Targeting ABL1 or ARG Tyrosine Kinases to Restrict HIV-1 Infection in Primary CD4+ T‐Cells or in Humanized NSG Mice
| Autor: | Raymond Wong, Danila Leontyev, Donald R. Branch, Beth Binnington, Alan Cochrane, Mario A. Ostrowski, Stephen D S McCarthy, Pauline Nicoletti, Hannah N Kozlowski |
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| Rok vydání: | 2019 |
| Předmět: |
CD4-Positive T-Lymphocytes
Male Dasatinib HIV Infections 030312 virology Peripheral blood mononuclear cell Virus Mice 03 medical and health sciences Mice Inbred NOD hemic and lymphatic diseases medicine Animals Humans Pharmacology (medical) RNA Small Interfering Proto-Oncogene Proteins c-abl Protein Kinase Inhibitors 0303 health sciences Gene knockdown business.industry Protein-Tyrosine Kinases Infectious Diseases Real-time polymerase chain reaction Humanized mouse HIV-1 Cancer research Female business Tyrosine kinase Ex vivo medicine.drug |
| Zdroj: | JAIDS Journal of Acquired Immune Deficiency Syndromes. 82:407-415 |
| ISSN: | 1525-4135 |
| DOI: | 10.1097/qai.0000000000002144 |
| Popis: | Background Previous studies support dasatinib as a potent inhibitor of HIV-1 replication. However, a functional distinction between 2 kinase targets of the drug, ABL1 and ARG, has not been assessed. Setting We used primary CD4 T-cells, CD8-depleted peripheral blood mononuclear cells (PBMCs) from a treatment naive HIV-1 patient, and a humanized mouse model of HIV-1 infection. We assessed the roles of ABL1 and ARG during HIV-1 infection and use of dasatinib as a potential antiviral against HIV-1 in humanized mice. Methods Primary CD4 T-cells were administered siRNA targeting ABL1 or ARG, then infected with HIV-1 containing luciferase reporter viruses. Quantitative polymerase chain reaction of viral integration of 4 HIV-1 strains was also assessed. CD8-depleted PBMCs were treated for 3 weeks with dasatinib. NSG mice were engrafted with CD34 pluripotent stem cells from human fetal cord blood, and infected with Ba-L virus after 19 weeks. Mice were treated daily with dasatinib starting 5 weeks after infection. Results siRNA knockdown of ABL1 or ARG had no effect on viral reverse transcripts, but increased 2-LTR circles 2- to 4-fold and reduced viral integration 2- to 12-fold. siRNA knockdown of ARG increased SAMHD1 activation, whereas knockdown of either kinase reduced RNA polymerase II activation. Treating CD8-depleted PBMCs from a treatment-naive patient with 50 nM of dasatinib for 3 weeks reduced p24 levels by 99.8%. Ba-L (R5)-infected mice injected daily with dasatinib showed a 95.1% reduction in plasma viral load after 2 weeks of treatment. Conclusions We demonstrate a novel nuclear role for ABL1 and ARG in ex vivo infection experiments, and proof-of-principle use of dasatinib in a humanized mouse model of HIV-1 infection. |
| Databáze: | OpenAIRE |
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