在慢性乙型肝炎患者中检测和表征乙型肝炎病毒双链线性DNA衍生的共闭环形DNA
medRxiv : the preprint server for health sciences
|February 20, 2025
概括
乙型肝炎病毒双链线性DNA (dslDNA) 形成dsl-cccDNAs,在HBeAg(+) 慢性乙型肝炎患者中更常见. 这些可能会影响HBV病变和治疗反应.
科学领域:
- 肝病学 肝病学是一种肝病学.
- 病毒学 病毒学
- 分子生物学分子生物学
背景情况:
- 乙型肝炎病毒 (HBV) 复制产生双链线性DNA (dslDNA).
- dslDNA可以通过非同类末端结合 (NHEJ) 形成dslDNA衍生的共价封闭圆形DNA (dsl-cccDNA).
- 将dsl-cccDNA与真实cccDNA区分开来是依赖于在直接重复1 (DR1) 动图附近的插入/删除 (INDEL).
研究的目的:
- 确定dsl-cccDNA在慢性乙型肝炎 (CHB) 患者中的频率和特征.
- 在ccDNA的DR2-1区域内分析INDELs,以识别dsl-cccDNA.
- 为了比较HBeAg阳性和HBeAg阴性CHB患者之间的dsl-cccDNA患病率.
主要方法:
- 针对HBV的下一代测序 (NGS) 在56个肝脏活检中从未接受抗病毒治疗的CHB患者.
- 32个dsl-cccDNA阳性候选者的鉴定.
- 使用ccDNA-PCR NGS进行dsl-cccDNA确认和表征,分析DR1周围区域的INDEL (nt 1800-1840).
主要成果:
- 在所有32个样本中检测到dsl-cccDNA,INDELs聚集在DR1区域周围,定义它.
- 在HBeAg(+) (11.32%) 和HBeAg(-) (7.72%) 的CHB患者中,dsl-cccDNA在总ccDNA中的百分比显著更高 (p=0.01).
- dsl-cccDNA多样性与ccccDNA水平 (r=0.82),HBeAg(+) 状态和血清HBV DNA水平相关.
结论:
- 在HBeAg(+) CHB患者中,dsl-cccDNA更为普遍和异质,可能是由于活跃的病毒转录.
- 复制缺陷的dsl-cccDNA可能会导致免疫逃避,HBV整合和疾病发病.
- 在HBV治疗反应中dsl-cccDNA的临床意义需要进一步研究.
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