Digoxin reveals a functional connection between HIV-1 integration preference and T-cell activation

PLoS Pathogens

Andy Merritt¹  Charles R. M. Bangham²  Anat Melamed²  Delphine Planas³  Petronela Ancuta³  Ian Anderson⁴  Ariberto Fassati⁴  Chen-Hsuin Lee⁴  Alexander Zhyvoloup⁴  Robin Ketteler⁵  Janos Kriston-Vizi⁵  Jean-Pierre Routy⁶ 

[1] Centre for Therapeutics Discovery, MRC Technology, Mill Hill, London, United Kingdom;Department of Medicine, Imperial College, St. Mary's Campus, London, United Kingdom;Department of Microbiology, Infectiology and Immunology, Faculty of Medicine, University of Montreal and the Research Centre of the CHUM, Montreal, Québec, Canada;Division of Infection & Immunity, University College London, London, United Kingdom;MRC Laboratory for Molecular Cell Biology, University College London, London, United Kingdom;McGill University Health Centre, Glen site, Montreal, Québec, Canada

关键词: HIV-1;    T cells;    Gene expression;    Viral gene expression;    Flow cytometry;    Cell metabolism;    Memory T cells;    Adenosine triphosphatase;   

DOI  :  10.1371/journal.ppat.1006460

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

HIV-1 integrates more frequently into transcribed genes, however the biological significance of HIV-1 integration targeting has remained elusive. Using a selective high-throughput chemical screen, we discovered that the cardiac glycoside digoxin inhibits wild-type HIV-1 infection more potently than HIV-1 bearing a single point mutation (N74D) in the capsid protein. We confirmed that digoxin repressed viral gene expression by targeting the cellular Na+/K+ ATPase, but this did not explain its selectivity. Parallel RNAseq and integration mapping in infected cells demonstrated that digoxin inhibited expression of genes involved in T-cell activation and cell metabolism. Analysis of >400,000 unique integration sites showed that WT virus integrated more frequently than N74D mutant within or near genes susceptible to repression by digoxin and involved in T-cell activation and cell metabolism. Two main gene networks down-regulated by the drug were CD40L and CD38. Blocking CD40L by neutralizing antibodies selectively inhibited WT virus infection, phenocopying digoxin. Thus the selectivity of digoxin depends on a combination of integration targeting and repression of specific gene networks. The drug unmasked a functional connection between HIV-1 integration and T-cell activation. Our results suggest that HIV-1 evolved integration site selection to couple its early gene expression with the status of target CD4+ T-cells, which may affect latency and viral reactivation.

【 授权许可】

CC BY