Harmine treatment of PMA-treated J-Lat 5A8 cells
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE136172 | GEO | 2020/08/21
REPOSITORIES: GEO
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Project description:Harmine treatment of PMA-treated J-Lat 5A8 cells
| PRJNA561499 | ENA
Project description:HIV-1 latent reservoirs constitute a major barrier to viral eradication. The various potencies of latency-reversing agents reflect the multiplicity of the silencing mechanisms underlying latency, and the dynamic and heterogeneous nature of the HIV-1 latent reservoirs. Here, we investigated the mechanisms of the DNA methylation inhibitor 5-aza-2’-deoxycytidine (5-AzadC) potency in HIV-1 latency reversal. We report that this potency was directly linked to 5-AzadC-induced demethylation of the HIV-1 promoter, which occurred at non-random CpG dinucleotides. Specifically, the methylation status of one of those CpGs allowed the recruitment of the epigenetic integrator UHRF1 to the HIV-1 promoter. 5-AzadC-induced viral reactivation led to a decreased in vivo recruitment of UHRF1. RNA interference-mediated knockdown of UHRF1 demonstrated its role in DNA methylation-mediated silencing of the latent HIV-1 promoter. Pharmacological downregulation of UHRF1 in ex vivo HIV+ patient cell cultures resulted in potent reactivation of latent HIV-1, further highlighting that UHRF1 could constitute a new molecular target to devise innovative HIV-1 curative strategies.
2020-10-24 | GSE139320 | GEO
Project description:Despite effective treatment, HIV can persist in latent reservoirs, which represent a major obstacle towards HIV eradication. Targeting and reactivating latent cells is challenging due to the heterogeneous nature of HIV infected cells. Here, we used a primary model of HIV latency and single-cell RNA sequencing to characterize transcriptional heterogeneity during HIV latency and reactivation. Our analysis identified transcriptional programs leading to successful reactivation of HIV expression. We further validated our results using primary CD4+ T cells isolated from HIV+ individuals.
2018-03-13 | GSE111727 | GEO
Project description:HIV-1 eradication strategies require reactivation of all latent cells. Current methods lack effectiveness due to incomplete knowledge of proviral reactivation. We employed a single-molecule RNA-FISH (smRNA-FISH) analysis and found that proviral reactivation is highly variable, stochastic, and occurs in bursts and waves, with different kinetics in response to diverse LRAs. Approximately 1-5% of latent cells displayed stochastic reactivation even without stimulation. Through single-cell RNA-seq analysis, we identified NR4A3 and cMYC as extrinsic factors selectively associated with HIV-1 reactivation. Concomitant with HIV-1 reactivation cMYC was downregulated and NR4A3 was upregulated in both latent cell lines and primary CD4+ T-cells from aviremic patients. By inhibiting cMYC using SN-38, an active metabolite of irinotecan, we induced NR4A3 and HIV-1 expression. Our results suggest that inherent stochasticity in proviral reactivation contributes to cell-to-cell variability, which could potentially be modulated by drugs targeting cMYC and NR4A3.
2024-05-24 | GSE241207 | GEO
Project description:After stimulation of RAW264.7 cells with LPS and Harmine, high-throughput sequencing technology was used to explore the changes in the inflammation level of RAW cells and the enrichment of signaling pathways after LPS and Harmine treatment, and to find an interesting target pathway.
2024-05-20 | GSE267507 | GEO
Project description:The switch between HIV latency and productive transcription is regulated by an auto-feedback mechanism initiated by the viral trans-activator Tat, which functions to recruit the host transcription elongation factor P-TEFb to proviral HIV. A heterodimeric complex of CDK9 and one of three cyclin T subunits, P-TEFb is expressed at vanishingly low levels in latently infected memory CD4+ T cells and cellular mechanisms controlling its availability for proviral transcription are not well-defined. Using a well-characterized primary T-cell model of HIV latency alongside healthy donor memory CD4+ T cells, we sought to identify specific T-cell receptor (TCR) signaling pathways that regulate the generation of transcriptionally active P-TEFb, defined as the coordinate expression of cyclin T1 and phospho-Ser175 CDK9. Unexpectedly, cellular treatment with diacylglycerol-mimicking protein kinase C (PKC) agonists in the absence of intracellular calcium mobilization with an ionophore was sufficient to stimulate active P-TEFb expression and reactivate latent HIV with minimal cytotoxicity. Furthermore, inhibition-based experiments demonstrated that PKC agonists and TCR-mobilized diacylglycerol signal through MAP kinases ERK1/2 rather than through PKC to effect the reactivation of both P-TEFb and latent HIV. Single-cell RNA-seq analysis revealed that of the four known isoforms of RasGRP, the diacylglycerol-dependent Ras guanine nucleotide exchange factor RasGRP1 is by far the predominantly expressed isoform in memory CD4+ T cells and should therefore mediate the activation of ERK1/2 upon TCR co-stimulation or PKC agonist challenge. Combined inhibition of the PI3K-AKT-mTORC1/2 pathway alongside the ERK1/2 activator MEK prior to TCR co-stimulation abrogated active P-TEFb expression and strongly suppressed latent HIV reactivation. Therefore, contrary to prevailing models, the coordinate reactivation of P-TEFb and latent HIV in primary T cells in response to either TCR co-stimulation or PKC agonist challenge is largely independent of PKC activity but rather primarily involves two complementary signaling arms of the TCR cascade namely RasGRP1-Ras-Raf-MEK-ERK1/2 and PI3K-AKT-mTORC1/2.
2021-09-07 | GSE167916 | GEO
Project description:The barrier to HIV-1 functional cure is caused by a small pool of latently infected resting CD4 T-cells that persist under antiretroviral therapy. Notably this latent reservoir of infected cells will produce replication-competent infectious virus once prolonged suppressive HAART is withdrawn. The reactivation of HIV-1 gene expression in T-cells harboring latent provirus in HIV-1 patients under HAART will likely result in depletion of this latent reservoir due to cytopathic effects and immune clearance. Many studies have investigated small molecules that reactivate HIV-1 gene expression but to date no latency reversal agent (LRA) has been identified to be specific, non-toxic, and effective in primary T-cells isolated from HIV-1 infected individuals undergoing long-term HAART. Stochastic fluctuations in HIV-1 tat gene expression have been attributed to be essential in the viral progression to latency. We hypothesized that exposing Tat to latently infected CD4 T-cells will result in potent latency reversal. Our results indicate the capacity of an engineered Tat to reactivate HIV-1 in latently infected cells from patients to a similar degree as the protein kinase C agonist PMA (Phorbol 12-Myristate 13-Acetate) while showing no T-cell activation nor any significant transcriptome perturbation in primary CD4 T-cells.
2023-09-05 | GSE235229 | GEO
Project description:RNA-seq was applied for identifying the change of transcriptome between U2OS cells with starvation-induced quiescence treated with Harmine Hydrochloride or INDY and the untreated controls.
2023-02-01 | GSE168401 | GEO
Project description:A major obstacle for HIV eradication is the persistence of latent reservoirs, cells harboring replication competent yet transcriptionally silent proviruses. These reservoirs, composed primarily of CD4+ T cells, cannot be targeted by the host immune system or by combination anti-retroviral therapy (cART). Upon cessation of cART, a rapid viral rebound occurs. In order to develop effective HIV cure therapies, a better understanding of the factors and host programd governing latency establishment and/or maintenance must be obtained. To achieve this goal, we treated a CD4+T cell-based model of latency (J-Lat 10.6) with Latency Reversing Agents (LRAs) and performed bulk RNA-seq to predict host transcriptional programs activated, and potentially involved, in latent HIV reactivation. Our findings suggest that a novel small molecule (EPH-334) reactivates latent HIV by activating an oxidative stress transcriptional program linked to activation of MAPKs and downstream master regulators. Interestingly, the pan histone deacetylase inhibitor SAHA also induces oxidative stress programs potentially linking sensing of redox state aletartions with activation of cell signaling and transcriptioonal programs culminating on latent HIV reactivation.
2020-07-30 | GSE144552 | GEO
Project description:HIV-1 latency is a critical hurdle to a cure for HIV-1 infection, but our understanding of the molecular biology of latency control is still incomplete. We provide experimental evidence that HIV-1 infection of primary T cells and T cell lines generates a substantial amount of TCR/CD3 activation-inert latently infected T cells. Proteomic and genome-wide RNA-level analysis comparing CD3-responsive and CD3-inert latently HIV-1 infected T cells, followed by software-based data integration suggested two phenomena to govern CD3-inertness: (i) the presence of extensive transcriptomic noise that affected the efficacy of CD3 signaling and (ii) defined changes to specific signaling pathways. Validation experiments demonstrated that compounds known to increase transcriptomic noise further diminished the ability of TCR/CD3 stimulation to trigger HIV-1 reactivation. Conversely, targeting specific central nodes in the generated PINs such as STAT3 improved the ability of TCR/CD3 activation to trigger HIV-1 reactivation in T cell lines and primary T cells. The data emphasize that latent HIV-1 infection is largely the result of extensive, stable biomolecular changes to the signaling network of the host T cells harboring latent HIV-1 infection events. We discuss the implications of these findings for the idea of using single drug therapies to trigger HIV-1 reactivation in the latent T cell reservoir in patients.
2020-06-19 | GSE152788 | GEO