BLOCKING HSV INFECTION WITH BORTEZOMIB

 描述(申请人提供)：单纯疱疹病毒(HSV)会导致人类终身潜伏感染。它是重大疾病的罪魁祸首，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。尽管付出了很多努力，但还没有有效的单纯疱疹病毒疫苗。对阿昔洛韦等抗病毒药物的耐药性极大地导致了严重的单纯疱疹病毒病和死亡率。需要新的、有效的抗疱疹病毒药物，而病毒进入是一个非常可取的目标。HSV的进入需要宿主26S蛋白酶体的活性，以使进入的衣壳在细胞内运输到核周。蛋白酶体抑制剂阻断疱疹病毒感染的药效学尚未被研究。蛋白酶体抑制剂Bortezomib是一种临床有效的抗肿瘤药物，已获FDA批准用于治疗血液系统恶性肿瘤。我们将实施初步研究，将Bortezomib重新用作抗疱疹病毒药物。我们的具体目的是确定Bortezomib是否在体外有效地抑制HSV感染病理生理相关细胞。 并阐明其作用机制。为了实现这一目标，我们提出了以下次级目标：(A)描述Bortezomib介导的抑制HSV-1和HSV-2感染的药效学，包括测定选择性指数；(B)确定Bortezomib对HSV临床分离株和耐药株的影响；以及(C)确定Bortezomib抑制HSV感染的机制。我们将通过测量传入衣壳转运和蛋白酶体介导的宿主核结构域10(ND10)结构的破坏来评估Bortezomib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技术。完成这项初步研究将为重新利用抗癌药物治疗单纯疱疹病毒感染奠定关键基础。

 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.