[en] Akt signaling plays a central role in many biological processes, which are key players in human immunodeficiency virus 1 (HIV-1) pathogenesis. We found that Akt interacts with HIV-1 Nef protein. In primary T cells treated with exogenous Nef or acutely infected with Nef-expressing HIV-1 in vitro, Akt became phosphorylated on serine(473) and threonine(308). In vitro, Akt activation mediated by Nef in T-cells was blocked by HIV protease inhibitors (PI), but not by reverse transcriptase inhibitors (RTI). Ex vivo, we found that the Akt pathway is hyperactivated in peripheral blood lymphocytes (PBLs) from cART naive HIV-1-infected patients. PBLs isolated from PI-treated patients, but not from RTI-treated patients, exhibited decreased Akt activation, T-cell proliferation and IL-2 production. We found that PI but not RTI can block HIV-1 reactivation in latently infected J-Lat lymphoid cells stimulated with various stimuli. Using luciferase measurement, we further confirmed that Nef-mediated reactivation of HIV-1 from latency in 1G5 cells was blocked by PI parallel to decreased Akt activation. Our results indicate that PI-mediated blockade of Akt activation could impact the HIV-1 reservoir and support the need to further assess the therapeutic use of HIV-1 PI in order to curtail latently infected cells in HIV-1-infected patients.
Disciplines :
Genetics & genetic processes
Author, co-author :
Kumar, Amit
Abbas, Wasim
Colin, Laurence
Khan, Kashif Aziz
Bouchat, Sophie
Varin, Audrey
Larbi, Anis
GATOT, Jean-Stéphane ; Centre Hospitalier Universitaire de Liège - CHU > Service de génétique
Fayard, E., Xue, G., Parcellier, A., Bozulic, L. & Hemmings, B. A. Protein kinase B (PKB/Akt), a key mediator of the PI3K signaling pathway. Curr. Top. Microbiol. Immunol. 346, 31-56 (2010).
Malim, M. H. & Emerman, M. HIV-1 accessory proteins-ensuring viral survival in a hostile environment. Cell Host. Microbe 3, 388-398 (2008).
Maartens, G., Celum, C. & Lewin, S. R. HIV infection: epidemiology, pathogenesis, treatment, and prevention. Lancet 384, 258-271 (2014).
Deng, K. & Siliciano, R. F. HIV: Early treatment may not be early enough. Nature 512, 35-36 (2014).
Liu, X. & Kumar, A. Differential signaling mechanism for HIV-1 Nef-mediated production of IL-6 and IL-8 in human astrocytes. Sci Rep. 5, 9867 (2015).
Iordanskiy, S., Santos, S. & Bukrinsky, M. Nature, nurture and HIV: The effect of producer cell on viral physiology. Virology 443, 208-213 (2013).
Gorry, P. R. et al. Pathogenicity and immunogenicity of attenuated, nef-deleted HIV-1 strains in vivo. Retrovirology 4, 66 (2007).
Renkema, G. H. & Saksela, K. Interactions of HIV-1 NEF with cellular signal transducing proteins. Front Biosci. 5, D268-D283 (2000).
Chavez, L., Calvanese, V. & Verdin, E. HIV Latency is established directly and early in both resting and activated primary CD4 T cells. Plos Pathog. 11, e1004955 (2015).
Wolf, D. et al. HIV-1 Nef associated PAK and PI3-kinases stimulate Akt-independent Bad-phosphorylation to induce anti-apoptotic signals. Nat. Med. 7, 1217-1224 (2001).
Huse, M. The T-cell-receptor signaling network. J. Cell Sci. 122, 1269-1273 (2009).
Churchill, M. J. et al. Transcriptional activity of blood- and cerebrospinal fluid-derived nef/long-terminal repeat sequences isolated from a slow progressor infected with nef-deleted human immunodeficiency virus type 1 (HIV-1) who developed HIV-associated dementia. J. Neurovirol. 12, 219-228 (2006).
Muratori, C. et al. Massive secretion by T cells is caused by HIV Nef in infected cells and by Nef transfer to bystander cells. Cell Host. Microbe 6, 218-230 (2009).
Qiao, X. et al. Human immunodeficiency virus 1 Nef suppresses CD40-dependent immunoglobulin class switching in bystander B cells. Nat. Immunol.7, 302-310 (2006).
Diehl, N. & Schaal, H. Make yourself at home: viral hijacking of the PI3K/Akt signaling pathway. Viruses. 5, 3192-212 (2013).
Ben-Romano, R. et al. Nelfinavir-induced insulin resistance is associated with impaired plasma membrane recruitment of the PI 3-kinase effectors Akt/PKB and PKC-zeta. Diabetologia 47, 1107-17 (2004).
Srirangam, A. et al. Effects of HIV protease inhibitor ritonavir on Akt-regulated cell proliferation in breast cancer. Clin Cancer Res. 12, 1883-96 (2006).
Rajan, A. A Phase I Trial of Nelfinavir (Viracept) in Adults With Solid Tumors.2011, (Date of access:30/09/2011). Available from https://clinicaltrials.gov/ct2/show/record/NCT01445106.
Plastaras, J. P. et al. Validation and toxicity of PI3K/Akt pathway inhibition by HIV protease inhibitors in humans. Cancer Biol Ther 7, 628-35 (2008).
Blagoveshchenskaya, A. D., Thomas, L., Feliciangeli, S. F., Hung, C. H. & Thomas, G. HIV-1 Nef downregulates MHC-I by a PACS-1- and PI3K-regulated ARF6 endocytic pathway. Cell 111, 853-866 (2002).
Sarbassov, D. D., Guertin, D. A., Ali, S. M. & Sabatini, D. M. Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science 307, 1098-1101 (2005).
Lakadamyali, M., Rust, M. J. & Zhuang, X. Endocytosis of influenza viruses. Microbes Infect 6, 929-36(2004).
Henriques, C. M., Rino, J., Nibbs, R. J., Graham, G. J. & Barata, J. T. IL-7 induces rapid clathrin-mediated internalization and JAK3-dependent degradation of IL-7Ralpha in T cells. Blood. 115, 3269-77(2010).
Kotov, A., Zhou, J., Flicker, P. & Aiken, C. Association of Nef with the human immunodeficiency virus type 1 core. J Virol. 73, 8824-30 (1999).
Welker, R., Harris, M., Cardel, B. & Kräusslich, H. G. Virion incorporation of human immunodeficiency virus type 1 Nef is mediated by a bipartite membrane-targeting signal: analysis of its role in enhancement of viral infectivity. J Virol. 72, 8833-40 (1998).
Rosa, A. et al. HIV-1 Nef promotes infection by excluding SERINC5 from virion incorporation. Nature 526, 212-7 (2015).
Usami, Y., Wu, Y. & Göttlinger, H. G. SERINC3 and SERINC5 restrict HIV-1 infectivity and are counteracted by Nef. Nature 526, 218-23 (2015).
Stephens, L. et al. Protein kinase B kinases that mediate phosphatidylinositol 3,4,5-trisphosphate-dependent activation of protein kinase B. Science 279, 710-714 (1998).
Navarro, M. N. & Cantrell, D. A. Serine-threonine kinases in TCR signaling. Nat. Immunol.15, 808-814 (2014).
Parry, R. V., Riley, J. L. & Ward, S. G. Signalling to suit function: tailoring phosphoinositide 3-kinase during T-cell activation. Trends Immunol. 28, 161-168 (2007).
Wang, J. K., Kiyokawa, E., Verdin, E. & Trono, D. The Nef protein of HIV-1 associates with rafts and primes T cells for activation. Proc. Natl. Acad. Sci. USA 97, 394-399 (2000).
Fenard, D. et al. Nef is physically recruited into the immunological synapse and potentiates T cell activation early after TCR engagement. J. Immunol. 175, 6050-6057 (2005).
Thomas, R. M., Gao, L. & Wells, A. D. Signals from CD28 induce stable epigenetic modification of the IL-2 promoter. J. Immunol. 174, 4639-4646 (2005).
Gantt, S., Casper, C. & Ambinder, R. F. Insights into the broad cellular effects of nelfinavir and the HIV protease inhibitors supporting their role in cancer treatment and prevention. Curr. Opin. Oncol. 25, 495-502 (2013).
Gupta, A. K., Cerniglia, G. J., Mick, R., McKenna, W. G. & Muschel, R. J. HIV protease inhibitors block Akt signaling and radiosensitize tumor cells both in vitro and in vivo. Cancer Res. 65, 8256-8265 (2005).
Bernstein, W. B. & Dennis, P. A. Repositioning HIV protease inhibitors as cancer therapeutics. Curr Opin HIV AIDS 3, 666-75 (2008).
Wallet, M. A. et al. The HIV-1 protease inhibitor nelfinavir activates PP2 and inhibits MAPK signaling in macrophages: a pathway to reduce inflammation. J. Leukoc. Biol. 92, 795-805 (2012).
Wagner, T. A. et al. HIV latency. Proliferation of cells with HIV integrated into cancer genes contributes to persistent infection. Science 345, 570-573 (2014).
Van Lint, C., Bouchat, S. & Marcello, A. HIV-1 transcription and latency: an update. Retrovirology. 10, 67 (2013).
Hill, A. L., Rosenbloom, D. I., Fu, F., Nowak, M. A. & Siliciano, R. F. Predicting the outcomes of treatment to eradicate the latent reservoir for HIV-1. Proc. Natl. Acad. Sci. USA 111, 13475-13480 (2014).
Ho, Y. C. et al. Replication-competent noninduced proviruses in the latent reservoir increase barrier to HIV-1 cure. Cell 155, 540-551 (2013).
Deeks, S. G. et al. Towards an HIV cure: a global scientific strategy. Nat. Rev. Immunol. 12, 607-614 (2012).
Kumar, A., Abbas, W. & Herbein, G. HIV-1 latency in monocytes/macrophages. Viruses 6, 1837-1860 (2014).
Contreras, X. et al. Suberoylanilide hydroxamic acid reactivates HIV from latently infected cells. J. Biol. Chem. 284, 6782-6789 (2009).
Reuse, S. et al. Synergistic activation of HIV-1 expression by deacetylase inhibitors and prostratin: implications for treatment of latent infection. Plos. One 4, e6093 (2009).
Schrager, J. A., Der Minassian, V. & Marsh, J. W. HIV Nef increases T cell ERK MAP kinase activity. J Biol Chem. 277, 6137-42 (2002).
Xue, M. et al. HIV-1 Nef and KSHV oncogene K1 synergistically promote angiogenesis by inducing cellular miR-718 to regulate the PTEN/AKT/mTOR signaling pathway. Nucleic Acids Res. 42, 9862-79 (2014).
Neri, F., Giolo, G., Potesta, M., Petrini, S. & Doria, M. The HIV-1 Nef protein has a dual role in T cell receptor signaling in infected CD4+ T lymphocytes. Virology 410, 316-326 (2011).
James, C. O. et al. Extracellular Nef protein targets CD4+ T cells for apoptosis by interacting with CXCR4 surface receptors. J. Virol. 78, 3099-3109 (2004).
Martelli, A. M. et al. The emerging multiple roles of nuclear Akt. Biochim. Biophys. Acta 1823, 2168-2178 (2012).
Lenassi, M. et al. HIV Nef is secreted in exosomes and triggers apoptosis in bystander CD4+ T cells. Traffic 11, 110-122 (2010).
de Carvalho, J. V. et al. Nef neutralizes the ability of exosomes from CD4+ T cells to act as decoys during HIV-1 infection. Plos. One 9, e113691 (2014).
Arenaccio, C. et al. Exosomes from human immunodeficiency virus type 1 (HIV-1)-infected cells license quiescent CD4+ T lymphocytes to replicate HIV-1 through a Nef- and ADAM17-dependent mechanism. J. Virol. 88, 11529-11539 (2014).
Luo, X., Fan, Y., Park, I. W. & He, J. J. Exosomes are unlikely involved in intercellular Nef transfer. Plos. One 10, e0124436 (2015).
Deregibus, M. C. et al. HIV-1-Tat protein activates phosphatidylinositol 3-kinase/AKT-dependent survival pathways in Kaposi's sarcoma cells. J. Biol. Chem. 277, 25195-25202 (2002).
Alexander, M., Bor, Y. C., Ravichandran, K. S., Hammarskjold, M. L. & Rekosh, D. Human immunodeficiency virus type 1 Nef associates with lipid rafts to downmodulate cell surface CD4 and class I major histocompatibility complex expression and to increase viral infectivity. J. Virol. 78, 1685-1696 (2004).
Calay, D. et al. Inhibition of Akt signaling by exclusion from lipid rafts in normal and transformed epidermal keratinocytes. J. Invest Dermatol. 130, 1136-1145 (2010).
Horejsi, V. & Hrdinka, M. Membrane microdomains in immunoreceptor signaling. FEBS Lett. 588, 2392-7 (2014).
Keppler, O. T., Tibroni, N., Venzke, S., Rauch, S. & Fackler, O. T. Modulation of specific surface receptors and activation sensitization in primary resting CD4+ T lymphocytes by the Nef protein of HIV-1. J. Leukoc. Biol. 79, 616-627 (2006).
Varin, A. et al. Exogenous Nef protein activates NF-kappa B, AP-1, and c-Jun N-terminal kinase and stimulates HIV transcription in promonocytic cells. Role in AIDS pathogenesis. J. Biol. Chem. 278, 2219-2227 (2003).
Markle, T. J., Philip, M. & Brockman, M. A. HIV-1 Nef and T-cell activation: a history of contradictions. Future. Virol. 8 (2013).
Liu, X. et al. HIV-1 Nef induces CCL5 production in astrocytes through p38-MAPK and PI3K/Akt pathway and utilizes NF-kB, CEBP and AP-1 transcription factors. Sci Rep. 4, 4450 (2014).
Schrager, J. A. & Marsh, J. W. HIV-1 Nef increases T cell activation in a stimulus-dependent manner. Proc. Natl. Acad. Sci. USA 96, 8167-8172 (1999).
Park, I. W. & He, J. J. HIV-1 Nef-mediated T-cell activation and chemotaxis are decoupled using a HIV-1/SIVpbj1.9. chimeric nef variant. Arch. Virol. 158, 845-852 (2013).
Fortin, J. F., Barat, C., Beausejour, Y., Barbeau, B. & Tremblay, M. J. Hyper-responsiveness to stimulation of human immunodeficiency virus-infected CD4+ T cells requires Nef and Tat virus gene products and results from higher NFAT, NF-kappaB, and AP-1 induction. J. Biol. Chem. 279, 39520-39531 (2004).
Doyon, G., Zerbato, J., Mellors, J. W. & Sluis-Cremer, N. Disulfiram reactivates latent HIV-1 expression through depletion of the phosphatase and tensin homolog. AIDS 27, F7-F11 (2013).
Gupta, A. K. et al. The HIV protease inhibitor nelfinavir downregulates Akt phosphorylation by inhibiting proteasomal activity and inducing the unfolded protein response. Neoplasia 9, 271-278 (2007).
Cummins, N. W., Sainski, A. M., Natesampillai, S., Bren, G. D. & Badley, A. D. Choice of antiretroviral therapy differentially impacts survival of HIV-infected CD4 T cells. Mol. Cell Ther 2, 1 (2014).
Titanji, B. K., Aasa-Chapman, M., Pillay, D. & Jolly, C. Protease inhibitors effectively block cell-to-cell spread of HIV-1 between T cells. Retrovirology 10, 161 (2013).
Tyagi, M. & Bukrinsky, M. Human immunodeficiency virus (HIV) latency: the major hurdle in HIV eradication. Mol. Med. 18, 1096-1108 (2012).
Matsuda, Y., Kobayashi-Ishihara, M., Fujikawa, D., Ishida, T., Watanabe, T. & Yamagishi, M. Epigenetic heterogeneity in HIV-1 latency establishment. Sci Rep. 5, 7701 (2015).
Darcis, G. et al. An In-Depth Comparison of Latency-Reversing Agent Combinations in Various In Vitro and Ex Vivo HIV-1 Latency Models Identified Bryostatin-1+ JQ1 and Ingenol-B+ JQ1 to Potently Reactivate Viral Gene Expression. Plos. Pathog. 11, e1005063 (2015).
Kumar, A., Darcis, G., Van Lint, C. & Herbein, G. Epigenetic control of HIV-1 post integration latency: implications for therapy. Clinical Epigenetics 7, 103 (2015).
König, R. & Stertz, S. Recent strategies and progress in identifying host factors involved in virus replication. Curr Opin Microbiol. 26, 79-88 (2015).
Chugh, P. et al. Akt inhibitors as an HIV-1 infected macrophage-specific anti-viral therapy. Retrovirology 5, 11 (2008).
Herbein, G. et al. Apoptosis of CD8+ T cells is mediated by macrophages through interaction of HIV gp120 with chemokine receptor CXCR4. Nature 395, 189-194 (1998).
Popik, W., Alce, T. M. & Au, W. C. Human immunodeficiency virus type 1 uses lipid raft-colocalized CD4 and chemokine receptors for productive entry into CD4+ T cells. J Virol. 76, 4709-4722 (2002).
