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

PLoS Pathogens

Christian Schwartz¹  Francis Corazza²  Christine Rouzioux²  Luiz F. Pianowski³  Nadège Delacourt⁴  Carine Van Lint⁴  Jorge Vila⁴  Lucio Gama⁵  Amina Ait-Ammar⁶  Thomas Cherrier⁷  Olivier Rohr⁸  Kabamba Kabeya⁸  B. Matija Peterlin⁹  Michel Moutschen⁹  Caroline Vanhulle⁹  Stéphane De Wit^1,10  Anna Kula^1,10  Benoit Van Driessche^1,11  Gilles Darcis^1,11  Jean-Stéphane Gatot^1,11  Nathan Clumeck^1,11  Adeline Melard^1,11  Arsène Burny^1,11  Sophie Bouchat^1,11  Koh Fujinaga^1,11 

[1] Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America;Departments of Medicine, Microbiology, and Immunology, University of California, San Francisco, San Francisco, California, United States of America;IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire), Illkirch-Graffenstaden, France;Institut Universitaire de Technologie Louis Pasteur de Schiltigheim, University of Strasbourg, Schiltigheim, France;Kyolab, Rua Isaura Ap. Oliviera Barbosa Terini, Sao Paulo, Brazil;Laboratory of Immunology, Brugmann University Hospital, Université Libre de Bruxelles (ULB), Bruxelles, Belgium;Service de Génétique, Centre Hospitalier Universitaire (CHU) de Liège, Domaine Universitaire du Sart-Tilman, Liège, Belgium;Service de Virologie, Université Paris-Descartes, AP-HP, Hôpital Necker-Enfants Malades, Paris, France;Service des Maladies Infectieuses, CHU St-Pierre, ULB, Bruxelles, Belgium;Service des Maladies Infectieuses, Université de Liège, Centre Hospitalier Universitaire (CHU) de Liège, Domaine Universitaire du Sart-Tilman, Liège, Belgium;Service of Molecular Virology, Institut de Biologie et de Médecine Moléculaires (IBMM), Université Libre de Bruxelles (ULB), Gosselies, Belgium

关键词: HIV-1;    T cells;    Transcription factors;    Viral replication;    Drug therapy;    Cell cultures;    HIV;    Viral persistence;    latency;   

DOI  :  10.1371/journal.ppat.1005063

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

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.

【 授权许可】

CC BY