Machado Lucas de Almeida, Gomes Marcelo Ferreira da Costa, Guimarães Ana Carolina Ramos
Laboratory for Functional Genomics and Bioinformatics, Instituto Oswaldo Cruz, Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro, Brazil.
Scientific Computing Program, Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro, Brazil.
Front Microbiol. 2019 Sep 3;10:1981. doi: 10.3389/fmicb.2019.01981. eCollection 2019.
The human immunodeficiency virus type 1 (HIV-1) has several proteins of therapeutic importance, many of which are currently used as drug targets in antiretroviral therapy. Among these proteins is the integrase, which is responsible for the integration of the viral DNA into the host genome - a crucial step for HIV-1 replication. Given the importance of this protein in the replication process, three integrase inhibitors are currently used as an option for antiretroviral therapy: Raltegravir, Elvitegravir, and Dolutegravir. However, the crescent emergence of mutations that cause resistance to these drugs has become a worldwide health problem. In this study, we compared the variability of each position of the HIV-1 integrase sequence in clinical isolates of Raltegravir-treated and drug-naïve patients by calculating their Shannon entropies. A co-occurrence network was created to explore how mutations co-occur in patients treated with Raltegravir. Then, by building tridimensional models of the HIV-1 integrase intasomes, we investigated the relationship between variability, architecture, and co-occurrence. We observed that positions bearing some of the major resistance pathways are highly conserved among non-treated patients and variable among the treated ones. The residues involved in the three main resistance-related mutations could be identified in the same group when the positions were clustered according to their entropies. Analysis of the integrase architecture showed that the high-entropy residues S119, T124, and T125, are in contact with the host DNA, and their variations may have impacts in the protein-DNA recognition. The co-occurrence network revealed that the major resistance pathways N155H and Q148HR share more mutations with each other than with the Y143R pathway, this observation corroborates the fact that the N155H pathway is most commonly converted into Q148HRK than into Y143RCH pathway in patients' isolates. The network and the structure analysis also support the hypothesis that the resistance-related E138K mutation may be a mechanism to compensate for mutations in neighbor lysine residues to maintain DNA binding. The present study reveals patterns by which the HIV-1 integrase adapts during Raltegravir therapy. This information can be useful to comprehend the impacts of the drug in the enzyme, as well as help planning new therapeutic approaches.
1型人类免疫缺陷病毒(HIV-1)有几种具有治疗重要性的蛋白质,其中许多目前在抗逆转录病毒治疗中用作药物靶点。这些蛋白质中包括整合酶,它负责将病毒DNA整合到宿主基因组中,这是HIV-1复制的关键步骤。鉴于这种蛋白质在复制过程中的重要性,目前有三种整合酶抑制剂用作抗逆转录病毒治疗的选择:拉替拉韦、埃替拉韦和多替拉韦。然而,导致对这些药物产生耐药性的突变不断出现,已成为一个全球性的健康问题。在本研究中,我们通过计算香农熵,比较了接受拉替拉韦治疗的患者和未接受过治疗的患者临床分离株中HIV-1整合酶序列每个位置的变异性。创建了一个共现网络,以探索在接受拉替拉韦治疗的患者中突变是如何共同出现的。然后,通过构建HIV-1整合酶intasome的三维模型,我们研究了变异性、结构和共现之间的关系。我们观察到,一些主要耐药途径所在的位置在未治疗的患者中高度保守,而在接受治疗的患者中则具有变异性。当根据熵对位置进行聚类时,参与三个主要耐药相关突变的残基可在同一组中被识别。对整合酶结构的分析表明,高熵残基S119、T124和T125与宿主DNA接触,它们的变异可能会影响蛋白质与DNA的识别。共现网络显示,主要耐药途径N155H和Q148HR之间相互共享的突变比与Y143R途径更多,这一观察结果证实了在患者分离株中,N155H途径最常转变为Q148HRK,而不是Y143RCH途径。网络和结构分析还支持了以下假设:与耐药相关的E138K突变可能是一种机制,用于补偿相邻赖氨酸残基的突变以维持DNA结合。本研究揭示了HIV-1整合酶在拉替拉韦治疗期间的适应模式。这些信息有助于理解药物对该酶的影响,并有助于规划新的治疗方法。