Effect of HBV DNA Methylation and the Mutant 1762T/1764A on Viral Load and HCC

描述（由申请人提供）：高HBV病毒载量与亚洲成年人的疾病进展和抗病毒治疗耐药性有关；然而，这种作用在儿童和非亚洲成年人等其他人群中的作用尚未明确（这两种人群都是巴尔的摩人群中hbv感染个体的典型代表）。此外，HBV病毒复制率的病毒决定因素尚未在任何人群中得到充分阐明。我们的研究小组最近报道了两项研究结果，可以为这些决定因素提供新的线索：1)HBV DNA甲基化发生在非整合和整合HBV中，并可能下调病毒复制；2)新型双突变HBV 1762T/1764A， HBV核心启动子和HBV S基因序列重叠，增加了亚洲成年人的病毒复制。与其他HCC生物标志物相比，该突变体在诊断和预测亚洲成人肝细胞癌（HCC）方面具有更高的敏感性和特异性。我们建议在数据库和临床试验中使用这些新发现。该数据库将包含所有在约翰霍普金斯医疗机构接受治疗的HBV感染者，我们估计将包括约100名儿童和青少年以及约650名成年人，其中50%是非洲裔美国人，300人为HBV/HIV合并感染；150例为HBV/HIV/HCV三感染，50例为HCC。对于数据库方案，我们将分析血清中HBV DNA甲基化和双突变，以验证两个假设：1)在血清HBV DNA甲基化的患者中，HBV DNA病毒载量将低于未甲基化的患者；2)在HBV HCC患者中，HBV病毒载量将高于年龄和性别匹配的HBV感染对照组，HBV DNA甲基化率较低，双突变频率增加。该临床试验将是一项顺序随机对照试验，在免疫耐受受试者中进行5 - 21年（HBsAg+,HBeAG+, HBV DNA bbb10 (4)cpm），以确定通过核苷类似物（NA）和免疫调节剂降低HBV病毒载量是否会比单独（NA）改善应答。组1将接受恩替卡韦（ETV）治疗8周，随后接受聚乙二醇化干扰素+ ETV治疗40周；第二组将单独接受ETV。48周时，HBeAg损失/ HBV DNA < 200cpm将定义应答。

DESCRIPTION (provided by applicant): A high HBV viral load has been implicated in both disease progression and resistance to antiviral therapy in Asian adults; however this role has not been clarified in other populations such as children and non-Asian adults (both of which typify our Baltimore population of HBV-infected individuals.) Furthermore, viral determinants of HBV viral replication rates have not been fully elucidated in any population. Our group has recently reported two findings which could shed new light on these determinants: 1) Methylation of HBV DNA occurs in non-integrated as well as integrated HBV and may down regulate viral replication and 2) The novel double mutant HBV 1762T/1764A, with overlapping sequences for the HBV core promoter and the HBV S gene, increases viral replication in Asian adults. This mutant has greater sensitivity and specificity for both diagnosing and predicting hepatocellular carcinoma (HCC) in Asian adults compared to other HCC biomarkers. We propose to use these novel findings in both a database and a clinical trial. The database will contain all HBV-infected individuals cared for in the Johns Hopkins Medical Institutions which we estimate will contain ~ 100 children and adolescents and ~650 adults of whom - 50% are African American, 300 are HBV/HIV co-infected; 150 are HBV/HIV/HCV tri-infected, and 50 have HCC. For the database protocol we will analyze sera for both HBV DNA methylation and the double mutant to test two hypotheses: 1) In patients with methylated serum HBV DNA, HBV DNA viral load will be lower compared to patients with non-methylated serum HBV DNA and 2) In patients with HBV HCC, HBV viral loads will be higher, HBV DNA methylation rates lower, and the double mutant frequency increased compared to age and sex-matched HBV-infected controls. The clinical trial will be a sequential randomized controlled trial in immunotolerant subjects 5 - 21 years (HBsAg+,HBeAG+, HBV DNA >10(4)cpm), to determine if lowering the HBV viral load by a nucleoside analogue(NA) followed by an immunomodulator will improve response compared to (NA) alone. Group 1 will receive entecavir(ETV) for 8 weeks followed by pegylated interferon + ETV for 40 weeks; Group 2 will receive ETV alone. Response will be defined by HBeAg loss/ HBV DNA < 200 cpm at 48 weeks.