Combined noncanonical NF-κB agonism and targeted BET bromodomain inhibition reverse HIV latency ex vivo

Latency reversal strategies for HIV cure using inhibitor of apoptosis protein (IAP) antagonists (IAPi) induce unprecedented levels of latent reservoir expression without immunotoxicity during suppressive antiretroviral therapy (ART). However, full targeting of the reservoir may require combinatorial approaches. A Jurkat latency model screen for IAPi combination partners demonstrated synergistic latency reversal with bromodomain (BD) and extraterminal domain protein inhibitors (BETi). Mechanistic investigations using CRISPR-CAS9 and single-cell RNA-Seq informed comprehensive ex vivo evaluations of IAPi plus pan-BET, bD-selective BET, or selective BET isoform targeting in CD4+ T cells from ART-suppressed donors. IAPi+BETi treatment resulted in striking induction of cell-associated HIV gag RNA, but lesser induction of fully elongated and tat-rev RNA compared with T cell activation-positive controls. IAPi+BETi resulted in HIV protein induction in bulk cultures of CD4+ T cells using an ultrasensitive p24 assay, but did not result in enhanced viral outgrowth frequency using a standard quantitative viral outgrowth assay. This study defines HIV transcriptional elongation and splicing as important barriers to latent HIV protein expression following latency reversal, delineates the roles of BET proteins and their BDs in HIV latency, and provides a rationale for exploration of IAPi+BETi in animal models of HIV latency.

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Publication Details

Journal
UNC Libraries
Published
2024-08-30
DOI
https://doi.org/10.17615/7mta-7638
Primary Topic
Protein Degradation and Inhibitors
Type
article
Field-Weighted Citation Impact
0.00

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article

Combined noncanonical NF-κB agonism and targeted BET bromodomain inhibition reverse HIV latency ex vivo

Samuel L M Raines, Brian D. Strahl, Shane D. Falcinelli, Ronald Swanstrom et al.
UNC Libraries
Protein Degradation and Inhibitors
article

Combined noncanonical NF-κB agonism and targeted BET bromodomain inhibition reverse HIV latency ex vivo

Samuel L M Raines, Brian D. Strahl, Shane D. Falcinelli, Ronald Swanstrom, David M. Margolis, Gavin C. Sampey, Robert G. Ferris, Jennifer Kirchherr, Anne‐Marie W. Turner, JoAnn Kuruc, Edward P. Browne, Caroline E. Baker, Rab K. Prinjha, Rebecca C. Furze, Nancie M. Archin, Anthony Sanchez, Jenna Read, Cynthia L. Gay, Jackson J. Peterson, Bonnie J. Howell, Katherine S. James, Brigitte Allard, David M. Irlbeck, Inmaculada Rioja, Simon Ghofrani, Cameron Sutton, Lindsey Ingerman James, Paul Zuck, Ann Emery, Guoxin Wu, Richard M. Dunham
article en

Abstract

Latency reversal strategies for HIV cure using inhibitor of apoptosis protein (IAP) antagonists (IAPi) induce unprecedented levels of latent reservoir expression without immunotoxicity during suppressive antiretroviral therapy (ART). However, full targeting of the reservoir may require combinatorial approaches. A Jurkat latency model screen for IAPi combination partners demonstrated synergistic latency reversal with bromodomain (BD) and extraterminal domain protein inhibitors (BETi). Mechanistic investigations using CRISPR-CAS9 and single-cell RNA-Seq informed comprehensive ex vivo evaluations of IAPi plus pan-BET, bD-selective BET, or selective BET isoform targeting in CD4+ T cells from ART-suppressed donors. IAPi+BETi treatment resulted in striking induction of cell-associated HIV gag RNA, but lesser induction of fully elongated and tat-rev RNA compared with T cell activation-positive controls. IAPi+BETi resulted in HIV protein induction in bulk cultures of CD4+ T cells using an ultrasensitive p24 assay, but did not result in enhanced viral outgrowth frequency using a standard quantitative viral outgrowth assay. This study defines HIV transcriptional elongation and splicing as important barriers to latent HIV protein expression following latency reversal, delineates the roles of BET proteins and their BDs in HIV latency, and provides a rationale for exploration of IAPi+BETi in animal models of HIV latency.

UNC Libraries
University of North Carolina at Chapel Hill (US)
National Institutes of Health, Center for AIDS Research, University of Washington, Center for AIDS Research, University of North Carolina at Chapel Hill
Good health and well-being
Openalex Percentile: Top 87%
Protein Degradation and Inhibitors
DOIOpen Access PDFOpenAlex
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