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Open AccessArticle

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

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Department ScreeningPort, Fraunhofer Institute for Molecular Biology and Applied Ecology IME, 22525 Hamburg, Germany
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Fraunhofer Cluster of Excellence Immune-Mediated Diseases CIMD, Partner site Hamburg, 22525 Hamburg, Germany
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Research Department Cell and Gene Therapy, Department of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf (UKE), 20246 Hamburg, Germany
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Heinrich-Pette-Institute, Leibniz Institute for Experimental Virology, 20251 Hamburg, Germany
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Engelhardt-Institute of Molecular Biology, Russian Academy of Sciences, 117984 Moscow, Russia
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German Center for Infection Research (DZIF), Partner site Hamburg, 20246 Hamburg, Germany
*
Authors to whom correspondence should be addressed.
Viruses 2020, 12(5), 580; https://doi.org/10.3390/v12050580
Received: 29 April 2020 / Revised: 19 May 2020 / Accepted: 20 May 2020 / Published: 25 May 2020
(This article belongs to the Special Issue Antiretroviral Drug Development and HIV Cure Research)
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. View Full-Text
Keywords: HIV-1 drug development; BSL-1 screening platform; high-throughput screening; lentiviral vectors; mCat1 expressing PM1 T cell line; LeGO vectors HIV-1 drug development; BSL-1 screening platform; high-throughput screening; lentiviral vectors; mCat1 expressing PM1 T cell line; LeGO vectors
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MDPI and ACS Style

Ellinger, B.; Pohlmann, D.; Woens, J.; Jäkel, F.M.; Reinshagen, J.; Stocking, C.; Prassolov, V.S.; Fehse, B.; Riecken, K. A High-Throughput HIV-1 Drug Screening Platform, Based on Lentiviral Vectors and Compatible with Biosafety Level-1. Viruses 2020, 12, 580.

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