title: An In-Depth Comparison of Latency-Reversing Agent Combinations in Various In Vitro and Ex Vivo HIV-1 Latency Models Identified Bryostatin-1+JQ1 and Ingenol-B+JQ1 to Potently Reactivate Viral Gene Expression.
abstract: The persistence of latently infected cells in patients under combinatory antiretroviral therapy (cART) is a major hurdle to HIV-1 eradication. Strategies to purge these reservoirs are needed and activation of viral gene expression in latently infected cells is one promising strategy. Bromodomain and Extraterminal (BET) bromodomain inhibitors (BETi) are compounds able to reactivate latent proviruses in a positive transcription elongation factor b (P-TEFb)-dependent manner. In this study, we tested the reactivation potential of protein kinase C (PKC) agonists (prostratin, bryostatin-1 and ingenol-B), which are known to activate NF-κB signaling pathway as well as P-TEFb, used alone or in combination with P-TEFb-releasing agents (HMBA and BETi (JQ1, I-BET, I-BET151)). Using in vitro HIV-1 post-integration latency model cell lines of T-lymphoid and myeloid lineages, we demonstrated that PKC agonists and P-TEFb-releasing agents alone acted as potent latency-reversing agents (LRAs) and that their combinations led to synergistic activation of HIV-1 expression at the viral mRNA and protein levels. Mechanistically, combined treatments led to higher activations of P-TEFb and NF-κB than the corresponding individual drug treatments. Importantly, we observed in ex vivo cultures of CD8+-depleted PBMCs from 35 cART-treated HIV-1+ aviremic patients that the percentage of reactivated cultures following combinatory bryostatin-1+JQ1 treatment was identical to the percentage observed with anti-CD3+anti-CD28 antibodies positive control stimulation. Remarkably, in ex vivo cultures of resting CD4+ T cells isolated from 15 HIV-1+ cART-treated aviremic patients, the combinations bryostatin-1+JQ1 and ingenol-B+JQ1 released infectious viruses to levels similar to that obtained with the positive control stimulation. The potent effects of these two combination treatments were already detected 24 hours post-stimulation. These results constitute the first demonstration of LRA combinations exhibiting such a potent effect and represent a proof-of-concept for the co-administration of two different types of LRAs as a potential strategy to reduce the size of the latent HIV-1 reservoirs.
authors.affiliation: Service of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium; Service des Maladies Infectieuses, Université de Liège, Centre Hospitalier Universitaire (CHU) de Liège, Domaine Universitaire du Sart-Tilman, Liège, Belgium.
authors.forename: Gilles
authors.initials: G
authors.lastname: Darcis
authors.affiliation: Service of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium.
authors.forename: Anna
authors.initials: A
authors.lastname: Kula
authors.affiliation: Service of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium.
authors.forename: Sophie
authors.initials: S
authors.lastname: Bouchat
authors.affiliation: Departments of Medicine, Microbiology, and Immunology, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: Koh
authors.initials: K
authors.lastname: Fujinaga
authors.affiliation: Laboratory of Immunology, Brugmann University Hospital, Université Libre de Bruxelles (ULB), Bruxelles, Belgium.
authors.forename: Francis
authors.initials: F
authors.lastname: Corazza
authors.affiliation: Institut Universitaire de Technologie Louis Pasteur de Schiltigheim, University of Strasbourg, Schiltigheim, France.
authors.forename: Amina
authors.initials: A
authors.lastname: Ait-Ammar
authors.affiliation: Service of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium.
authors.forename: Nadège
authors.initials: N
authors.lastname: Delacourt
authors.affiliation: Service de Virologie, Université Paris-Descartes, AP-HP, Hôpital Necker-Enfants Malades, Paris, France.
authors.forename: Adeline
authors.initials: A
authors.lastname: Melard
authors.affiliation: Service des Maladies Infectieuses, CHU St-Pierre, ULB, Bruxelles, Belgium.
authors.forename: Kabamba
authors.initials: K
authors.lastname: Kabeya
authors.affiliation: Service of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium.
authors.forename: Caroline
authors.initials: C
authors.lastname: Vanhulle
authors.affiliation: Service of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium.
authors.forename: Benoit
authors.initials: B
authors.lastname: Van Driessche
authors.affiliation: Service de Génétique, Centre Hospitalier Universitaire (CHU) de Liège, Domaine Universitaire du Sart-Tilman, Liège, Belgium.
authors.forename: Jean-Stéphane
authors.initials: JS
authors.lastname: Gatot
authors.affiliation: IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire), Illkirch-Graffenstaden, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Cherrier
authors.affiliation: Kyolab, Rua Isaura Ap. Oliviera Barbosa Terini, Sao Paulo, Brazil.
authors.forename: Luiz F
authors.initials: LF
authors.lastname: Pianowski
authors.affiliation: Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Lucio
authors.initials: L
authors.lastname: Gama
authors.affiliation: Institut Universitaire de Technologie Louis Pasteur de Schiltigheim, University of Strasbourg, Schiltigheim, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Schwartz
authors.affiliation: Service of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium.
authors.forename: Jorge
authors.initials: J
authors.lastname: Vila
authors.affiliation: Service of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium.
authors.forename: Arsène
authors.initials: A
authors.lastname: Burny
authors.affiliation: Service des Maladies Infectieuses, CHU St-Pierre, ULB, Bruxelles, Belgium.
authors.forename: Nathan
authors.initials: N
authors.lastname: Clumeck
authors.affiliation: Service des Maladies Infectieuses, Université de Liège, Centre Hospitalier Universitaire (CHU) de Liège, Domaine Universitaire du Sart-Tilman, Liège, Belgium.
authors.forename: Michel
authors.initials: M
authors.lastname: Moutschen
authors.affiliation: Service des Maladies Infectieuses, CHU St-Pierre, ULB, Bruxelles, Belgium.
authors.forename: Stéphane
authors.initials: S
authors.lastname: De Wit
authors.affiliation: Departments of Medicine, Microbiology, and Immunology, University of California, San Francisco, San Francisco, California, United States of America.
authors.forename: B Matija
authors.initials: BM
authors.lastname: Peterlin
authors.affiliation: Service de Virologie, Université Paris-Descartes, AP-HP, Hôpital Necker-Enfants Malades, Paris, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Rouzioux
authors.affiliation: Institut Universitaire de Technologie Louis Pasteur de Schiltigheim, University of Strasbourg, Schiltigheim, France; Institut de Parasitologie et de Pathologie Tropicale, EA7292, University of Strasbourg, University of Strasbourg, Strasbourg, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Rohr
authors.affiliation: Service of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium.
authors.forename: Carine
authors.initials: C
authors.lastname: Van Lint
mesh_terms: D054713:Bryostatins / Q000494:pharmacology*; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000821:virology*; D004224:Diterpenes / Q000378:metabolism; D015967:Gene Expression Regulation, Viral / Q000187:drug effects*; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D042925:Positive Transcriptional Elongation Factor B / Q000378:metabolism; D014775:Virus Activation / Q000187:drug effects; D017735:Virus Latency / Q000187:drug effects
chemical_list: D054713:Bryostatins; D004224:Diterpenes; C046785:bryostatin 1; D042925:Positive Transcriptional Elongation Factor B; C008125:ingenol