Orthogonal Drug Pooling Enhances Phenotype-Based Discovery of Ocular Antiangiogenic Drugs in Zebrafish Larvae
Unbiased screening of large randomized chemical libraries in vivo is a powerful tool to find new drugs and targets. However, forward chemical screens in zebrafish can be time consuming and usually >99% of test compounds have no significant effect on the desired phenotype. Here, we sought to f...
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doaj-4d017c23d5524fdcb233e2adce076d522020-11-25T00:34:28ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122019-05-011010.3389/fphar.2019.00508450442Orthogonal Drug Pooling Enhances Phenotype-Based Discovery of Ocular Antiangiogenic Drugs in Zebrafish LarvaeNils Ohnesorge0Temitope Sasore1Daniel Hillary2Yolanda Alvarez3Michelle Carey4Breandán N. Kennedy5UCD School of Biomolecular and Biomedical Sciences, and UCD Conway Institute, University College Dublin, Dublin, IrelandUCD School of Biomolecular and Biomedical Sciences, and UCD Conway Institute, University College Dublin, Dublin, IrelandSchool of Mathematics & Statistics, University College Dublin, Dublin, IrelandUCD School of Biomolecular and Biomedical Sciences, and UCD Conway Institute, University College Dublin, Dublin, IrelandSchool of Mathematics & Statistics, University College Dublin, Dublin, IrelandUCD School of Biomolecular and Biomedical Sciences, and UCD Conway Institute, University College Dublin, Dublin, IrelandUnbiased screening of large randomized chemical libraries in vivo is a powerful tool to find new drugs and targets. However, forward chemical screens in zebrafish can be time consuming and usually >99% of test compounds have no significant effect on the desired phenotype. Here, we sought to find bioactive drugs more efficiently and to comply with the 3R principles of replacement, reduction, and refinement of animals in research. We investigated if pooling of drugs to simultaneously test 8–10 compounds in zebrafish larvae can increase the screening efficiency of an established assay that identifies drugs inhibiting developmental angiogenesis in the eye. In a phenotype-based screen, we tested 1,760 small molecule compounds from the ChemBridge DIVERSet™ chemical library for their ability to inhibit the formation of distinct primary hyaloid vessels in the eye. Applying orthogonal pooling of the chemical library, we treated zebrafish embryos from 3 to 5 days post fertilization with pools of 8 or 10 compounds at 10 μM each. This reduced the number of tests from 1,760 to 396. In 63% of cases, treatment showed sub-threshold effects of <40% reduction of primary hyaloid vessels. From 18 pool hits, we identified eight compounds that reduce hyaloid vessels in the larval zebrafish eye by at least 40%. Compound 4-[4-(1H-benzimidazol-2-yl)phenoxy]aniline ranked as the most promising candidate with reproducible and dose-dependent effects. To our knowledge, this is the first report of a self-deconvoluting matrix strategy applied to drug screening in zebrafish. We conclude that the orthogonal drug pooling strategy is a cost-effective, time-saving, and unbiased approach to discover novel inhibitors of developmental angiogenesis in the eye. Ultimately, this approach may identify new drugs or targets to mitigate disease caused by pathological angiogenesis in the eye, e.g., diabetic retinopathy or age-related macular degeneration, wherein blood vessel growth and leaky vessels lead to vision impairment or clinical blindness.https://www.frontiersin.org/article/10.3389/fphar.2019.00508/fullZebrafishangiogenesiseye vascular systemlibrary screeningdrug poolingorthogonal pooling strategy |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Nils Ohnesorge Temitope Sasore Daniel Hillary Yolanda Alvarez Michelle Carey Breandán N. Kennedy |
spellingShingle |
Nils Ohnesorge Temitope Sasore Daniel Hillary Yolanda Alvarez Michelle Carey Breandán N. Kennedy Orthogonal Drug Pooling Enhances Phenotype-Based Discovery of Ocular Antiangiogenic Drugs in Zebrafish Larvae Frontiers in Pharmacology Zebrafish angiogenesis eye vascular system library screening drug pooling orthogonal pooling strategy |
author_facet |
Nils Ohnesorge Temitope Sasore Daniel Hillary Yolanda Alvarez Michelle Carey Breandán N. Kennedy |
author_sort |
Nils Ohnesorge |
title |
Orthogonal Drug Pooling Enhances Phenotype-Based Discovery of Ocular Antiangiogenic Drugs in Zebrafish Larvae |
title_short |
Orthogonal Drug Pooling Enhances Phenotype-Based Discovery of Ocular Antiangiogenic Drugs in Zebrafish Larvae |
title_full |
Orthogonal Drug Pooling Enhances Phenotype-Based Discovery of Ocular Antiangiogenic Drugs in Zebrafish Larvae |
title_fullStr |
Orthogonal Drug Pooling Enhances Phenotype-Based Discovery of Ocular Antiangiogenic Drugs in Zebrafish Larvae |
title_full_unstemmed |
Orthogonal Drug Pooling Enhances Phenotype-Based Discovery of Ocular Antiangiogenic Drugs in Zebrafish Larvae |
title_sort |
orthogonal drug pooling enhances phenotype-based discovery of ocular antiangiogenic drugs in zebrafish larvae |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Pharmacology |
issn |
1663-9812 |
publishDate |
2019-05-01 |
description |
Unbiased screening of large randomized chemical libraries in vivo is a powerful tool to find new drugs and targets. However, forward chemical screens in zebrafish can be time consuming and usually >99% of test compounds have no significant effect on the desired phenotype. Here, we sought to find bioactive drugs more efficiently and to comply with the 3R principles of replacement, reduction, and refinement of animals in research. We investigated if pooling of drugs to simultaneously test 8–10 compounds in zebrafish larvae can increase the screening efficiency of an established assay that identifies drugs inhibiting developmental angiogenesis in the eye. In a phenotype-based screen, we tested 1,760 small molecule compounds from the ChemBridge DIVERSet™ chemical library for their ability to inhibit the formation of distinct primary hyaloid vessels in the eye. Applying orthogonal pooling of the chemical library, we treated zebrafish embryos from 3 to 5 days post fertilization with pools of 8 or 10 compounds at 10 μM each. This reduced the number of tests from 1,760 to 396. In 63% of cases, treatment showed sub-threshold effects of <40% reduction of primary hyaloid vessels. From 18 pool hits, we identified eight compounds that reduce hyaloid vessels in the larval zebrafish eye by at least 40%. Compound 4-[4-(1H-benzimidazol-2-yl)phenoxy]aniline ranked as the most promising candidate with reproducible and dose-dependent effects. To our knowledge, this is the first report of a self-deconvoluting matrix strategy applied to drug screening in zebrafish. We conclude that the orthogonal drug pooling strategy is a cost-effective, time-saving, and unbiased approach to discover novel inhibitors of developmental angiogenesis in the eye. Ultimately, this approach may identify new drugs or targets to mitigate disease caused by pathological angiogenesis in the eye, e.g., diabetic retinopathy or age-related macular degeneration, wherein blood vessel growth and leaky vessels lead to vision impairment or clinical blindness. |
topic |
Zebrafish angiogenesis eye vascular system library screening drug pooling orthogonal pooling strategy |
url |
https://www.frontiersin.org/article/10.3389/fphar.2019.00508/full |
work_keys_str_mv |
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