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A High-Throughput HIV-1 Drug Screening Platform, Based on Lentiviral Vectors and Compatible with Biosafety Level-1

HIV-1 infection is a complex, multi-step process involving not only viral, but also multiple cellular factors. To date, drug discovery methods have primarily focused on the inhibition of single viral proteins. We present an efficient and unbiased approach, compatible with biosafety level 1 (BSL-1) c...

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Autores principales: Ellinger, Bernhard, Pohlmann, Daniel, Woens, Jannis, Jäkel, Felix M., Reinshagen, Jeanette, Stocking, Carol, Prassolov, Vladimir S., Fehse, Boris, Riecken, Kristoffer
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7290285/
https://www.ncbi.nlm.nih.gov/pubmed/32466195
http://dx.doi.org/10.3390/v12050580
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author Ellinger, Bernhard
Pohlmann, Daniel
Woens, Jannis
Jäkel, Felix M.
Reinshagen, Jeanette
Stocking, Carol
Prassolov, Vladimir S.
Fehse, Boris
Riecken, Kristoffer
author_facet Ellinger, Bernhard
Pohlmann, Daniel
Woens, Jannis
Jäkel, Felix M.
Reinshagen, Jeanette
Stocking, Carol
Prassolov, Vladimir S.
Fehse, Boris
Riecken, Kristoffer
author_sort Ellinger, Bernhard
collection PubMed
description HIV-1 infection is a complex, multi-step process involving not only viral, but also multiple cellular factors. To date, drug discovery methods have primarily focused on the inhibition of single viral proteins. We present an efficient and unbiased approach, compatible with biosafety level 1 (BSL-1) conditions, to identify inhibitors of HIV-1 reverse transcription, intracellular trafficking, nuclear entry and genome integration. Starting with a fluorescent assay setup, we systematically improved the screening methodology in terms of stability, efficiency and pharmacological relevance. Stability and throughput were optimized by switching to a luciferase-based readout. BSL-1 compliance was achieved without sacrificing pharmacological relevance by using lentiviral particles pseudo-typed with the mouse ecotropic envelope protein to transduce human PM1 T cells gene-modified to express the corresponding murine receptor. The cellular assay was used to screen 26,048 compounds selected for maximum diversity from a 200,640-compound in-house library. This yielded z’ values greater than 0.8 with a hit rate of 3.3% and a confirmation rate of 50%. We selected 93 hits and enriched the collection with 279 similar compounds from the in-house library to identify promising structural features. The most active compounds were validated using orthogonal assay formats. The similarity of the compound profiles across the different platforms demonstrated that the reported lentiviral assay system is a robust and versatile tool for the identification of novel HIV-1 inhibitors.
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spelling pubmed-72902852020-06-15 A High-Throughput HIV-1 Drug Screening Platform, Based on Lentiviral Vectors and Compatible with Biosafety Level-1 Ellinger, Bernhard Pohlmann, Daniel Woens, Jannis Jäkel, Felix M. Reinshagen, Jeanette Stocking, Carol Prassolov, Vladimir S. Fehse, Boris Riecken, Kristoffer Viruses Article HIV-1 infection is a complex, multi-step process involving not only viral, but also multiple cellular factors. To date, drug discovery methods have primarily focused on the inhibition of single viral proteins. We present an efficient and unbiased approach, compatible with biosafety level 1 (BSL-1) conditions, to identify inhibitors of HIV-1 reverse transcription, intracellular trafficking, nuclear entry and genome integration. Starting with a fluorescent assay setup, we systematically improved the screening methodology in terms of stability, efficiency and pharmacological relevance. Stability and throughput were optimized by switching to a luciferase-based readout. BSL-1 compliance was achieved without sacrificing pharmacological relevance by using lentiviral particles pseudo-typed with the mouse ecotropic envelope protein to transduce human PM1 T cells gene-modified to express the corresponding murine receptor. The cellular assay was used to screen 26,048 compounds selected for maximum diversity from a 200,640-compound in-house library. This yielded z’ values greater than 0.8 with a hit rate of 3.3% and a confirmation rate of 50%. We selected 93 hits and enriched the collection with 279 similar compounds from the in-house library to identify promising structural features. The most active compounds were validated using orthogonal assay formats. The similarity of the compound profiles across the different platforms demonstrated that the reported lentiviral assay system is a robust and versatile tool for the identification of novel HIV-1 inhibitors. MDPI 2020-05-25 /pmc/articles/PMC7290285/ /pubmed/32466195 http://dx.doi.org/10.3390/v12050580 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ellinger, Bernhard
Pohlmann, Daniel
Woens, Jannis
Jäkel, Felix M.
Reinshagen, Jeanette
Stocking, Carol
Prassolov, Vladimir S.
Fehse, Boris
Riecken, Kristoffer
A High-Throughput HIV-1 Drug Screening Platform, Based on Lentiviral Vectors and Compatible with Biosafety Level-1
title A High-Throughput HIV-1 Drug Screening Platform, Based on Lentiviral Vectors and Compatible with Biosafety Level-1
title_full A High-Throughput HIV-1 Drug Screening Platform, Based on Lentiviral Vectors and Compatible with Biosafety Level-1
title_fullStr A High-Throughput HIV-1 Drug Screening Platform, Based on Lentiviral Vectors and Compatible with Biosafety Level-1
title_full_unstemmed A High-Throughput HIV-1 Drug Screening Platform, Based on Lentiviral Vectors and Compatible with Biosafety Level-1
title_short A High-Throughput HIV-1 Drug Screening Platform, Based on Lentiviral Vectors and Compatible with Biosafety Level-1
title_sort high-throughput hiv-1 drug screening platform, based on lentiviral vectors and compatible with biosafety level-1
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7290285/
https://www.ncbi.nlm.nih.gov/pubmed/32466195
http://dx.doi.org/10.3390/v12050580
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