A three-dimensional high throughput assay identifies novel antibacterial molecules with activity against intracellular Shigella
Abstract The Gram-negative bacterial species Shigella is the second leading cause of diarrhea among children in low and middle-income countries (LMICs) and is a World Health Organization (WHO) priority pathogen. Shigella infections are becoming increasing difficult to treat due to antimicrobial resi...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-05-01
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| Series: | npj Antimicrobials and Resistance |
| Online Access: | https://doi.org/10.1038/s44259-025-00110-6 |
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| author | Voong Vinh Phat Andrew Shih Teong Lim Cristina De Cozar-Gallardo Maria Isabel Castellote Alvaro Demetrio Muñoz Alvarez Elena Fernandez Alvaro Lluis Ballell-Pages Sonia Lozano-Arias Stephen Baker |
| author_facet | Voong Vinh Phat Andrew Shih Teong Lim Cristina De Cozar-Gallardo Maria Isabel Castellote Alvaro Demetrio Muñoz Alvarez Elena Fernandez Alvaro Lluis Ballell-Pages Sonia Lozano-Arias Stephen Baker |
| author_sort | Voong Vinh Phat |
| collection | DOAJ |
| description | Abstract The Gram-negative bacterial species Shigella is the second leading cause of diarrhea among children in low and middle-income countries (LMICs) and is a World Health Organization (WHO) priority pathogen. Shigella infections are becoming increasing difficult to treat due to antimicrobial resistance (AMR), leading to an urgent need for new antimicrobial agents with novel modes of action. Shigella pathogenesis is largely intracellular and antibacterial chemicals that preferentially work inside cells may be desirable to limit collateral AMR and block key components of the Shigella infection cycle. Aiming to facilitate the process of identifying antibacterial chemicals that kill intracellular Shigella, we developed a high-throughput screening (HTS) cell-based chemical screening assay. The three-dimensional (3-D) assay, incorporating Shigella invasion into Caco-2 cells on Cytodex 3 beads, was scaled into a 384-well platform for screening chemical compound libraries. Using this assay, we evaluated >500,000 compounds, identifying 12 chemical hits that inhibit Shigella replication inside cells. This simple, efficient and HTS-compatible assays circumvents many of the limitations of traditional screening methods with cell monolayers and may be deployed for antibacterial compound screening for other intracellular pathogens. |
| format | Article |
| id | doaj-art-e0b163949084421c93eaf683e5b69964 |
| institution | OA Journals |
| issn | 2731-8745 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | npj Antimicrobials and Resistance |
| spelling | doaj-art-e0b163949084421c93eaf683e5b699642025-08-20T01:51:36ZengNature Portfolionpj Antimicrobials and Resistance2731-87452025-05-01311810.1038/s44259-025-00110-6A three-dimensional high throughput assay identifies novel antibacterial molecules with activity against intracellular ShigellaVoong Vinh Phat0Andrew Shih Teong Lim1Cristina De Cozar-Gallardo2Maria Isabel Castellote Alvaro3Demetrio Muñoz Alvarez4Elena Fernandez Alvaro5Lluis Ballell-Pages6Sonia Lozano-Arias7Stephen Baker8The Hospital for Tropical Diseases, Wellcome Trust Major Overseas Programme, Oxford University Clinical Research UnitThe ALBORADA Drug Discovery Institute, University of CambridgeGSK Global Health, Tres CantosMolecular Modalities Discovery, GSK, Tres CantosGSK Global Health, Tres CantosGSK Global Health, Tres CantosGSK Global Health, Tres CantosGSK Global Health, Tres CantosA*STAR Infectious Diseases Labs (A*STAR IDL), Agency for Science, Technology and Research (A*STAR)Abstract The Gram-negative bacterial species Shigella is the second leading cause of diarrhea among children in low and middle-income countries (LMICs) and is a World Health Organization (WHO) priority pathogen. Shigella infections are becoming increasing difficult to treat due to antimicrobial resistance (AMR), leading to an urgent need for new antimicrobial agents with novel modes of action. Shigella pathogenesis is largely intracellular and antibacterial chemicals that preferentially work inside cells may be desirable to limit collateral AMR and block key components of the Shigella infection cycle. Aiming to facilitate the process of identifying antibacterial chemicals that kill intracellular Shigella, we developed a high-throughput screening (HTS) cell-based chemical screening assay. The three-dimensional (3-D) assay, incorporating Shigella invasion into Caco-2 cells on Cytodex 3 beads, was scaled into a 384-well platform for screening chemical compound libraries. Using this assay, we evaluated >500,000 compounds, identifying 12 chemical hits that inhibit Shigella replication inside cells. This simple, efficient and HTS-compatible assays circumvents many of the limitations of traditional screening methods with cell monolayers and may be deployed for antibacterial compound screening for other intracellular pathogens.https://doi.org/10.1038/s44259-025-00110-6 |
| spellingShingle | Voong Vinh Phat Andrew Shih Teong Lim Cristina De Cozar-Gallardo Maria Isabel Castellote Alvaro Demetrio Muñoz Alvarez Elena Fernandez Alvaro Lluis Ballell-Pages Sonia Lozano-Arias Stephen Baker A three-dimensional high throughput assay identifies novel antibacterial molecules with activity against intracellular Shigella npj Antimicrobials and Resistance |
| title | A three-dimensional high throughput assay identifies novel antibacterial molecules with activity against intracellular Shigella |
| title_full | A three-dimensional high throughput assay identifies novel antibacterial molecules with activity against intracellular Shigella |
| title_fullStr | A three-dimensional high throughput assay identifies novel antibacterial molecules with activity against intracellular Shigella |
| title_full_unstemmed | A three-dimensional high throughput assay identifies novel antibacterial molecules with activity against intracellular Shigella |
| title_short | A three-dimensional high throughput assay identifies novel antibacterial molecules with activity against intracellular Shigella |
| title_sort | three dimensional high throughput assay identifies novel antibacterial molecules with activity against intracellular shigella |
| url | https://doi.org/10.1038/s44259-025-00110-6 |
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