Integrated Single-cell Multiomic Analysis of HIV Latency Reversal Reveals Novel Regulators of Viral Reactivation

Genomics Proteomics Bioinformatics. 2024 May 9;22(1):qzae003. doi: 10.1093/gpbjnl/qzae003.

Abstract

Despite the success of antiretroviral therapy, human immunodeficiency virus (HIV) cannot be cured because of a reservoir of latently infected cells that evades therapy. To understand the mechanisms of HIV latency, we employed an integrated single-cell RNA sequencing (scRNA-seq) and single-cell assay for transposase-accessible chromatin with sequencing (scATAC-seq) approach to simultaneously profile the transcriptomic and epigenomic characteristics of ∼ 125,000 latently infected primary CD4+ T cells after reactivation using three different latency reversing agents. Differentially expressed genes and differentially accessible motifs were used to examine transcriptional pathways and transcription factor (TF) activities across the cell population. We identified cellular transcripts and TFs whose expression/activity was correlated with viral reactivation and demonstrated that a machine learning model trained on these data was 75%-79% accurate at predicting viral reactivation. Finally, we validated the role of two candidate HIV-regulating factors, FOXP1 and GATA3, in viral transcription. These data demonstrate the power of integrated multimodal single-cell analysis to uncover novel relationships between host cell factors and HIV latency.

Keywords: HIV latency reversal; HIV-regulating factor; Machine learning; Primary CD4+ T cell; Single-cell multiomics.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • CD4-Positive T-Lymphocytes* / metabolism
  • CD4-Positive T-Lymphocytes* / virology
  • Forkhead Transcription Factors / genetics
  • Forkhead Transcription Factors / metabolism
  • GATA3 Transcription Factor* / genetics
  • GATA3 Transcription Factor* / metabolism
  • Gene Expression Regulation, Viral
  • HIV Infections / genetics
  • HIV Infections / metabolism
  • HIV Infections / virology
  • HIV-1* / genetics
  • HIV-1* / physiology
  • Humans
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Single-Cell Analysis* / methods
  • Transcriptome / genetics
  • Virus Activation* / genetics
  • Virus Latency* / genetics

Substances

  • GATA3 Transcription Factor
  • Forkhead Transcription Factors
  • GATA3 protein, human
  • Repressor Proteins