Large-Scale Screening and Identification of Novel Ebola Virus and Marburg Virus Entry Inhibitors

Anantpadma, Manu; Kouznetsova, Jennifer; Wang, Hang; Huang, Ruili; Kolokoltsov, Andrey A.; Guha, Rajarshi; Lindstrom, Aaron R.; Shtanko, Olena; Simeonov, Anton; Maloney, David J.; Maury, Wendy; LaCount, Douglas; Jadhav, Ajit; Davey, Robert A.

doi:10.1128/aac.00543-16

Cited by 48 publications

(62 citation statements)

References 54 publications

(68 reference statements)

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“…EBOV ( Zaire ebolavirus, Mayinga variant) and recombinant EBOV expressing enhanced GFP (EBOV-EGFP) were grown and concentrated as described previously (Anantpadma et al, 2016). Briefly, virus was amplified in Vero cells in DMEM containing 2% FBS for 7 days.…”

Section: Star Methodsmentioning

confidence: 99%

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Protein Interaction Mapping Identifies RBBP6 as a Negative Regulator of Ebola Virus Replication

Batra

Hultquist

Liu

et al. 2018

Cell

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SUMMARY Ebola virus (EBOV) infection often results in fatal illness in humans, yet little is known about how EBOV usurps host pathways during infection. To address this, we used affinity tag-purification mass spectrometry (AP-MS) to generate an EBOV-host protein-protein interaction (PPI) map. We uncovered 194 high-confidence EBOV-human PPIs, including one between the viral transcription regulator VP30 and the host ubiquitin ligase RBBP6. Domain mapping identified a 23 amino acid region within RBBP6 that binds to VP30. A crystal structure of the VP30-RBBP6 peptide complex revealed that RBBP6 mimics the viral nucleoprotein (NP) binding to the same interface of VP30. Knockdown of endogenous RBBP6 stimulated viral transcription and increased EBOV replication, whereas overexpression of either RBBP6 or the peptide strongly inhibited both. These results demonstrate the therapeutic potential of biologies that target this interface and identify additional PPIs that may be leveraged for novel therapeutic strategies.

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Section: Star Methodsmentioning

confidence: 99%

“…24 h post-transfection, cells were carried to a BSL-4 lab and challenged with EBOV at an MOI of 0.5. Cells were fixed after 24 h in 10% neutral buffered formalin and stained for EBOV GP by using a previously described protocol (Anantpadma et al, 2016). Briefly, 24 h post-infection, cells were fixed by immersing the plate in formalin.…”

Section: Star Methodsmentioning

confidence: 99%

Protein Interaction Mapping Identifies RBBP6 as a Negative Regulator of Ebola Virus Replication

Batra

Hultquist

Liu

et al. 2018

Cell

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112

123

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“…mCherry-labeled Zaire viruslike particles (VLPs) were prepared and concentrated as described previously (36)(37)(38). Additionally, the conformation of Zaire GPs on the surface of VLPs was verified by an ELISA binding assay with MIL77 antibodies as mentioned above.…”

Section: Discussionmentioning

confidence: 99%

“…To determine which step may be the target for HP-HSA, fluorescent protein-labeled viruslike particles (VLPs) of ZEBOV (Zaire VLPs) were introduced. Recombinant EBOV VLPs, which are morphologically similar to authentic EBOV virions, have been extensively utilized in EBOV research, such as in studies of virion morphology and the viral entry mechanism and in drug development studies, as credible substitutes for infectious EBOVs (32,(36)(37)(38)(39)(40).…”

Section: Fig 3 Ex Vivomentioning

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Chemically Modified Human Serum Albumin Potently Blocks Entry of Ebola Pseudoviruses and Viruslike Particles

Xia

et al. 2017

Antimicrob Agents Chemother

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Ebola virus (EBOV), the causative pathogen of the deadly Ebola virus disease (EVD), can be transmitted via contact with EVD patients, including sexual contact with EVD survivors. At present, no licensed vaccine or therapeutic is available. In this study, we compared eight anhydride-modified proteins for their entry-inhibitory activity against the pseudovirus (PsV) carrying the envelope glycoprotein (GP) of the EBOV Zaire or Sudan species (Zaire PsV and Sudan PsV, respectively). We found that 3-hydroxyphthalic anhydride-modified human serum albumin (HP-HSA) was the most effective in inhibiting the entry of both Zaire PsV and Sudan PsV, with the 50% effective concentration being at the nanomolar level and with HP-HSA being more potent than EBOV-neutralizing antibody MIL77-2 (4G7, a component antibody of the ZMapp drug cocktail). The combination of HP-HSA and MIL77-2 exhibited a synergistic effect. HP-HSA had no obvious in vitro or in vivo toxicity. The EBOV PsV entryinhibitory activity of HP-HSA remained intact after storage at 45°C for 8 weeks, suggesting that HP-HSA has the potential for worldwide use, including tropical regions in African countries, as either a therapeutic to treat EBOV infection or a prophylactic microbicide to prevent the sexual transmission of EBOV.

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Phenotypic Prioritization of Diphyllin Derivatives That Block Filoviral Cell Entry by Vacuolar (H⁺)‐ATPase Inhibition

et al. 2018

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Many viruses utilize endosomal pathways to gain entry to cells and propagate infection. Sensing of endosomal acidification is a trigger for release of many virus cores into the cell cytosol. Previous efforts with inhibitors of vacuolar-ATPase have been shown to block endosomal acidification and affect viral entry albeit with limited potential for therapeutic selectivity. Herein, four novel series of derivatives of the vacuolar-ATPase inhibitor diphyllin were synthesized to assess their potential for enhancing potency and anti-filoviral activity over cytotoxicity. Derivatives that suitably blocked cellular entry of Ebola pseudo-typed virus were further evaluated as inhibitors of endosomal acidification and isolated human Vacuolar-ATPase activity. Several compounds with significant increases in potency over diphyllin in these assays also separated from cytotoxic doses in human cell models by >100 fold. Finally, three derivatives were shown to be inhibitors of replication-competent Ebola viral entry into primary macrophages with comparable potencies and enhanced selectivity toward anti-viral activity.

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Large-Scale Screening and Identification of Novel Ebola Virus and Marburg Virus Entry Inhibitors

Cited by 48 publications

References 54 publications

Protein Interaction Mapping Identifies RBBP6 as a Negative Regulator of Ebola Virus Replication

Protein Interaction Mapping Identifies RBBP6 as a Negative Regulator of Ebola Virus Replication

Chemically Modified Human Serum Albumin Potently Blocks Entry of Ebola Pseudoviruses and Viruslike Particles

Phenotypic Prioritization of Diphyllin Derivatives That Block Filoviral Cell Entry by Vacuolar (H⁺)‐ATPase Inhibition

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Large-Scale Screening and Identification of Novel Ebola Virus and Marburg Virus Entry Inhibitors

Cited by 48 publications

References 54 publications

Protein Interaction Mapping Identifies RBBP6 as a Negative Regulator of Ebola Virus Replication

Protein Interaction Mapping Identifies RBBP6 as a Negative Regulator of Ebola Virus Replication

Chemically Modified Human Serum Albumin Potently Blocks Entry of Ebola Pseudoviruses and Viruslike Particles

Phenotypic Prioritization of Diphyllin Derivatives That Block Filoviral Cell Entry by Vacuolar (H+)‐ATPase Inhibition

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Phenotypic Prioritization of Diphyllin Derivatives That Block Filoviral Cell Entry by Vacuolar (H⁺)‐ATPase Inhibition