Novel assay to detect integrated HBV DNA in urine of chronic hepatitis B patients

检测慢性乙型肝炎患者尿液中整合 HBV DNA 的新方法

• 批准号: 10539333
• 负责人: Selena Lin
• 金额: $ 29.88万
• 依托单位: JBS SCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-11 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10539333
• 关键词: Accounting; Acute Hepatitis; Affect; American; Antiviral Therapy; Biological Assay; Biological Markers; Biotechnology; Cells; Cessation of life; Characteristics; Chromosomes; Chronic; Chronic Hepatitis; Chronic Hepatitis B; Cirrhosis; Clinic; Clinical; DNA; DNA Integration; DNA Sequence; DNA Viruses; DNA biosynthesis; Data Analyses; Data Set; Detection; Development; Disease; Disease Management; Feasibility Studies; Genes; Genetic Transcription; Genome; Goals; HBV Cirrhosis; Hepatitis B; Hepatitis B Infection; Hepatitis B Surface Antigens; Hepatitis B Virus; Hepatitis B e Antigens; Hepatocyte; Liver; Liver diseases; Longitudinal Studies; Maps; Monitor; Patients; Persons; Phase; Plasma; Ploidies; Population; Positioning Attribute; Prevention; Preventive therapy; Preventive vaccine; Primary carcinoma of the liver cells; Primer Extension; Property; Public Health; RNA; Research; Residual state; Sampling; Science; Serum Markers; Study Subject; Surface Antigens; Testing; Thinking; Time; Treatment Efficacy; Urine; Validation; Viral; Virus Diseases; Virus Integration; Virus Replication; Visit; anti-hepatitis B; commercialization; comparative; computational pipelines; design; detection assay; end stage liver disease; improved; in silico; liver biopsy; liver cancer prevention; next generation sequence data; novel; nucleoside analog; phase 2 study; programs; prototype; research clinical testing; sample collection; tool; treatment response; viral DNA; viral genomics; virus genetics

Novel assay to detect integrated HBV DNA in urine of chronic hepatitis B patients Hepatitis B virus (HBV) infection is a global public health concern. The CDC estimates that up to 2 million Americans are chronically infected with HBV. HBV infection causes acute and chronic hepatitis, the latter often leading to severe liver diseases such cirrhosis and HBV-associated hepatocellular carcinoma (HBV-HCC). HBV-HCC accounts for 50% of HCC cases worldwide. HBV is a DNA virus, and its partial genome can integrate into a host chromosome. Although HBV DNA replication can be suppressed effectively by nucleoside analogs, integrated viral DNA and covalently closed circular HBV DNA (cccDNA) persist in hepatocytes even after prolonged antiviral therapy. Continuous antiviral treatment is necessary to keep viral replication suppressed due to its ineffectiveness in eliminating the “remnant” HBV DNA (i.e., integrated HBV DNA and cccDNA) fully. Currently, the treatment goal is to achieve a functional cure, defined as the clearance of hepatitis B surface antigen (HBsAg), without definitively eliminating integrated or cccDNA. NO clinical test, however, is currently capable of identifying this “cured” population. The current thinking is that after prolonged effective anti-HBV therapy, integrated HBV DNA is the major species of residual HBV DNA in liver and most HBsAg in HBeAg-negative patients is derived from this DNA. The ability to detect integrated DNA in patients with chronic hepatitis B is an urgent unmet need. The goal of this application is to develop a noninvasive urine test that can detect and characterize integrated HBV DNA, and be used to monitor antiviral treatment efficacy and identify “cured” subjects who can be taken off the lifelong therapy. In the phase I feasibility study, we will improve our current prototype HBV-host junction detection assay and develop a computation pipeline, HBVJSeq, for comprehensive profiling of HBV integration and HBV genetics (Aim 1). In Aim 2, we will perform comparative analyses to correlate quantitative and qualitative characteristics of integrated DNA in urine and plasma with the levels of HBsAg and HBV pregenomic RNA (pgRNA) in hepatitis B e-antigen (HBeAg)-negative patients. Results of these analyses will be used to determine if cell-free integrated HBV DNA can be developed as a biomarker to identify “cured” patients in HBeAg-negative populations, warranting a phase II study to bring the test to the clinic for disease management.

检测慢性乙型肝炎患者尿液中整合 HBV DNA 的新方法 CDC 估计，乙型肝炎病毒 (HBV) 感染是一个全球性的公共卫生问题。 数以百万计的美国人慢性感染乙肝病毒，乙肝病毒感染可导致急性和慢性。 肝炎，后者通常会导致严重的肝脏疾病，例如肝硬化和乙型肝炎病毒相关的疾病 肝细胞癌 (HBV-HCC) 占全球 HCC 病例的 50%。 DNA病毒，其部分基因组可以整合到宿主染色体中。 核苷类似物、整合病毒DNA和共价键可以有效抑制复制 即使经过长期抗病毒治疗，闭环 HBV DNA (cccDNA) 仍然存在于肝细胞中。 持续的抗病毒治疗对于抑制病毒复制是必要的，因为 无法完全消除“残余”HBV DNA（即整合的 HBV DNA 和 cccDNA）。 目前，治疗目标是实现功能性治愈，即清除乙型肝炎 表面抗原 (HBsAg)，未明确消除整合或 cccDNA 临床测试， 然而，目前能够识别出这个“治愈”的人群。目前的想法是，之后。 长期有效的抗HBV治疗，整合的HBV DNA是残留HBV DNA的主要种类 在肝脏和大多数 HBeAg 阴性患者中的 HBsAg 都源自这种 DNA 检测能力。 慢性乙型肝炎患者的整合DNA是一个迫切未满足的目标。 该应用程序是开发一种无创尿液测试，可以检测和表征综合乙型肝炎病毒 DNA，用于监测抗病毒治疗效果并确定可以接受“治愈”的受试者 在第一阶段可行性研究中，我们将改进我们当前的原型。 HBV-宿主连接检测分析并开发计算管道 HBVJSeq，用于全面 HBV 整合和 HBV 遗传学分析（目标 1）。在目标 2 中，我们将进行比较。 分析尿液中整合 DNA 的定量和定性特征 血浆中乙型肝炎 e 抗原中的 HBsAg 和 HBV 前基因组 RNA (pgRNA) 水平 (HBeAg) 阴性患者的这些分析结果将用于确定是否进行无细胞整合。 HBV DNA 可以开发为生物标志物来识别 HBeAg 阴性的“治愈”患者 人群，需要进行 II 期研究，将测试带到诊所进行疾病管理。