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Synergistic Reactivation of Latent HIV Expression by Ingenol-3-Angelate, PEP005, Targeted NF-kB Signaling in Combination with JQ1 Induced p-TEFb Activation.

Jiang G, Mendes EA, Kaiser P, Wong DP, Tang Y, Cai I, Fenton A, Melcher GP, Hildreth JE, Thompson GR, Wong JK, Dandekar S - PLoS Pathog. (2015)

Bottom Line: Therefore, effective strategies for eradication of the HIV reservoirs are urgently needed.Furthermore, PEP005 and the P-TEFb agonist, JQ1, exhibited synergism in reactivation of latent HIV with a combined effect that is 7.5-fold higher than the effect of PEP005 alone.This anti-cancer compound is a potential candidate for advancing HIV eradication strategies.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology and Immunology, University of California, Davis, Davis, California, United States of America.

ABSTRACT
Although anti-retroviral therapy (ART) is highly effective in suppressing HIV replication, it fails to eradicate the virus from HIV-infected individuals. Stable latent HIV reservoirs are rapidly established early after HIV infection. Therefore, effective strategies for eradication of the HIV reservoirs are urgently needed. We report that ingenol-3-angelate (PEP005), the only active component in a previously FDA approved drug (PICATO) for the topical treatment of precancerous actinic keratosis, can effectively reactivate latent HIV in vitro and ex vivo with relatively low cellular toxicity. Biochemical analysis showed that PEP005 reactivated latent HIV through the induction of the pS643/S676-PKCδ/θ-IκBα/ε-NF-κB signaling pathway. Importantly, PEP005 alone was sufficient to induce expression of fully elongated and processed HIV RNAs in primary CD4+ T cells from HIV infected individuals receiving suppressive ART. Furthermore, PEP005 and the P-TEFb agonist, JQ1, exhibited synergism in reactivation of latent HIV with a combined effect that is 7.5-fold higher than the effect of PEP005 alone. Conversely, PEP005 suppressed HIV infection of primary CD4+ T cells through down-modulation of cell surface expression of HIV co-receptors. This anti-cancer compound is a potential candidate for advancing HIV eradication strategies.

No MeSH data available.


Related in: MedlinePlus

PEP005 causes minimal cytotoxicity and T cell proliferation.To evaluate the impact of PEP005 on the cell viability, U1 cells (A), J-Lat A1 cells (B) and CD4+ T cells (C) were treated with 12 nM of PEP005 for 24 hours, the cell viability was examined with the MTT assay and the S-phase cell cycle progression was evaluated after BrdU incorporation using BrdU ELISA. 25–50 μM Etoposide or 100–150 μM Doxorubicin (Doxo) served as positive controls for MTT and BrdU assays respectively [48–50]. Statistical analysis was performed in comparison with controls. **, p<0.01.
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ppat.1005066.g007: PEP005 causes minimal cytotoxicity and T cell proliferation.To evaluate the impact of PEP005 on the cell viability, U1 cells (A), J-Lat A1 cells (B) and CD4+ T cells (C) were treated with 12 nM of PEP005 for 24 hours, the cell viability was examined with the MTT assay and the S-phase cell cycle progression was evaluated after BrdU incorporation using BrdU ELISA. 25–50 μM Etoposide or 100–150 μM Doxorubicin (Doxo) served as positive controls for MTT and BrdU assays respectively [48–50]. Statistical analysis was performed in comparison with controls. **, p<0.01.

Mentions: In order to further evaluate the effects of PEP005 on T cell activation, we determined the ability of PEP005 to induce T cell proliferation and cytotoxicity using BrdU incorporation and MTT assays. PEP005 induced minimal levels of cellular toxicity in both J-Lat A1 and U1 cell lines, as well as in primary CD4+ T cells from peripheral blood samples of healthy HIV-negative donors (Fig 7). Importantly, PEP005 treatment did not induce any significant increase in the proportion of cells in S-phase with J-Lat A1 cells, U1 cells or primary CD4+ T cells. In summary, despite the increased CD69 expression, minimal to no induction was found for the expression of pro-inflammatory cytokines and no significant impact was seen on cell cycling, suggesting that PEP005 may be a potential LRA candidate for evaluation in vivo.


Synergistic Reactivation of Latent HIV Expression by Ingenol-3-Angelate, PEP005, Targeted NF-kB Signaling in Combination with JQ1 Induced p-TEFb Activation.

Jiang G, Mendes EA, Kaiser P, Wong DP, Tang Y, Cai I, Fenton A, Melcher GP, Hildreth JE, Thompson GR, Wong JK, Dandekar S - PLoS Pathog. (2015)

PEP005 causes minimal cytotoxicity and T cell proliferation.To evaluate the impact of PEP005 on the cell viability, U1 cells (A), J-Lat A1 cells (B) and CD4+ T cells (C) were treated with 12 nM of PEP005 for 24 hours, the cell viability was examined with the MTT assay and the S-phase cell cycle progression was evaluated after BrdU incorporation using BrdU ELISA. 25–50 μM Etoposide or 100–150 μM Doxorubicin (Doxo) served as positive controls for MTT and BrdU assays respectively [48–50]. Statistical analysis was performed in comparison with controls. **, p<0.01.
© Copyright Policy
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4520526&req=5

ppat.1005066.g007: PEP005 causes minimal cytotoxicity and T cell proliferation.To evaluate the impact of PEP005 on the cell viability, U1 cells (A), J-Lat A1 cells (B) and CD4+ T cells (C) were treated with 12 nM of PEP005 for 24 hours, the cell viability was examined with the MTT assay and the S-phase cell cycle progression was evaluated after BrdU incorporation using BrdU ELISA. 25–50 μM Etoposide or 100–150 μM Doxorubicin (Doxo) served as positive controls for MTT and BrdU assays respectively [48–50]. Statistical analysis was performed in comparison with controls. **, p<0.01.
Mentions: In order to further evaluate the effects of PEP005 on T cell activation, we determined the ability of PEP005 to induce T cell proliferation and cytotoxicity using BrdU incorporation and MTT assays. PEP005 induced minimal levels of cellular toxicity in both J-Lat A1 and U1 cell lines, as well as in primary CD4+ T cells from peripheral blood samples of healthy HIV-negative donors (Fig 7). Importantly, PEP005 treatment did not induce any significant increase in the proportion of cells in S-phase with J-Lat A1 cells, U1 cells or primary CD4+ T cells. In summary, despite the increased CD69 expression, minimal to no induction was found for the expression of pro-inflammatory cytokines and no significant impact was seen on cell cycling, suggesting that PEP005 may be a potential LRA candidate for evaluation in vivo.

Bottom Line: Therefore, effective strategies for eradication of the HIV reservoirs are urgently needed.Furthermore, PEP005 and the P-TEFb agonist, JQ1, exhibited synergism in reactivation of latent HIV with a combined effect that is 7.5-fold higher than the effect of PEP005 alone.This anti-cancer compound is a potential candidate for advancing HIV eradication strategies.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Microbiology and Immunology, University of California, Davis, Davis, California, United States of America.

ABSTRACT
Although anti-retroviral therapy (ART) is highly effective in suppressing HIV replication, it fails to eradicate the virus from HIV-infected individuals. Stable latent HIV reservoirs are rapidly established early after HIV infection. Therefore, effective strategies for eradication of the HIV reservoirs are urgently needed. We report that ingenol-3-angelate (PEP005), the only active component in a previously FDA approved drug (PICATO) for the topical treatment of precancerous actinic keratosis, can effectively reactivate latent HIV in vitro and ex vivo with relatively low cellular toxicity. Biochemical analysis showed that PEP005 reactivated latent HIV through the induction of the pS643/S676-PKCδ/θ-IκBα/ε-NF-κB signaling pathway. Importantly, PEP005 alone was sufficient to induce expression of fully elongated and processed HIV RNAs in primary CD4+ T cells from HIV infected individuals receiving suppressive ART. Furthermore, PEP005 and the P-TEFb agonist, JQ1, exhibited synergism in reactivation of latent HIV with a combined effect that is 7.5-fold higher than the effect of PEP005 alone. Conversely, PEP005 suppressed HIV infection of primary CD4+ T cells through down-modulation of cell surface expression of HIV co-receptors. This anti-cancer compound is a potential candidate for advancing HIV eradication strategies.

No MeSH data available.


Related in: MedlinePlus