HIV-1 亚型 F 整合酶催化核心结构域以外的多态性导致整合酶抑制剂耐药突变 Q148H 背景下复制能力的严重丧失。

HIV-1 subtype F integrase polymorphisms external to the catalytic core domain contribute to severe loss of replication capacity in context of the integrase inhibitor resistance mutation Q148H.

机构信息

Laboratory of Cellular Biology and Retroviruses, Unit of Virology and Molecular Epidemiology, Hospital de Pediatría "Prof. Dr. Juan P. Garrahan", Buenos Aires, Argentina.

Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.

出版信息

J Antimicrob Chemother. 2022 Sep 30;77(10):2793-2802. doi: 10.1093/jac/dkac238.

DOI:10.1093/jac/dkac238

PMID:35897124

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9989736/

Abstract

BACKGROUND

In prior studies, HIV-1 BF recombinants with subtype F integrases failed to develop resistance to raltegravir through the Q148H mutational pathway. We aimed to determine the role of subtype-specific polymorphisms in integrase on drug susceptibility, viral replication and integration.

METHODS

Integrase sequences were retrieved from the Los Alamos Database or obtained from the Garrahan HIV cohort. HIV-1 infectious molecular clones with or without Q148H (+ G140S) resistance mutations were constructed using integrases of subtype B (NL4-3) or F1(BF) ARMA159 and URTR23. Integrase chimeras were generated by reciprocal exchanges of a 200 bp fragment spanning amino acids 85-150 of the catalytic core domain (CCD) of NL4-3-Q148H and either ARMA159-Q148H or URTR23-Q148H. Viral infections were quantified by p24 ELISA and Alu-gag integration PCR assay.

RESULTS

At least 18 different polymorphisms distinguish subtype B from F1(BF) recombinant integrases. In phenotypic experiments, p24 at Day 15 post-infection was high (105-106 pg/mL) for WT and NL4-3-Q148H; by contrast, it was low (102-104 pg/mL) for both F1(BF)-Q148H + G140S viruses, and undetectable for the Q148H mutants. Compared with WT viruses, integrated DNA was reduced by 5-fold for NL4-3-Q148H (P = 0.05), 9-fold for URTR23-Q148H (P = 0.01) and 16000-fold for ARMA159-Q148H (P = 0.01). Reciprocal exchange between B and F1(BF) of an integrase CCD region failed to rescue the replicative defect of F1(BF) integrase mutants.

CONCLUSIONS

The functional impairment of Q148H in the context of subtype F integrases from BF recombinants explains the lack of selection of this pathway in vivo. Non-B polymorphisms external to the integrase CCD may influence the pathway to integrase strand transfer inhibitor resistance.

摘要

背景

在先前的研究中，带有 F 亚型整合酶的 HIV-1 BF 重组体未能通过 Q148H 突变途径发展出对拉替拉韦的耐药性。我们旨在确定整合酶中的亚型特异性多态性对药物敏感性、病毒复制和整合的作用。

方法

从洛斯阿拉莫斯数据库中检索整合酶序列，或从加拉汉艾滋病毒队列中获得。使用带有或不带有 Q148H（+G140S）耐药突变的 B 亚型（NL4-3）或 F1（BF）ARMA159 和 URTR23 的整合酶构建 HIV-1 感染性分子克隆。通过 NL4-3-Q148H 和 ARMA159-Q148H 或 URTR23-Q148H 的催化核心结构域（CCD）内的 200bp 片段的相互交换生成整合酶嵌合体。通过 p24 ELISA 和 Alu-gag 整合 PCR 测定法定量病毒感染。

结果

至少有 18 种不同的多态性将 B 亚型与 F1（BF）重组整合酶区分开来。在表型实验中，感染后第 15 天的 p24 为高（105-106pg/mL），WT 和 NL4-3-Q148H；相比之下，F1（BF）-Q148H+G140S 病毒均较低（102-104pg/mL），Q148H 突变体无法检测到。与 WT 病毒相比，NL4-3-Q148H 的整合 DNA 减少了 5 倍（P=0.05），URTR23-Q148H 减少了 9 倍（P=0.01），ARMA159-Q148H 减少了 16000 倍（P=0.01）。整合酶 CCD 区域 B 和 F1（BF）之间的相互交换未能挽救 F1（BF）整合酶突变体的复制缺陷。