BLOCKING HSV INFECTION WITH BORTEZOMIB

 描述（由申请方提供）：单纯疱疹病毒（HSV）可导致人类终身潜伏感染。它是造成重大疾病的原因，从唇疱疹和生殖器感染到失明和致命的脑炎。该项目的长期目标是开发一种新的干预策略来治疗HSV感染。尽管做了很多努力，但没有有效的HSV疫苗。对抗病毒药物如阿昔洛韦的耐药性是导致严重HSV疾病和死亡的重要原因。需要新的、有效的抗疱疹病毒药物，并且病毒进入是高度期望的靶点。HSV进入需要宿主26 S蛋白酶体活性，以将进入的衣壳细胞内转运至核外周。尚未研究阻断疱疹病毒感染的蛋白酶体抑制剂的药效学。蛋白酶体抑制剂硼替佐米是一种临床有效的抗肿瘤药物，FDA批准用于治疗血液恶性肿瘤。我们会进行试验研究，把硼替佐米重新用作抗疱疹病毒药物。我们的具体目标是确定硼替佐米是否能有效抑制HSV感染的病理生理相关细胞在体外 并阐明其作用机制。为了实现这一点，我们提出了以下子目标：（a）描述硼替佐米介导的HSV-1和HSV-2感染抑制的药效学，包括选择性指数的测定;（B）确定硼替佐米对HSV临床分离株和耐药株的影响;（c）确定硼替佐米抑制HSV感染的机制。我们将通过测量进入的衣壳转运和蛋白酶体介导的宿主核结构域10（ND 10）结构的破坏来评估硼替佐米对病毒进入的影响。我们的实验设计采用了细胞生物学、生物化学、药理学和分子病毒学的技术。完成这项试点研究将为重新利用抗癌药物治疗HSV感染奠定关键基础。

 DESCRIPTION (provided by applicant): Herpes simplex virus (HSV) causes lifelong latent infections in humans. It is responsible for significant disease, ranging from cold sores and genital infections to blindness and fatal encephalitis. The long- term objective of this project isto develop a novel intervention strategy to treat HSV infections. Despite much effort, there is no effective HSV vaccine. Resistance to antiviral drugs, such as acyclovir, contributes significantly to serious HSV disease and mortality. New, effective anti- herpesviral drugs are needed, and virus entry is a highly desirable target. HSV entry requires host 26S proteasome activity for intracellular transport of the entering capsid to the nuclear periphery. The pharmacodynamics of proteasome inhibitors that block herpesvirus infection has not been investigated. The proteasome inhibitor bortezomib is a clinically effective anti-neoplastic drug that is FDA-approved for treatment of hematologic malignancies. We will implement pilot studies to re- purpose bortezomib as an anti-herpesviral agent. Our Specific Aim is to determine whether bortezomib is effective at inhibiting HSV infection of pathophysiologically relevant cells in vitro and to elucidate its mechanism of action. To accomplish this, we propose the following sub-aims: (a) To delineate the pharmacodynamics of bortezomib-mediated inhibition of HSV-1 and HSV-2 infection, including determination of selectivity index; (b) To determine the effect of bortezomib on clinical isolates and drug-resistant strains of HSV; and (c) To ascertain the mechanism of bortezomib inhibition of HSV infection. We will assess the effect of bortezomib on virus entry by measuring incoming capsid transport and on proteasome-mediated disruption of host nuclear domain 10 (ND10) structures. Our experimental design employs techniques of cell biology, biochemistry, pharmacology, and molecular virology. Completing this pilot study will lay the critical groundwork for re-purposing an anti-cancer drug to treat HSV infections.