Gliotoxin, identified from a screen of fungal metabolites, disrupts 7SK snRNP, releases P-TEFb, and reverses HIV-1 latency

Stoszko, Mateusz; Al‐Hatmi, Abdullah M. S.; Škríba, Anton; Röling, Michael D.; Ne, Enrico; Crespo, Raquel; Mueller, Yvonne M.; Najafzadeh, Mohammad Javad; Kang, Je-Won; Ptackova, Renata; LeMasters, E.; Biswas, Pritha; Bertoldi, Alessia; Kan, Tsung Wai; Crignis, Elisa De; Šulc, Miroslav; Lebbink, Joyce H.G.; Rokx, Casper; Verbon, Annelies; IJcken, Wilfred F. J. van; Katsikis, Peter D.; Palstra, Robert-Jan; Havlı́ček, Vladimı́r; Hoog, Sybren de; Mahmoudi, Tokameh

doi:10.1126/sciadv.aba6617

Cited by 14 publications

(21 citation statements)

References 65 publications

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“…Topiramate treatment did not significantly affect CD4 + T cell viability after 48 h, while control treatment with a toxic (200 nM) concentration of gliotoxin (GTX) caused apoptosis of primary CD4 + T cells, as evidenced by an increase in the percentage of annexin V-positive cells, as was observed previously ( Fig. 4E ) ( 78 ). In contrast to what was observed in the A2 and 11.1 J-Lat cell lines, treatment of latently infected CD4 + T cells with the NF1 inhibitor trametinib did not lead to a significant reversal of latency ( Fig.…”

Section: Resultssupporting

confidence: 76%

A Two-Color Haploid Genetic Screen Identifies Novel Host Factors Involved in HIV-1 Latency

Röling

Sisakht

et al. 2021

mBio

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Section: Resultssupporting

confidence: 76%

A Two-Color Haploid Genetic Screen Identifies Novel Host Factors Involved in HIV-1 Latency

Röling

Sisakht

et al. 2021

mBio

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“…Treatment with the GRIK5 inhibitor Topiramate, at a concentration of 16 and 20 µM, resulted in significant (p < 0.05) reversal of latency, as measured by a 3-6 fold increases in mean luciferase activity compared to untreated controls (Figure 4D). Topiramate treatment did not significantly affect CD4+ T cell viability after 48 hours, while control treatment with a toxic (200nM) concentration of Gliotoxin (GTX) caused apoptosis of primary CD4+ T-cells, as evidenced by an increase in the percentage of Annexin V-positive cells, as was observed previously (Figure 4E) (80). In contrast to what was observed in the A2 and 11.1 J-Lat cell-lines, treatment of latently infected CD4+ T-cells with the NF1 inhibitor, Trametinib did not lead to a significant reversal of latency (Figure 4D), while viability was only moderately affected (figure 4E).…”

Section: Resultssupporting

confidence: 74%

A two-color haploid genetic screen identifies novel host factors involved in HIV latency

Röling

Sisakht

et al. 2021

Preprint

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To identify novel host factors as putative targets to reverse HIV latency, we performed an insertional mutagenesis genetic screen in a latently HIV-1-infected pseudo-haploid KBM7 cell line (Hap-Lat). Following mutagenesis, insertions were mapped to the genome and bioinformatic analysis resulted in the identification of 69 candidate host genes involved in maintaining HIV-1 latency. A select set of candidate genes was functionally validated using shRNA mediated depletion in latent HIV-1 infected J-Lat A2 and 11.1 T cell lines. We confirmed ADK, CHD9, CMSS1, EVI2B, EXOSC8, FAM19A, GRIK5, IRF2BP2, NF1, and USP15 as novel host factors involved in the maintenance of HIV latency. Chromatin immunoprecipitation assays indicated that CHD9, a Chromodomain Helicase DNA-binding protein, maintains HIV latency via direct association with the HIV 5’LTR, and its depletion results in increased histone acetylation at the HIV-1 promoter, concomitant with HIV-1 latency reversal. FDA-approved inhibitors 5-Iodotubercidin, Trametinib, and Topiramate, targeting ADK, NF1, and GRIK5, respectively were characterized for their latency reversal potential. While 5-Iodotubercidin exhibited significant cytotoxicity in both J-Lat and primary CD4+ T cells, Trametinib reversed latency in J-Lat cells but not in latently HIV-1-infected primary CD4+ T cells. Crucially, Topiramate reversed latency in cell-line models and latently infected primary CD4+ T cells, without inducing T cell activation or significant toxicity. Thus, using an adaptation of a haploid forward genetic screen, we identified novel and druggable host factors contributing to HIV-1 latency.ImportanceA reservoir of latent HIV-1-infected cells persists in the presence of combination antiretroviral therapy (cART), representing a major obstacle for viral eradication. Reactivation of the latent HIV-1 provirus is part of curative strategies which aim to promote clearance of the infected cells. Using a two-color haploid screen, we identified 69 candidate genes as latency maintaining host factors and functionally validated a subset of 10 of those in additional T-cell based cell line models of HIV-1 latency. We further demonstrated that CHD9 is associated with HIV-1’s promoter, the 5’LTR while this association is lost upon reactivation. Additionally, we characterized the latency reversal potential of FDA compounds targeting ADK, NF1, and GRIK5 and identify the GRIK5 inhibitor Topiramate as a viable latency reversal agent with clinical potential.

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“…In our investigation, the interactions outside of the active site are also reported by using the blind docking methodology available online at Blind Docking (BD) Server. This software was used to improve the knowledge about molecular pathways of auto inflammatory diseases [15], also this technique was utilized in infectious diseases including the discovery of new possible treatments against virus like HIV-1 or SARS-CoV-2 [16][17]. More interaction points reported by the present study may contribute further support to the idea of development of C.cylindracea's secondary metabolite-based SARS-CoV-2 neutralizing agents predicted by BD technique [18].…”

Section: Introductionmentioning

confidence: 52%

Secondary Metabolites from Caulerpa Cylindracea (Sonder) Could Be Alternative Natural Antiviral Compounds for COVID-19: A Further in Silico Proof

Çavaş¹,

C²,

Carmena-Bargueño³

et al. 2020

Preprint

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<p>SARS-CoV-2 has been exhibiting extremely spreading property all around the world since its existence from Wuhan-China in December-2019. Although it has caused a death toll of over than 1.3 M people, no validated vaccine has been proposed yet. On the other hand, very dense studies on the vaccine development have been carrying out in some countries such as the US, Germany, UK, China and Russia. Due to side effects of current antiviral agents used in the therapy of COVID-19, there is a great need for the development of alternative compounds for this disease. Caulerpin (CPN) and caulerpenyne (CYN), predominant natural secondary metabolites from invasive marine green algae <i>Caulerpa cylindracea,</i>are proposed to neutralize the virus from two targets: spike protein (5XLR) and main protease (6YB7) in this study. The results show that the binding energies related to CPN-6YB7 and CYN-6YB7 interactions are found to be -8.02 kcal/mol and -6.83 kcal/mol, respectively. The binding energies were -9.68 kcal/mol and -7.53 kcal/mol, respectively, for CPN-5XLR and CYN-5XLR. In the molecular dynamics results, RMSD values show that CPN and CYN can form stable complexes with the proteins where CYN is more stable with 6YB7 and CPN interacts better with 5XLR. These differences seem to be based on the type of interactions of the complexes. In conclusion, caulerpin and caulerpenyne can further be investigated experimentally for their anti-SARS-CoV-2 efficiency. </p>

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Gliotoxin, identified from a screen of fungal metabolites, disrupts 7SK snRNP, releases P-TEFb, and reverses HIV-1 latency

Cited by 14 publications

References 65 publications

A Two-Color Haploid Genetic Screen Identifies Novel Host Factors Involved in HIV-1 Latency

A Two-Color Haploid Genetic Screen Identifies Novel Host Factors Involved in HIV-1 Latency

A two-color haploid genetic screen identifies novel host factors involved in HIV latency

Secondary Metabolites from Caulerpa Cylindracea (Sonder) Could Be Alternative Natural Antiviral Compounds for COVID-19: A Further in Silico Proof

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