title: Phosphodiesterase 8a supports HIV-1 replication in macrophages at the level of reverse transcription.
abstract: BACKGROUND
  HIV-1 infected macrophages play a key role in HIV-1 infection. Even during anti-retroviral treatment, macrophages keep producing virus due to suboptimal tissue penetration and reduced efficacy of antiretrovirals. It is therefore of major importance to understand which host factors are involved in HIV-1 replication in macrophages. Previously, we have shown that genetic polymorphisms in phosphodiesterase 8a (PDE8A) are strongly associated with HIV-1 replication in these cells. Here we analyzed the mechanism and regulation of PDE8A in HIV-1 replication in macrophages.
  
  
  RESULTS
  PDE8A mRNA expression strongly increases upon differentiation of monocytes into macrophages, which corresponds to the increased susceptibility of mature macrophages to HIV-1. In parallel, expression of microRNA miR-145-5p, predicted to target PDE8A mRNA, strongly decreased. The interaction of miR-145-5p with the 3' UTR of PDE8A mRNA could be experimentally validated, suggesting that indeed miR-145-5p can regulate PDE8A expression levels. Knockdown of PDE8A in macrophages resulted in a decrease in total HIV-1 replication and proviral DNA levels. These observations confirm that PDE8A regulates HIV-1 replication in macrophages and that this effect is mediated through early steps in the viral replication cycle.
  
  
  CONCLUSIONS
  PDE8A is highly expressed in macrophages, and its expression is regulated by miR-145-5p. Our findings strongly suggest that PDE8A supports HIV-1 replication in macrophages and that this effect is mediated at the level of reverse transcription.
authors.affiliation: Department of Experimental Immunology, Sanquin Research, Landsteiner Laboratory and Center for Infection and Immunity (CINIMA) at the Academic Medical Center of the University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Thijs
authors.initials: T
authors.lastname: Booiman
authors.affiliation: Department of Experimental Immunology, Sanquin Research, Landsteiner Laboratory and Center for Infection and Immunity (CINIMA) at the Academic Medical Center of the University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Viviana
authors.initials: V
authors.lastname: Cobos Jiménez
authors.affiliation: Department of Experimental Immunology, Sanquin Research, Landsteiner Laboratory and Center for Infection and Immunity (CINIMA) at the Academic Medical Center of the University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Karel A
authors.initials: KA
authors.lastname: van Dort
authors.affiliation: Department of Experimental Immunology, Sanquin Research, Landsteiner Laboratory and Center for Infection and Immunity (CINIMA) at the Academic Medical Center of the University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Angélique B
authors.initials: AB
authors.lastname: van 't Wout
authors.affiliation: Department of Experimental Immunology, Sanquin Research, Landsteiner Laboratory and Center for Infection and Immunity (CINIMA) at the Academic Medical Center of the University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Neeltje A
authors.initials: NA
authors.lastname: Kootstra
mesh_terms: D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D002454:Cell Differentiation; D016207:Cytokines / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000821:virology*; D035683:MicroRNAs / Q000235:genetics; D009000:Monocytes / Q000166:cytology; D048348:Reverse Transcription; D014779:Virus Replication / Q000235:genetics*
chemical_list: D016207:Cytokines; C541010:MIRN145 microRNA, human; D035683:MicroRNAs; D015105:3',5'-Cyclic-AMP Phosphodiesterases; C112770:PDE8A protein, human