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）感染在全球范围内的患病率为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，似乎是导致美国最近HCC加倍的主要病因。目前的治疗包括聚乙二醇干扰素-1（PEG-IFN）和利巴韦林（RBV）。美国70%的患者感染基因型1，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，并且治疗禁忌症并不罕见。由于这些原因，需要开发毒性较小并导致较高SVR的其他疗法，作为替代疗法或替代疗法。我们通过质谱分析鉴定了与HCV编码蛋白NS 5A复合的热休克蛋白（HSP）HSP 40和HSP 70。我们通过共聚焦显微镜和免疫共沉淀证实了NS 5A/HSP的相互作用。HSP 40和HSP 70敲低均减少HCV细胞培养系统中的感染性病毒颗粒产生。用热休克蛋白合成抑制剂槲皮素和KNK 437处理在无毒浓度下减少了感染性颗粒的产生。这种对病毒产生的显著抑制与其已知的低毒性以及在先前和正在进行的临床试验中的使用相结合，促使该实验室向床边提议研究用槲皮素治疗HCV感染的患者。本研究的目的是进一步了解热休克蛋白40和70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。为了实现这一目标，我们提出了三个相互关联的具体目标：1。我们将确定在HCVDNA模型中热休克蛋白40和70在丙型肝炎病毒产生中的重要性。2.我们将确定热休克蛋白合成抑制剂对丙型肝炎病毒感染的影响，在HCVDNA模型。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，降低肝硬化和肝细胞癌的发生率。 公共卫生相关性：慢性丙型肝炎病毒（HCV）感染是导致肝硬化需要肝移植的头号病原体，也是最近美国肝细胞癌加倍的主要原因。我们已经确定了热休克蛋白（HSP）的40和70作为细胞蛋白，是重要的HCV感染。我们拟在本实验室研究HSP 40、HSP 70及HSP合成抑制剂对HCV感染的影响，并提出可能导致慢性HCV感染的治疗，从而降低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. PUBLIC HEALTH RELEVANCE: Chronic hepatitis C viral (HCV) infection is the number one causative agent of cirrhosis necessitating liver transplantation and is the major cause of the recent doubling of hepatocellular carcinoma in the United States. We have identified heat shock proteins (HSP)s 40 and 70 as cellular proteins that are important for HCV infection. We propose to study HSP40, HSP70 and HSP synthesis inhibitors effect on HCV infection in this bench to beside proposal that could lead to treatment of chronic HCV infection thereby reducing the incidence of cirrhosis and hepatocellular carcinoma.