Viral evolution in the cell‐associated HIV‐1 DNA during early ART can lead to drug resistance and virological failure in children

Abstract

Using cell‐associated DNA and cell‐free RNA of human immunodeficiency virus type‐1 (HIV‐1), we investigated the role of drug‐resistant viral variants that emerged during early antiretroviral therapy (ART) in determining virological outcome. This case‐control study compared virologic nonresponder children (two viral loads [VLs] ≥ 200 copies/mL within 2 years of ART) and responder children (two VLs < 200 copies/mL after six months of ART) infected with HIV‐1 initiated on nonnucleoside reverse‐transcriptase inhibitor (NNRTI)‐based ART. The partial reverse‐transcriptase gene of HIV‐1 in cell‐associated DNA was genotyped using next‐generation sequencing (NGS; Illumina; threshold 0.5%; at baseline and month six of ART) and in cell‐free RNA (concurrently and at virological failure; VL > 1000 copies/mL at ≥ 12 months of ART) using the Sanger method. Among 30 nonresponders and 37 responders, baseline differences were insignificant while adherence, VL, and drug resistance mutations (DRMs) observed at month six differed significantly ( P ≥ 0.05). At month six, NGS estimated a higher number of DRMs compared with Sanger (50% vs 33%; P = 0.001). Among the nonresponders carrying a resistant virus (86.6%) at virological failure, 26% harbored clinically relevant low‐frequency DRMs in the cell‐associated DNA at month six (0.5%‐20%; K103N, G190A, Y181C, and M184I). Plasma VL of > 3 log 10copies/mL (AOR, 30.4; 95% CI, 3.3‐281; P = 0.003) and treatment‐relevant DRMs detected in the cell‐associated DNA at month six (AOR, 24.2; 95% CI, 2.6‐221; P = 0.005) were independently associated with increased risk for early virological failure. Our findings suggest that treatment‐relevant DRMs acquired in cell‐associated DNA during the first six months of ART can predict virological failure in children initiated on NNRTI‐based ART.