Esman Anna, Salamaikina Svetlana, Kirichenko Alina, Vinokurov Michael, Fomina Darya, Sikamov Kirill, Syrkina Arina, Pokrovskaya Anastasia, Akimkin Vasily
Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Central Research Institute of Epidemiology, 111123 Moscow, Russia.
State Research Center-Burnazyan Federal Medical Biophysical Center of Federal Medical Biological Agency, 123098 Moscow, Russia.
Viruses. 2025 Mar 25;17(4):465. doi: 10.3390/v17040465.
The persistence of human immunodeficiency virus (HIV) within viral reservoirs poses significant challenges to eradication efforts. Epigenetic alterations, including DNA methylation, are potential factors influencing the latency and persistence of HIV. This study details the development and application of techniques to assess CpG methylation in the promoter regions of the and genes, which are key HIV-1 coreceptors. Using both Sanger sequencing and pyrosequencing methods, we examined 51 biological samples from 17 people living with HIV at three time points: baseline (week 0) and post-antiretroviral therapy (ART) at weeks 24 and 48. Our results revealed that promoter CpG sites were largely unmethylated, while promoter CpGs exhibited significant variability in methylation levels. Specifically, CpG 1 showed a significant decrease in methylation from week 0 to week 48, while CpG 3 displayed a significant decrease between week 0 and week 24. These differences were statistically significant when compared with non-HIV-infected controls. These findings demonstrate distinct methylation patterns between and promoters in people living with HIV over time, suggesting that epigenetic modifications may play a role in regulating the persistence of HIV-1. Our techniques provide a reliable framework for assessing gene promoter methylation and could be applied in further research on the epigenetics of HIV.
人类免疫缺陷病毒（HIV）在病毒储存库中的持续存在对根除工作构成了重大挑战。包括DNA甲基化在内的表观遗传改变是影响HIV潜伏期和持续存在的潜在因素。本研究详细介绍了评估HIV-1关键共受体 和 基因启动子区域CpG甲基化技术的开发与应用。我们使用桑格测序法和焦磷酸测序法，在三个时间点检测了17名HIV感染者的51份生物样本：基线（第0周）以及抗逆转录病毒治疗（ART）后第24周和第48周。我们的结果显示， 启动子的CpG位点大多未甲基化，而 启动子的CpG甲基化水平存在显著差异。具体而言， 基因的CpG 1从第0周到第48周甲基化显著降低，而 基因的CpG 3在第0周到第24周显著降低。与未感染HIV的对照组相比，这些差异具有统计学意义。这些发现表明，随着时间推移，HIV感染者的 和 启动子之间存在不同的甲基化模式，提示表观遗传修饰可能在调节HIV-1的持续存在中发挥作用。我们的技术为评估基因启动子甲基化提供了一个可靠的框架，可应用于HIV表观遗传学的进一步研究。
