EASL2015:特定基因型变异基础核心启动子 与预核区影响分析HBeA

StephenW

P0605
GENOTYPE SPECIFIC VARIATION IN THE BASAL CORE PROMOTER AND PRE CORE REGIONS IMPACTS HBeAg LEVELS DURING IMMUNE CLEARANCE DISEASE
J. Bayliss1, G. Rosenberg1, A. Thompson2, A. Gaggar3, K. Kitrinos3,
M. Subramanian3, P. Marcellin4, E. Gane5, H.L. Chan6, X. Li1,
D. Colledge1, L. Yuen1, M. Littlejohn1, R. Edwards1, K. Jackson1,
S. Bonanzinga1, R. Hammond1, S. Bowden1, S. Locarnini1, P. Revill1.
1Molecular Research and Development, Victorian Infectious Diseases
Reference Laboratory, Melbourne, 2Gastroenterology, St Vincent’s
Hospital, Fitzroy, Australia; 3Gilead Sciences, Foster City, United States;
4Hˆopital Beaujon, University of Paris, Clichy, France; 5New Zealand
Liver Transplant Unit, Auckland City Hospital, Auckland, New Zealand;
6Department of Medicine and Therapeutics, Chinese University of
Hong Kong, Hong Kong, Hong Kong, China
E-mail: [email protected]
Background and Aims: Basal core promoter (BCP) and precore
(PC) mutations first emerge prior to HBeAg seroconversion and
have been associated with lower HBeAg expression, liver fibrosis
and HCC. GS-US-174-0103 is a phase 3 study evaluating tenofovir
in HBeAg-positive chronic hepatitis B (CHB). The aim of the current study was to use next generation sequencing (NGS) to evaluate the prevalence and quantitative frequency of key mutations within the BCP and PC regions and evaluate their impact on baseline virological profiles within the 103 cohort.
Methods: Illumina MiSeq full genome NGS was performed on a
subset of 94 baseline samples from patients enrolled in study
103, using a threshold for detection of 1%. The results of
NGS were analysed against baseline demographic, clinical, and
serological data including HBVDNA, HBsAg, and HBeAg levels. In
the first instance only mutations in the BCP and PC regions were
considered.
Results: Data were available for 94 patients (median: age 32 yrs,
69% male, HBVDNA 8.43 log10 IU/mL, HBeAg 3.30 log10PEIU/mL, HBsAg 4.71 log10 IU/mL, ALT 137.5 IU/mL, METAVIR fibrosis stage 3). BCP and PC variants were detectable in 34% and 15% of patients using population sequencing respectively vs. 43% and 41% using NGS (p < 0.001). Differences were seen in the genotype (Gt) distribution of mutations within the BCP and PC regions (Table 1).
The presence of mutation at A1762 and G1764 was associated with significantly lower HBeAg expression at baseline (p < 0.0001 for both). Presence of the G1896A mutant alone was not associated with HBeAg expression. HBsAg levels (p = 0.002), A1762T (p = 0.002) and T/C1858G (p = 0.02) were independently associated with HBeAg levels at baseline (model p < 0.0001; R2= 0.31). Baseline HBsAg levels were independently associated with HBVDNA (p < 0.0001), genotype (p = 0.005) and A1762T and G1764SA (p < 0.0001 for both) (model p < 0.0001; R2= 0.48).
Table 1. Selected BCP/PC mutations detected by NGS
n BCP PC
A1762T G1764A T/C1858G G1896A
All 94 34 (36%) 34 (36%) 13 (11%) 39 (41%)
Gt A 26 5 (19%) 6 (23%) 2 (8%) 0
Gt B 19 3 (16%) 3 (16%) 0 15 (79%)
Gt C 21 18 (86%) 19 (90%) 8 (38%) 9 (43%)
Gt D 28 8 (29%) 6 (21%) 1 (4%) 15 (54%)
Conclusions: Clinically important HBV variants are frequently
present in treatment naïve patients during the immune clearance
phase of CHB prior to the initiation of therapy, often below the
threshold of detection by population based Sanger sequencing.
These include mutations associated with increased risk of disease
progression. Our data suggest that molecular differences in the
HBV genome, both within and between genotypes may contribute
to variations in disease status among patients and illustrates the
power of NGS for detecting clinically important variants.

StephenW

P0605
特定基因型变异基础核心启动子
与预核区影响分析HBeAg的水平在
免疫清除病
J. Bayliss1，G Rosenberg1，A Thompson2，A Gaggar3，K. Kitrinos3，
M. Subramanian3，体育Marcellin4，E Gane5，HL Chan6，十丽1，
D. Colledge1，L. Yuen1，M. Littlejohn1，R. Edwards1，K. Jackson1，
S. Bonanzinga1，R. Hammond1，S. Bowden1，S. Locarnini1，P. Revill1。
1Molecular研究和开发，维多利亚传染病
参考实验室，墨尔本，2Gastroenterology，圣文森
医院，菲茨罗伊，澳大利亚; 3Gilead科学，福斯特市，美国;
4Hopital Beaujon，巴黎，克利希，法国的大学; 5New新西兰
肝移植单位，奥克兰市医院，奥克兰，新西兰;
内科及药物治疗，对中国大学6Department
香港，香港，香港，中国
电子信箱：[email protected]
背景和目的：基底核心启动子（BCP）和前C
（PC）的突变出现第一前HBeAg血清转换和
已与较低的表达大三阳，肝纤维化
和肝癌。 GS-US-174-0103是一个3期研究评估替诺福韦
在HBeAg阳性慢性乙型肝炎（CHB）。目前研究的目的是使用下一代测序（NGS）评价患病和BCP和PC的区域内的关键突变的定量次数评估它们对103的队列中基线病毒学轮廓的影响。
方法：Illumina的MiSeq全基因组NGS是在执行
94基线样本集的患者参加研究
103，使用阈值进行检测的1％。结果
NGS反对基线人口，临床分析，
血清学数据，包括HBVDNA，乙肝表面抗原，e抗原和水平。在
第一个实例仅在BCP和PC机区的突变是
考虑。
结果：数据是可用的94例患者（中位数为32岁岁，
69％的男性，HBVDNA 8.43日志10 IU / mL时，大三阳3.30 log10PEIU /毫升，乙肝表面抗原4.71日志10 IU / mL时，ALT 137.5 IU / mL时，METAVIR纤维化分期3）。 BCP和PC机的变体是可检测的在34％和使用人口测序15％患者分别与43％，并使用NGS（P <0.001）41％。差异主要出现在BCP的突变和PC地区（表1）的基因型（GT）的分布。
突变在A1762和G1764的存在是与在基线显著低HBeAg的表达（P <0.0001两者）相关联。在G1896A突变单独存在不与HBeAg的表达有关。 HBsAg水平（P = 0.002），A1762T（P = 0.002）和T / C1858G（P = 0.02）独立与HBeAg的水平基线（; R2 = 0.31模型P <0.0001）。基线HBsAg水平独立与HBVDNA相关（P <0.0001），基因型（P = 0.005）和A1762T和G1764SA（P <0.0001两个）（型号为P <0.0001; R2 = 0.48）。
通过检测NGS表1. BCP / PC突变
ñBCP PC
A1762T G1764A T / C1858G G1896A
所有94 34（36％）34（36％）13（11％）39（41％）
GT A 26 5（19％），6（23％）2（8％）0
GT乙19 3（16％）3（16％）0 15（79％）
亿吨碳21 18（86％）19（90％）8（38％）9（43％）
GTð28 8（29％）6（21％）1（4％）15（54％）
结论：临床上重要的HBV变异株经常
目前在初次接受治疗的患者的免疫清除过程
CHB之前开始治疗，常常低于相
检测的人口基于阈值的Sanger测序。
这些措施包括与疾病的风险增加相关的基因突变
进展。我们的数据表明，在分子的不同
HBV基因组内和基因型之间可能有助于
到患者之间的差异在疾病状态和说明
NGS的检测临床上重要的变型能力。

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HBV基因组变异的差异，和NGS的测病毒变异能力