2. Academic Validation
  3. High-throughput split-GFP antiviral screening assay against fusogenic paramyxoviruses

High-throughput split-GFP antiviral screening assay against fusogenic paramyxoviruses

  • Antiviral Res. 2025 Sep:241:106242. doi: 10.1016/j.antiviral.2025.106242.
Laura Vandemaele 1,  Thibault Francken 1,  Joost Schepers 1,  Winston Chiu 1,  Niels Cremers 1,  Hugo Klaassen 2,  Charlène Marcadet 2,  Lorena Sanchez Felipe 3,  Arnaud Marchand 2,  Patrick Chaltin 4,  Pieter Leyssen 1,  Johan Neyts 5,  Manon Laporte 1
Affiliations

Affiliations

  • 1 Department of Microbiology, Immunology and Transplantation, Rega Institute, Antiviral Drug & Vaccine Research Group, KU Leuven, Herestraat 49 - box 1043, 3000, Leuven, Belgium.
  • 2 Cistim Leuven vzw, Bioincubator 2, Gaston Geenslaan 2, 3001, Leuven, Belgium.
  • 3 Department of Microbiology, Immunology and Transplantation, VirusBank Platform, KU Leuven, Arenberg Accelerator, Gaston Geenslaan 3, 3001, Leuven, Belgium.
  • 4 Cistim Leuven vzw, Bioincubator 2, Gaston Geenslaan 2, 3001, Leuven, Belgium; Center for Drug Design and Development (CD3), KU Leuven R&D, Waaistraat 6, B-3000, Leuven, Belgium.
  • 5 Department of Microbiology, Immunology and Transplantation, Rega Institute, Antiviral Drug & Vaccine Research Group, KU Leuven, Herestraat 49 - box 1043, 3000, Leuven, Belgium; Department of Microbiology, Immunology and Transplantation, VirusBank Platform, KU Leuven, Arenberg Accelerator, Gaston Geenslaan 3, 3001, Leuven, Belgium. Electronic address: johan.neyts@kuleuven.be.
PMID: 40701276 DOI: 10.1016/j.antiviral.2025.106242
Abstract

The paramyxovirus family includes important pathogens such as measles and mumps viruses, as well as emerging pathogens with pandemic potential such as Nipah virus. Despite the threat to public health and the frequent identification of novel paramyxoviruses, no Antiviral drugs are currently available. A hallmark of most paramyxoviruses is the induction of cell-cell fusion leading to syncytia formation. To facilitate Antiviral drug discovery, we leveraged this trait and established a high-throughput split-green fluorescent protein (GFP) Antiviral screening assay suitable for high-content imaging through the quantification of virus-induced GFP+ syncytia. The assay was validated with well-known broad-spectrum Antiviral compounds against representative members of five different Paramyxovirinae genera. Using this split-GFP assay, a small-molecule repurposing library of approximately 3000 compounds was screened against recombinant Cedar virus (CedV), a nonpathogenic henipavirus. Two molecules were identified: Cathepsin Inhibitor 1 with henipavirus-specific activity and PF-543 with pan-paramyxovirus activity. Both molecules inhibit viral replication by blocking cell-cell fusion. The split-GFP assay presented here will enable the development of extensive drug discovery initiatives aimed at identifying much-needed pan-henipavirus/paramyxovirus inhibitors.

Figures
Products