In the search for new antiviral therapies against human immunodeficiency virus type 1 (HIV-1), several cellular targets are being investigated. Ataxia telangiectasia and Rad3-related protein (ATR) has been implicated in HIV-1 replication, namely during retroviral DNA integration. We studied the effect of the ATR inhibitor ETP-46464 on HIV-1 replication in peripheral blood mononuclear cells (PBMCs) and in the persistently HIV-1-infected cell line H61-D. After treatment with ETP-46464, a significant decrease in virus production was observed in both cell systems. Quantification of viral DNA forms in the acutely infected PBMCs suggests that inhibition could take place in the early phase of the viral life cycle before viral DNA integration. Moreover, after treatment of H61-D cells with 3’-azido-3’-deoxythymidine (AZT), which blocks new reverse transcription events, ETP-46464 decreased viral production, suggesting that inhibition of viral replication occurred in the late phase of the life cycle after viral DNA integration. A decrease in virus production after transfection of 293T cells with an HIV-1 infectious molecular clone also suggested that the effect of ETP-46464 is exerted at a post-integration step. We propose that ETP-46464 produces its inhibitory effect on HIV-1 replication by acting in both the early and late phases of the retroviral replication cycle. Thus, ATR could represent a new target for inhibition of HIV-1 replication.
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We thank Oscar Fernandez-Capetillo for providing the ATR inhibitor and for helpful suggestions, and Mayte Coiras and José Alcamí for the pCMV-Tat 101 plasmid. This work was supported by grants SAF (2010-17226) and (2016-77894-R) from MINECO (Ministerio de Economia y Competitividad, Spain) and PI 13/02269 from FIS (Fondo de Investigación en Salud del Instituto de Salud Carlos III) and in part by the RIS-RETIC grants RD12/0017/0028 and RD16CIII/0002/0005 funded by de ISCIII FEDER). Felix Docando had a Research Assistant Grant from the Programa de Empleo Juvenil of Comunidad de Madrid.
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Supplementary material 1 Fig. S1 Toxicity of ETP-46464 in uninfected and acutely infected PBMCs. Values indicate the viable cell concentration in uninfected or infected PBMCs treated with ETP-46464 at two days post-treatment (a) and in uninfected or infected PBMCs treated with ETP-46464 after 4 days of treatment (b). The mean and standard deviation of three (a) and five (b) separate experiments are shown (PPTX 416 kb)
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Docando, F., Casado, C., Pernas, M. et al. Characterizing the antiviral effect of an ATR inhibitor on human immunodeficiency virus type 1 replication. Arch Virol 165, 683–690 (2020). https://doi.org/10.1007/s00705-020-04531-8