Heat Shock Protein Synthesis Inhibitor Treatment of Hepatitis C Viral Infection

描述（由申请人提供）：丙型肝炎病毒（HCV）感染在世界范围内的患病率为3%，是发达国家肝移植治疗肝硬化的主要原因。在美国，HCV是最常见的慢性血源性感染，影响1.8%的人口，似乎是导致最近美国HCC翻倍的主要病因。目前的治疗包括聚乙二醇化干扰素-1 （PEG-IFN）和利巴韦林（RBV）。在美国，70%的患者感染基因1型，其持续病毒学应答（SVR）仅为42-46%。一般来说，所有基因型的治疗都可能伴有不良反应，治疗禁忌症并不少见。由于这些原因，有必要开发出毒性更小、SVR更高的附加疗法，作为辅助疗法或替代疗法。我们通过质谱分析鉴定了HSP的热休克蛋白HSP40和HSP70与HCV编码蛋白NS5A的复合物。我们通过共聚焦显微镜和共免疫沉淀证实了NS5A/HSP的相互作用。HSP40和HSP70的敲除均减少了HCV细胞培养系统中感染性病毒颗粒的产生。用热休克蛋白合成抑制剂槲皮素和KNK437处理可以在无毒浓度下减少感染性颗粒的产生。槲皮素对病毒产生的显著抑制，加上其已知的低毒性，以及在以前和正在进行的临床试验中的使用，促使这一试验台到床边的建议，研究用槲皮素治疗HCV感染患者。在本研究中，我们的目标是进一步了解HSP40和HSP70以及热休克蛋白合成抑制剂对HCV感染的影响，并在I期临床试验中确定槲皮素在慢性HCV感染患者中的安全性和抗病毒活性。为此，我们提出三个相互关联的具体目标：我们将在HCVcc模型中确定热休克蛋白40和70在丙型肝炎病毒产生中的重要性。2. 我们将在HCVcc模型中确定热休克蛋白合成抑制剂对丙型肝炎病毒感染的影响。我们将通过I期临床试验测试热休克蛋白合成抑制剂槲皮素对慢性丙型肝炎病毒感染患者的临床可行性。进一步了解HCV感染中的热休克蛋白和热休克蛋白合成抑制可能有助于成功治疗慢性丙型肝炎，并减少肝硬化和肝细胞癌的发病率。

DESCRIPTION (provided by applicant): Hepatitis C virus (HCV) infection has a worldwide prevalence of 3% and is the main entity responsible for liver transplantation in developed countries for treatment of cirrhosis. In the United States, HCV is the most common chronic blood borne infection affecting 1.8% of the population and appears to be the major etiologic factor responsible for the recent doubling of HCC in the United States. Current therapy consists of pegylated interferon-1 (PEG-IFN) and ribavirin (RBV). 70% of patients in the United States are infected with genotype 1 for which sustained virologic response (SVR) is only 42-46%. Generally, therapy of all genotypes can be accompanied by adverse effects and contraindications to therapy are not infrequent. For these reasons there is the need to develop additional therapies that are less toxic and result in higher SVR either as adjuncts or replacement therapies. We have identified the heat shock proteins (HSP)s HSP40 and HSP70 in complex with the HCV encoded protein NS5A through mass spectrometric analysis. We confirmed an NS5A/HSP interaction by confocal microscopy and coimmunoprecipitation. HSP40 and HSP70 knockdown both reduced infectious viral particle production in a HCV cell culture system. Treatment with the heat shock protein synthesis inhibitors Quercetin and KNK437 reduced infectious particle production at non-toxic concentrations. This striking inhibition of virus production combined with its known low toxicity and use in previous and ongoing clinical trials serves to motivate this bench to bedside proposal to study treat HCV infected patients with Quercetin. In this proposal, our goals are to further understand the impact of HSP40 and HSP70 and heat shock protein synthesis inhibitors on HCV infection and determine Quercetin's safety and antiviral activity in patients suffering from chronic HCV infection in a phase I clinical trial. To achieve this we propose three interrelated specific aims: 1. We will determine the importance of heat shock proteins 40 and 70 in hepatitis C virus production in the HCVcc model. 2. We will determine the impact of heat shock protein synthesis inhibitors on hepatitis C viral infection in the HCVcc model. We will test the clinical feasibility of the heat shock protein synthesis inhibitor Quercetin on patients with chronic hepatitis C viral infection through a phase I trial. Further understanding of heat shock proteins and heat shock protein synthesis inhibition in HCV infection may allow for successful treatment of chronic hepatitis C and reduce the incidence of cirrhosis and hepatocellular carcinoma.