Genome-wide CRISPR screens identify combinations of candidate latency reversing agents for targeting the latent HIV-1 reservoir

Authors

Weiwei Dai, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Fengting Wu, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Natalie McMyn, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Bicna Song, Center for Genetic Medicine Research, Children's National Hospital, 111 Michigan Ave NW, Washington, DC 20010, USA.
Victoria E. Walker-Sperling, Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA.
Joseph Varriale, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Hao Zhang, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Dan H. Barouch, Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Boston, MA 02215, USA.
Janet D. Siliciano, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Wei Li, Center for Genetic Medicine Research, Children's National Hospital, 111 Michigan Ave NW, Washington, DC 20010, USA.
Robert F. Siliciano, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

Document Type

Journal Article

Publication Date

10-19-2022

Journal

Science translational medicine

Volume

14

Issue

667

DOI

10.1126/scitranslmed.abh3351

Abstract

Reversing HIV-1 latency promotes killing of infected cells and is essential for cure strategies; however, no single latency reversing agent (LRA) or LRA combination have been shown to reduce HIV-1 latent reservoir size in persons living with HIV-1 (PLWH). Here, we describe an approach to systematically identify LRA combinations to reactivate latent HIV-1 using genome-wide CRISPR screens. Screens on cells treated with suboptimal concentrations of an LRA can identify host genes whose knockout enhances viral gene expression. Therefore, inhibitors of these genes should synergize with the LRA. We tested this approach using AZD5582, an activator of the noncanonical nuclear factor κB (ncNF-κB) pathway, as an LRA and identified histone deacetylase 2 (HDAC2) and bromodomain-containing protein 2 (BRD2), part of the bromodomain and extra-terminal motif (BET) protein family targeted by BET inhibitors, as potential targets. Using CD4 T cells from PLWH, we confirmed synergy between AZD5582 and several HDAC inhibitors and between AZD5582 and the BET inhibitor, JQ1. A reciprocal screen using suboptimal concentrations of an HDAC inhibitor as an LRA identified BRD2 and ncNF-κB regulators, especially BIRC2, as synergistic candidates for use in combination with HDAC inhibition. Moreover, we identified and validated additional synergistic drug candidates in latency cell line cells and primary lymphocytes isolated from PLWH. Specifically, the knockout of genes encoding CYLD or YPEL5 displayed synergy with existing LRAs in inducing HIV mRNAs. Our study provides insights into the roles of host factors in HIV-1 reactivation and validates a system for identifying drug combinations for HIV-1 latency reversal.

APA Citation

Dai, Weiwei; Wu, Fengting; McMyn, Natalie; Song, Bicna; Walker-Sperling, Victoria E.; Varriale, Joseph; Zhang, Hao; Barouch, Dan H.; Siliciano, Janet D.; Li, Wei; and Siliciano, Robert F., "Genome-wide CRISPR screens identify combinations of candidate latency reversing agents for targeting the latent HIV-1 reservoir" (2022). GW Authored Works. Paper 1757.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/1757

Department

Genomics and Precision Medicine