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Toxicity and in vitro activity of HIV-1 latency-reversing agents in primary CNS cells

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Abstract

Despite the success of combination antiretroviral therapy (cART), HIV persists in long lived latently infected cells in the blood and tissue, and treatment is required lifelong. Recent clinical studies have trialed latency-reversing agents (LRA) as a method to eliminate latently infected cells; however, the effects of LRA on the central nervous system (CNS), a well-known site of virus persistence on cART, are unknown. In this study, we evaluated the toxicity and potency of a panel of commonly used and well-known LRA (panobinostat, romidepsin, vorinostat, chaetocin, disulfiram, hexamethylene bisacetamide [HMBA], and JQ-1) in primary fetal astrocytes (PFA) as well as monocyte-derived macrophages as a cellular model for brain perivascular macrophages. We show that most LRA are non-toxic in these cells at therapeutic concentrations. Additionally, romidepsin, JQ-1, and panobinostat were the most potent at inducing viral transcription, with greater magnitude observed in PFA. In contrast, vorinostat, chaetocin, disulfiram, and HMBA all demonstrated little or no induction of viral transcription. Together, these data suggest that some LRA could potentially activate transcription in latently infected cells in the CNS. We recommend that future trials of LRA also examine the effects of these agents on the CNS via examination of cerebrospinal fluid.

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Acknowledgments

This study was supported by the following funding bodies: the National Health and Medical Research Council (NHMRC) of Australia (APP1051093), the National Institutes of Health (NIH) (R21 MH100594), and the NIH Delaney AIDS Research Enterprise (DARE) to find a cure collaboratory (U19 AI096109) including supplemental funding from the National Institutes for Mental Health (NIMH). LRG was supported by a NHMRC Early Career Fellowship (GNT0606967). PRG is the recipient of an Australian Research Council Future Fellowship (FT120100389). SRL is an NHMRC practitioner fellow. The authors gratefully acknowledge the contribution to this work of the Victorian Operational Infrastructure Support Program received by the Burnet Institute. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Centre for Biomedical Research, Burnet Institute, 85 Commercial Rd, Melbourne, 3004, VIC, Australia

    Lachlan R. Gray, Hung On, Emma Roberts, Hao K. Lu, Michael A. Moso, Jacqueline A. Raison, Catherine Papaioannou, Wan-Jung Cheng, Anne M. Ellett, Steve L. Wesselingh, Paul R. Gorry, Sharon R. Lewin & Melissa J. Churchill

  2. Department of Infectious Diseases, Monash University, Melbourne, VIC, Australia

    Lachlan R. Gray, Hao K. Lu, Michael A. Moso, Sharon R. Lewin & Melissa J. Churchill

  3. Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, VIC, Australia

    Hao K. Lu, Michael A. Moso, Jonathan C. Jacobson, Damian F. J. Purcell & Sharon R. Lewin

  4. Department of Microbiology and Immunology, University of Melbourne, Melbourne, VIC, Australia

    Jonathan C. Jacobson, Damian F. J. Purcell & Paul R. Gorry

  5. South Australian Health and Medical Research Institute, Adelaide, South Australia, Australia

    Steve L. Wesselingh

  6. Department of Medicine, Monash University, Melbourne, VIC, Australia

    Paul R. Gorry

  7. School of Applied Sciences, College of Science, Engineering and Health, RMIT University, Melbourne, VIC, Australia

    Paul R. Gorry

  8. Department of Microbiology, Monash University, Clayton, VIC, Australia

    Melissa J. Churchill

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  1. Lachlan R. Gray
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  2. Hung On
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  3. Emma Roberts
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  4. Hao K. Lu
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Correspondence to Lachlan R. Gray or Melissa J. Churchill.

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Melissa J. Churchill and Sharon R. Lewin are equal senior authors.

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Supplementary Figure 1

Vector map of pGBFM and the generation of single-round lentiviral LTR-luciferase reporter virus. (A) Schematic of the pGBFM vector which contains a full-length HIV-1 provirus which is devoid of gag, pol, and env, but maintains the splice donor and acceptor sites, RRE element, and has a part and full HIV-1 NL4-3 LTR at the 5′ and 3′ end, respectively. The provirus also contains firefly luciferase within nef, which acts as the reporter for any LTR activity, and maintains the native polypurine tract (PPT) element to ensure efficient reverse transcription and integration. The 5′ LTR U3 element has been replaced with the CMV promoter, but following reverse transcription and integration the 5′ LTR is replaced with the 3′ LTR containing U3, R, and U5. The KpnI and XbaI sites allow for the substitution of the NL4-3 LTR with other patient-derived LTRs. (B) Single-round lentiviral LTR-luciferase reporter virus was generated by co-transfection of pGBFM, pVSV-G, pRSV-Rev, and pMDLg/pRRE into 293 T cells. Virus was harvested 48 h later and used to infect astrocytes and macrophages. (JPG 573 kb)

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Gray, L.R., On, H., Roberts, E. et al. Toxicity and in vitro activity of HIV-1 latency-reversing agents in primary CNS cells. J. Neurovirol. 22, 455–463 (2016). https://doi.org/10.1007/s13365-015-0413-4

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