Palbociclib, a selective inhibitor of cyclin-dependent kinase4/6, blocks HIV-1 reverse transcription through the control of sterile a motif and HD domain-containing protein-1 (SAMHD1) activity

Eduardo Pauls, Roger Badia, Javier Torres-Torronteras, Alba Ruiz, Marc Permanyer, Eva Riveira-Muñoz, Bonaventura Clotet, Ramón Marti, Ester Ballana, José A. Esté

Research output: Contribution to journalArticleResearchpeer-review

25 Citations (Scopus)

Abstract

©2014 Wolters Kluwer Health | Lippincott Williams & Wilkins. Background: Sterile a motif and HD domain-containing protein-1 (SAMHD1) inhibits HIV-1 reverse transcription by decreasing the pool of intracellular deoxynucleotides. SAMHD1 is controlled by cyclin-dependent kinase (CDK)-mediated phosphorylation. However, the exact mechanism of SAMHD1 regulation in primary cells is unclear. We explore the effect of palbociclib, a CDK6 inhibitor, in HIV-1 replication. Methods: Human primary monocytes were differentiated into macrophages with monocyte-colony stimulating factor and CD4<sup>+</sup> T lymphocytes stimulated with phytohaemagglutinin (PHA)/interleukin-2. Cells were treated with palbociclib and then infected with a Green fluorescent protein-expressing HIV-1 or R5 HIV-1 BaL. Viral DNA was measured by quantitative PCR and infection assessed by flow cytometry. Deoxynucleotide triphosphate (dNTP) content was determined using a polymerasebased method. Results: Pan-CDK inhibitors AT7519, roscovitine and purvalanol A reduced SAMHD1 phosphorylation. HIV-1 replication was blocked by AT7519 (66.4 ± 3.8%; n=4), roscovitine (47.3 ± 3.9%; n=4) and purvalanol A (55.7 ±15.7%; n=4) at subtoxic concentrations. Palbociclib, a potent and selective CDK6 inhibitor, blocked SAMHD1 phosphorylation, intracellular dNTP levels, HIV-1 reverse transcription and HIV-1 replication in primary macrophages and CD4<sup>+</sup> T lymphocytes. Notably, treatment of macrophages with palbociclib led to reduced CDK2 activation, measured as the phosphorylation of the T-loop at the Thr160. The antiviral effect was lost when SAMHD1 was degraded by Vpx, providing further evidence for a role of SAMHD1 in mediating the antiretroviral effect. Conclusions: Our results indicate that SAMHD1-mediated HIV-1 restriction is controlled by CDK as previously suggested but point to a preferential role for CDK2 and CDK6 as mediators of SAMHD1 activation. Our study provides a new signaling pathway susceptible for the development of new therapeutic approaches against HIV-1 infection.
Original languageEnglish
Pages (from-to)2213-2222
JournalAIDS
Volume28
Issue number15
DOIs
Publication statusPublished - 1 Jan 2014

Keywords

  • CDK2
  • CDK6
  • HIV-1
  • Restriction factor
  • SAMHD1

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