非侵入性嵌合DNA谱分析可鉴定导致肝癌中病毒抗原表达的肿

StephenW

Noninvasive chimeric DNA profiling identifies tumor-originated HBV integrants contributing to viral antigen expression in liver cancer

    Wei Chen, Ke Zhang, Peiling Dong, Gregory Fanning, Chengcheng Tao, Haikun Zhang, Shicheng Guo, Zheng Wang, Yaqiang Hong, Xiaobo Yang, Shujuan Lai, Huiguo Ding, Haitao Zhao, Changqing Zeng, Ulrike Protzer & Dake Zhang

Hepatology International volume 14, pages326–337(2020)Cite this article

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Abstract
Background

Host genome integration of HBV sequence is considered to be significant in HBV antigen expression and the development of hepatocellular carcinoma (HCC).
Method

We developed a probe-based capture strategy to enrich integrated HBV DNA for deep-sequencing analysis of integration sites in paired patient samples derived from tumor, liver tissue adjacent to tumor, saliva and plasma, as a platform for exploring the correlation, significance and utility of detecting integrations in these sample types.
Results

Most significantly, alpha fetoprotein levels significantly correlated to the amounts of integrations detected in tumor. Viral-host chimeric DNA fragments were successfully detected at high sequencing coverage in plasma rather than saliva samples from HCC patients, and each fragment of this type was only seen once in plasma from chronic hepatitis B patients. Almost all plasma chimeric fragments were derived from integrations in tumor rather than in adjacent liver tissues. Over 50% of them may produce viral-host chimeric transcripts according to deep RNA sequencing in paired tissue samples. Particularly, in patients with low HBV DNA level (< 250 UI/ml), the seemingly normal HBsAg titers may be explained by larger amounts of integrations detected. Meanwhile, we developed a strategy to predict integrants by pairing breakpoints for each integration event. Among four resolved viral patterns, the majority of Pattern I events (81.2%) retained the complete opening reading frame for HBV surface proteins.
Conclusion

We achieve the efficient enrichment of plasma cell-free chimeric DNA from integration site, and demonstrate that chimeric DNA profiling in plasma is a promising noninvasive approach to monitor HBV integration in liver cancer development and to determine the ability of integrated sequences to express viral proteins that can be targeted, e.g. by immunotherapies.

StephenW

非侵入性嵌合DNA谱分析可鉴定导致肝癌中病毒抗原表达的肿瘤起源的HBV整合子

    陈玮，张克，董培玲，格里高利·范宁，陶成成，张海坤，郭诗成，王铮，洪亚强，杨晓波，赖淑娟，丁惠国，赵海涛，曾长青，乌尔里克·普罗茨和张大可

《国际肝病学》第14卷，第326–337页（2020年）

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抽象
背景

HBV序列的宿主基因组整合被认为在HBV抗原表达和肝细胞癌（HCC）的发展中具有重要意义。
方法

我们开发了一种基于探针的捕获策略，以富集整合的HBV DNA，对来自肿瘤，邻近肿瘤的肝组织，唾液和血浆的成对患者样品中的整合位点进行深度测序分析，以此作为探索相关性，意义和实用性的平台检测这些样本类型中的积分。
结果

最重要的是，甲胎蛋白水平与肿瘤中检测到的整合量显着相关。在高测序覆盖率的血浆中而非在HCC患者的唾液样本中成功检测到了病毒-宿主嵌合DNA片段，并且在慢性乙型肝炎患者的血浆中仅见过这种类型的片段。几乎所有血浆嵌合片段均来自肿瘤中而不是邻近肝脏组织中的整合。根据配对组织样品中的深RNA测序，其中超过50％可能会产生病毒-宿主嵌合转录本。特别是，在HBV DNA水平低（<250 UI / ml）的患者中，看似正常的HBsAg滴度可以通过检测到的大量整合来解释。同时，我们开发了一种通过为每个积分事件配对断点来预测积分的策略。在四个已解析的病毒模式中，大多数模式I事件（81.2％）保留了HBV表面蛋白的完整开放阅读框。
结论

我们实现了从整合位点有效富集无血浆细胞的嵌合DNA的方法，并证明了在血浆中进行嵌合DNA谱分析是一种有前途的无创方法，可用于监测肝癌发展中的HBV整合和确定整合序列表达病毒蛋白的能力。可以作为目标，例如通过免疫疗法。