血清乙肝病毒的RNA衣壳前基因组RNA 可能与病毒感染的持续性

StephenW

Serum hepatitis B virus RNA is encapsidated pregenome RNA
that may be associated with persistence of viral infection
and rebound
Jie Wang1,y, Tao Shen1,y, Xiangbo Huang1,y, G. Renuka Kumar2,y, Xiangmei Chen1, Zhenzhen Zeng1,Ruiyang Zhang1, Ran Chen1, Tong Li1, Tianying Zhang3, Quan Yuan3, Pao-Chen Li2, Qi Huang2,Richard Colonno2, Jidong Jia4, Jinlin Hou5, Malcolm A. McCrae6, Zhiliang Gao7,⇑, Hong Ren8,⇑,Ningshao Xia3,⇑, Hui Zhuang
1, Fengmin Lu1,⇑
1State Key Laboratory of Natural and Biomimetic Drugs, Department of Microbiology & Infectious Disease Center, School of Basic Medical
Sciences, Peking University Health Science Center, Beijing, China;
2Assembly Biosciences, Inc., San Francisco, CA, USA;
3State Key
Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, China;
4Clinical Epidemiology and EBM Unit, Being Friendship Hospital, Capital Medical University, China;
5Hepatology Unit and
Key Laboratory for Organ Failure Research, Department of Infectious Diseases, Nanfang Hospital, Southern Medical University,
Guangzhou, China;
6The Pirbright Institute, Pirbright, UK;
7Department of Infectious Diseases, The Third Affiliated Hospital of
Sun Yat-Sen University, Guangzhou, Guangdong Province, China;
8Department of Infectious Diseases, Institute of Viral Hepatitis,
The Second Affiliated Hospital, Chongqing Medical University, Chongqing, China
Background & Aims

Hepatitis B virus (HBV) RNA in serum has recently been linked to efficacy and prognosis of chronic hepatitis B (CHB) treatment. This study explored the nature, origin, underlying mechanisms, and potential clinical significance of serum HBV RNA.
Methods

The levels of HBV DNA and RNA were determined in the supernatant of induced HepAD38, HBV-expressing HepG2.2.15 cells and primary human hepatocytes (PHH), and in the serum of transgenic mice and CHB patients. NP-40 and proteinase K treatment, sucrose density gradient centrifugation, electron microscopy, northern blot, multiple identification PCRs and 5′ rapid-amplification of cDNA ends were performed to identify the nature of serum HBV RNA.
Results

Although significantly lower than HBV DNA levels, abundant HBV RNA was present in the serum of CHB patients. A series of experiments demonstrated that serum HBV RNA was pregenome RNA (pgRNA) and present in virus-like particles. HBV pgRNA virion levels increased after blocking the reverse transcription activity of HBV DNA polymerase, and decreased after blocking the encapsidation of pgRNA. Furthermore, the presence of HBV pgRNA virion was associated with risk of viral rebound after discontinuation of nucleot(s)ide analogues (NAs) therapy in CHB patients.
Conclusions

Serum HBV RNA was confirmed to be pgRNA present in virus-like particles. HBV pgRNA virions were produced from encapsidated particles in which the pgRNA was non- or partially reverse transcribed. Clinically, HBV pgRNA virion might be a potential biomarker for monitoring safe discontinuation of NA-therapy.
Lay summary

HBV may have another virion form in which the nucleic acid is composed of RNA, not DNA. The level of HBV RNA virion in serum may be associated with risk of HBV viral rebound after withdrawal of treatment, and therefore, a potential predictive biomarker to monitor the safe discontinuation of nucleot(s)ide analogues-therapy.
Abbreviations:
HBV (hepatitis B virus), cccDNA (covalently closed circular DNA), pgRNA (pregenome RNA), NAs (nucleot(s)ide analogues), CHB (chronic hepatitis B), HCC (hepatocellular carcinoma), ETV (entecavir), LMV (lamivudine), EoT (end of treatment), ADV (adefovir dipivoxil), LdT (telbivudine)
Keywords:
Hepatitis B virus, HBV pgRNA virion, Nucleot(s)ide analogues therapy

StephenW

血清乙肝病毒的RNA衣壳前基因组RNA
可能与病毒感染的持续性有关
反弹
杰Wang1，Y，陶Shen1，Y，湘波Huang1，Y，G雷努卡Kumar2，Y，香梅臣1，真真Zeng1，瑞阳Zhang1，然臣1，佟丽1，天鹰Zhang3，泉Yuan3，侦Li2的，齐Huang2理查德Colonno2，冀东Jia4，吉林Hou5，马尔科姆A. McCrae6，志良Gao7，⇑，香港Ren8，⇑，宁绍Xia3，⇑，庄辉
1，逢珉璐，⇑
基础医学学院天然药物及仿生药物国家重点实验室，微生物学和传染病中心部，
科学，北京大学医学部，北京，中国;
2Assembly Biosciences公司，公司，旧金山，加利福尼亚，美国;
3STATE关键
分子疫苗学和分子诊断，公共卫生，厦门大学，中国学院的实验室;
4Clinical流行病学和循证医学单位，作为友谊医院，首都医科大学，中国;
5Hepatology单元和
南方医科大学器官衰竭研究部传染病，南方医院，重点实验室，
中国广州;
。第六Pirbright研究所Pirbright，英国;
传染病7Department，该第三附属医院
孙中山大学，广州市，广东省，中国;
传染病8Department，病毒性肝炎研究所
附属第二医院，重庆医科大学，重庆，中国
背景和目的

乙型肝炎病毒（HBV）在血清中的RNA最近被链接到疗效和慢性乙型肝炎（CHB）的治疗的预后。本研究探索的性质，来源，潜在的机制，以及血清HBV RNA的潜在临床意义。
方法

HBV DNA和RNA的水平在诱导HepAD38，乙肝病毒表达HepG2.2.15细胞和原代人肝细胞（PHH）中，将上清液和转基因小鼠和慢性乙型肝炎患者的血清中进行了测定。 NP-40和蛋白酶K处理，蔗糖密度梯度离心，电子显微镜，Northern印迹，多个识别的PCR和cDNA末端的5'快速扩增进行，以确定血清HBV RNA的性质。
结果

虽然比显著降低HBV DNA水平，丰富的HBV RNA存在慢性乙型肝炎患者的血清中一系列的实验证实，血清HBV RNA的是前体基因组RNA（pgRNA）和存在于病毒样颗粒。 HBV pgRNA病毒水平阻断HBV DNA聚合酶的反转录活性后增加，并且阻断pgRNA的衣壳化后降低。此外，HBV pgRNA病毒颗粒的存在下，用在慢性乙型肝炎患者nucleot（S）类似物（NAS）停药后病毒反弹的风险有关。
结论

血清HBV的RNA被证实存在于病毒样颗粒pgRNA。 HBV pgRNA病毒粒子从包被的颗粒，其中所述pgRNA是非或部分反转录产生的。在临床上，乙肝pgRNA病毒可能是用于监视NA-疗法的安全停止电势生物标志物。
莱总结

HBV可具有其中所述核酸是由RNA，而不是DNA的另一种病毒体的形式。在血清HBV RNA的病毒颗粒的水平可以与治疗的停药后的HBV病毒反弹的风险相关联，并且因此，一个潜在的预测生物标志物来监控nucleot（S）类似物疗法的安全停止。
缩写：
HBV（乙肝病毒），cccDNA的（共价闭合环状DNA），pgRNA（前基因组RNA）和NAS（nucleot（S）类似物），CHB（慢性乙型肝炎），肝癌（肝细胞癌），ETV（恩替卡韦），LMV （拉米夫定），EOT（治疗结束），ADV（阿德福韦酯），LDT（替比夫定）
关键词：
乙肝病毒，乙肝病毒pgRNA病毒粒子，Nucleot（S）类似物治疗

StephenW

http://www.journal-of-hepatology ... seases%7CHepatology

一个非常有意义的研究论文. 我不同意它的一个建议:"可能与病毒感染的持续性有关".
请注意本研究同Assembly的科学家共同合作.

MP4

鲁凤民，庄辉，北京大学
贾继东，北京友谊医院
夏宁邵，厦门大学
高志良，中山大学
任红，重庆医科大学
侯金林，南方医科大学南方医院

http://www.pirbright.ac.uk/publi ... ociated-persistence
http://www.assemblybio.com/R&D/hbv_program.cfm