 10.1371/journal.pone.0236175. eCollection 2020.
Adenoviruses cause upper respiratory infections, conjunctivitis, keratitis, and gastrointestinal illness. These can be fatal in immunocompromised individuals. Adenoviruses have also been engineered into viral vectors to deliver therapeutic genes or induce immunity as vaccine carriers. The success of ocular gene therapy is driven partly by the immunologic and biochemical influences of the intraocular environment. We have shown that versican and hyaluronan modulate adenoviral vector transgene expression through CD44 signaling. Herein we explored the role of these pathways on virus replication and viral protein expression of wild type adenovirus. We report that the addition of vitreous humor (which contains both versican and hyaluronan) increases viral hexon protein levels. Vitreous humor also increased wild type adenovirus DNA replication in vitro. Metalloproteinase and γ-secretase inhibitors, which inhibit CD44 proteolytic activation, blocked adenoviral replication in vitro. Similarly, protein kinase C and RhoA kinase inhibitors, both proteins associated with CD44 mediated pathways, also inhibited wild type adenoviral replication in vitro. Application of metalloproteinase and γ-secretase inhibitors to human conjunctival explants sharply decreased adenoviral vector gene expression. Our results demonstrate that pharmacologic delivery of these inhibitors is easily achievable. The inhibition of these enzymes should be explored as potential therapies of wild type adenoviral infections.
腺病毒可引起上呼吸道感染、结膜炎、角膜炎和胃肠道疾病。在免疫功能低下的个体中，这些疾病可能是致命的。腺病毒也被设计成病毒载体，以携带治疗基因或作为疫苗载体诱导免疫。眼基因治疗的成功部分是由于眼内环境的免疫学和生化影响。我们已经表明，软骨素聚糖和透明质酸通过 CD44 信号调节腺病毒载体转基因的表达。在此，我们探讨了这些途径对野生型腺病毒病毒复制和病毒蛋白表达的作用。我们报告说，玻璃体液（含有软骨素聚糖和透明质酸）的添加增加了病毒六邻体蛋白水平。玻璃体液还增加了野生型腺病毒在体外的 DNA 复制。金属蛋白酶和 γ-分泌酶抑制剂，可抑制 CD44 的蛋白水解激活，阻断了体外腺病毒的复制。同样，蛋白激酶 C 和 RhoA 激酶抑制剂，这两种与 CD44 介导的途径相关的蛋白，也抑制了野生型腺病毒在体外的复制。金属蛋白酶和 γ-分泌酶抑制剂在人结膜外植体上的应用显著降低了腺病毒载体基因的表达。我们的结果表明，这些抑制剂的药物输送是很容易实现的。抑制这些酶应该作为野生型腺病毒感染的潜在治疗方法进行探索。
