Identifying the source of hepatitis B surface antigen in people with hepatitis B-HIV co-infection

鉴定乙型肝炎-HIV 合并感染者的乙型肝炎表面抗原来源

• 批准号: 10326630
• 负责人: ASHWIN BALAGOPAL
• 金额: $ 24.56万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-12 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10326630
• 关键词: Address; Affect; Archives; Biological Assay; Biopsy; Blood; C-terminal; CD4 Positive T Lymphocytes; Cell Count; Cessation of life; Chronic; Chronic Active Hepatitis; Chronic Hepatitis B; Circular DNA; Cirrhosis; Complementary DNA; DNA; Data; Detection; Development; Disease; Disease Progression; Foundations; Future; Genetic Transcription; Genome; Genomics; HIV; HIV-1; Hepatitis B; Hepatitis B Surface Antigens; Hepatitis B Therapy; Hepatitis B Virus; Hepatitis B e Antigens; Hepatocyte; Human Genome; Immune; In Vitro; Individual; Infection; Knowledge; Length; Liver; Liver diseases; Malignant neoplasm of liver; Maps; Messenger RNA; Open Reading Frames; Outcome; Persons; Plasma; Poly(A) Tail; Primary carcinoma of the liver cells; Production; Research; Risk; Role; Sampling; Serum; Source; Surface; T-Cell Depletion; Techniques; Time; Tissues; Transcript; Translating; United States National Institutes of Health; Vertebral column; Viral; analog; antiretroviral therapy; base; co-infection; design; detection limit; digital; env Gene Products; innovation; liver biopsy; mortality; novel; pgRNA; therapeutic target; therapy design; tool; transcriptome sequencing

Project Summary Chronic hepatitis B (CHB) affects over 250 million people worldwide, with ~1 million annual deaths due to liver disease and hepatocellular carcinoma. Up to 28% of persons living with HIV (PLWH) also have CHB. Since HIV increases liver disease progression from CHB and because liver disease is a leading cause of mortality in PLWH taking antiretroviral therapy, developing a HBV cure is imperative. Current nucleos(t)ide (NUC) therapy can control HBV replication but cannot cure CHB because it does not eradicate the stable covalently closed circular DNA (cccDNA), the template for HBV replication, from the hepatocyte. In addition, the US FDA defines HBV cure has elimination of total hepatitis B surface antigen (tHBsAg) from blood. The simplicity of this definition is belied by the complexity of the source of tHBsAg, which derives from either the cccDNA or HBV DNA that is integrated into the host genome (iDNA). Distinguishing the contribution of these two sources to HBsAg is important to target developing a cure. Further, our data using the novel techniques of droplet digital PCR (ddPCR) demonstrate that NUCs unexpectedly decrease transcription of pgRNA from cccDNA, but whether transcription of S mRNAs, the transcripts that encode for tHBsAg, is also reduced is unknown. To address these knowledge gaps, we propose to determine the contribution of cccDNA and iDNA to tHBsAg from 69 PLWH with different stages of CHB of whom 60 have paired biopsies. In a subset of these individuals, we will examine single hepatocytes to determine the proportions of hepatocytes with iDNA and cccDNA. The 69 individuals (129 biopsies since 60 have paired biopsies) in this proposal having varying stages of HBV infection including immune active CHB (HBeAg+ and HBeAg neg), inactive CHB, and occult hepatitis B. Aim 1 will use RNA seq on bulk liver tissue from 6 individuals with CHB to construct surface (S) mRNA maps, which will allow us to determine the proportion of S mRNA that originate from cccDNA versus iDNA. The latter are distinguished because iDNA will terminate in the human genome, truncating the viral sequence at its 3’ end. The maps will then be used to find major breakpoints in S mRNAs that occur with integration, allowing development of a multiplex ddPCR to study 69 bulk liver tissues and single hepatocytes from a subset of individuals. Aim 2 will interrogate liver biopsies from the 60 individuals with longitudinal biopsies during which time they were on NUCs to understand how NUCs affect these proportions. Data from Aims 1 and 2 will be correlated with plasma quantitative HBsAg and with circulating amounts of Large, Medium, and Small HBsAg. We will also determine if CD4+ T cell depletion affects the proportion of HBsAg that derives from iDNA. Our research will broadly impact the field by using novel techniques to determine the proportions of tHBsAg from iDNA or cccDNA, which will inform the rational design of therapies for HBV cure.

项目摘要 慢性肝炎B（CHB）在全球范围内影响超过2.5亿人，每年约有100万人。 疾病和肝细胞癌 艾滋病毒增加了CHB的肝病进展，因为肝病是遗嘱中的主要原因 PLWH接受抗逆转录病毒疗法 可以控制HBV复制，但无法治愈CHB，因为它不能消除稳定的共价关闭 圆形DNA（CCCDNA），来自肝细胞的HBV复制模板。 HBV治疗从血液中消除了总肝炎表面抗原（THBSAG）。 定义是由THBSAG来源的复杂性所掩盖的，THBSAG的来源源自CCCDNA或HBV 集成到宿主基因组（IDNA）中的DNA。 HBSAG旨在进一步开发一种方法。 PCR（DDPCR）表明，NUC会意外地减少CCCDNA的PGRNA的转录，但 S mRNA的转录（编码THBSAG的转录物）是否也降低。 解决知识差距，我们建议确定CCCDNA和IDNA对SUSAG的贡献 69个PLWH具有不同的CHB阶段，其中有60个成对的活检。 将检查单个肝细胞，以确定肝细胞与IDNA和CCCDNA的比例。 在具有不同阶段的HBV阶段的提案中，有69个人（自60个以来有129个活检） 感染包括免疫活性CHB（HBEAG+和HBEAG NEG），无活性CHB和隐匿性肝炎B。AIM 1 将在6个患有CHB个体的散装肝组织上使用RNA SEQ来构建表面mRNA图，其中 将允许我们确定源自CCCDNA与IDNA的S mRNA的建议 杰出的载体将在人类基因组中终止，在3'末端截断病毒序列。 然后，将发现这些地图可以找到与集成在一起的S mRNA中的断点，从而允许 从一部分 个人。 他们在NUC上了解NUC如何影响这些比例。 与血浆定量HBSAG以及循环的AMOURGE，中和小HBSAG相关。 我们还将确定CD4+ T细胞部署是否感染了来自IDNA的HBSAG的建议。 我们的Rearch将通过使用新技术来确定THBSAG的比例来影响该领域 来自IDNA或CCCDNA，这将为HBV治疗的疗法的合理设计提供信息。