PEPITA: Parallelized High-Throughput Quantification of Ototoxicity and Otoprotection in Zebrafish Larvae
Drug-induced hearing injury (ototoxicity) is a common, debilitating side effect of many antibiotic regimens that can be worsened by adverse drug interactions. Such adverse drug interactions are often not detected until after drugs are already on the market because of the difficulty of measuring all...
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Bio-protocol LLC
2024-11-01
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| author | Elizabeth Nilles Ethan Bustad Meng Qin Emma Mudrock Alden Gu Louie Galitan Henry Ou Rafael Hernandez Shuyi Ma |
| author_facet | Elizabeth Nilles Ethan Bustad Meng Qin Emma Mudrock Alden Gu Louie Galitan Henry Ou Rafael Hernandez Shuyi Ma |
| author_sort | Elizabeth Nilles |
| collection | DOAJ |
| description | Drug-induced hearing injury (ototoxicity) is a common, debilitating side effect of many antibiotic regimens that can be worsened by adverse drug interactions. Such adverse drug interactions are often not detected until after drugs are already on the market because of the difficulty of measuring all possible drug combinations. While in vivo mammalian assays to screen for ototoxic damage exist, they are currently time-consuming, costly, and limited in throughput, which limits their utility in assessing drug interaction outcomes. To facilitate more rapid quantification of ototoxicity and assessment of adverse drug interactions that impact ototoxicity, we have developed a high-throughput workflow we call parallelized evaluation of protection and injury for toxicity assessment (PEPITA). PEPITA uses zebrafish larvae to quantify ototoxic damage and protection. Previous work has shown that hair cells (HCs) in the zebrafish lateral line are very similar to human inner ear HCs, meaning zebrafish are a viable model to test drug-induced ototoxicity. In PEPITA, we expose zebrafish larvae to different combinations of drugs, fluorescently label the HCs, and subsequently use microscopy to quantify the brightness of the fluorescently labeled HCs as an assay for ototoxic damage and hair-cell viability. PEPITA is a reproducible, low-cost, technically accessible, and high-throughput assay. These advantages allow many experiments to be conducted in parallel, paving the way for systematic evaluation of drug-induced hearing injury and other multidrug interactions. |
| format | Article |
| id | doaj-art-9f32949b302e463b920dcfa9470a8aaa |
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| issn | 2331-8325 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Bio-protocol LLC |
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| spelling | doaj-art-9f32949b302e463b920dcfa9470a8aaa2025-08-20T02:38:59ZengBio-protocol LLCBio-Protocol2331-83252024-11-01142110.21769/BioProtoc.5105PEPITA: Parallelized High-Throughput Quantification of Ototoxicity and Otoprotection in Zebrafish LarvaeElizabeth Nilles0Ethan Bustad1Meng Qin2Emma Mudrock3Alden Gu4Louie Galitan5Henry Ou6Rafael Hernandez7Shuyi Ma8Center for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USACenter for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USACenter for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USADepartment of Chemical Engineering, University of Washington, Seattle, WA, USACenter for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USACenter for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USACenter for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USADepartment of Pediatrics, University of Washington, Seattle, WA, USADepartment of Otolaryngology-HNS, University of Washington, Seattle, WA, USACenter for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USADepartment of Pediatrics, University of Washington, Seattle, WA, USA, Pathobiology Graduate Program, Department of Global Health, University of Washington, Seattle, WA, USACenter for Global Infectious Disease Research, Seattle Children’s Research Institute, Seattle, WA, USADepartment of Chemical Engineering, University of Washington, Seattle, WA, USA, Department of Pediatrics, University of Washington, Seattle, WA, USA, Pathobiology Graduate Program, Department of Global Health, University of Washington, Seattle, WA, USADrug-induced hearing injury (ototoxicity) is a common, debilitating side effect of many antibiotic regimens that can be worsened by adverse drug interactions. Such adverse drug interactions are often not detected until after drugs are already on the market because of the difficulty of measuring all possible drug combinations. While in vivo mammalian assays to screen for ototoxic damage exist, they are currently time-consuming, costly, and limited in throughput, which limits their utility in assessing drug interaction outcomes. To facilitate more rapid quantification of ototoxicity and assessment of adverse drug interactions that impact ototoxicity, we have developed a high-throughput workflow we call parallelized evaluation of protection and injury for toxicity assessment (PEPITA). PEPITA uses zebrafish larvae to quantify ototoxic damage and protection. Previous work has shown that hair cells (HCs) in the zebrafish lateral line are very similar to human inner ear HCs, meaning zebrafish are a viable model to test drug-induced ototoxicity. In PEPITA, we expose zebrafish larvae to different combinations of drugs, fluorescently label the HCs, and subsequently use microscopy to quantify the brightness of the fluorescently labeled HCs as an assay for ototoxic damage and hair-cell viability. PEPITA is a reproducible, low-cost, technically accessible, and high-throughput assay. These advantages allow many experiments to be conducted in parallel, paving the way for systematic evaluation of drug-induced hearing injury and other multidrug interactions.https://bio-protocol.org/en/bpdetail?id=5105&type=0 |
| spellingShingle | Elizabeth Nilles Ethan Bustad Meng Qin Emma Mudrock Alden Gu Louie Galitan Henry Ou Rafael Hernandez Shuyi Ma PEPITA: Parallelized High-Throughput Quantification of Ototoxicity and Otoprotection in Zebrafish Larvae Bio-Protocol |
| title | PEPITA: Parallelized High-Throughput Quantification of Ototoxicity and Otoprotection in Zebrafish Larvae |
| title_full | PEPITA: Parallelized High-Throughput Quantification of Ototoxicity and Otoprotection in Zebrafish Larvae |
| title_fullStr | PEPITA: Parallelized High-Throughput Quantification of Ototoxicity and Otoprotection in Zebrafish Larvae |
| title_full_unstemmed | PEPITA: Parallelized High-Throughput Quantification of Ototoxicity and Otoprotection in Zebrafish Larvae |
| title_short | PEPITA: Parallelized High-Throughput Quantification of Ototoxicity and Otoprotection in Zebrafish Larvae |
| title_sort | pepita parallelized high throughput quantification of ototoxicity and otoprotection in zebrafish larvae |
| url | https://bio-protocol.org/en/bpdetail?id=5105&type=0 |
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