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A simple, highly visual in vivo screen for anaplastic lymphoma kinase inhibitors
Frederico S L M Rodrigues1, Xueyan Yang, Masataka Nikaido
1Centre for Regenerative Medicine and Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath BA2 7AY, U.K.
Abstract:
Anaplastic lymphoma kinase (ALK) is an important drug target in many cancers, including lymphoma, neuroblastoma, and lung cancer. Here, we demonstrate proof-of-principle for a novel and inexpensive assay for ALK inhibitor activity and identification in zebrafish. We demonstrate that the human oncogenic ALK fusion, NPM-ALK, drives overproduction of iridophores, a highly visible, shiny pigment cell-type in zebrafish. Treatment with the potent ALK inhibitor, TAE684, fully inhibits production of ALK-dependent iridophores. Using our assay, we test multiple properties of TAE684 in vivo, including efficacy, specificity, and toxicity. We note that TAE684 also inhibits the closely related leukocyte tyrosine kinase (Ltk) that is required for endogenous iridophore development. Similar effects are observed with an independent inhibitor, Crizotinib. Our assay can thus be utilized to identify ALK or LTK inhibitors. Importantly, the natural reflectivity of iridophores lends itself to automation for high throughput assessment of ALK and LTK inhibitor compounds in vivo.
Insights
We developed a novel zebrafish assay to identify anaplastic lymphoma kinase (ALK) inhibitors. This assay uses pigment cells to detect ALK activity and can be automated for high-throughput screening of cancer drugs.
Area of Science:
- Oncology
- Zebrafish models
- Drug discovery
Background:
- Anaplastic lymphoma kinase (ALK) is a key drug target in various cancers.
- Existing methods for ALK inhibitor screening can be costly and complex.
Purpose of the Study:
- To establish a novel, inexpensive, and high-throughput zebrafish assay for detecting anaplastic lymphoma kinase (ALK) inhibitor activity.
- To validate the assay's utility in assessing inhibitor efficacy, specificity, and toxicity in vivo.
Main Methods:
- Utilized zebrafish to model human oncogenic NPM-ALK fusion, which drives iridophore overproduction.
- Administered ALK inhibitors (TAE684, Crizotinib) to assess their impact on iridophore development.
- Evaluated assay automation potential through the reflective properties of iridophores.
Main Results:
- The oncogenic NPM-ALK fusion significantly increased iridophore production in zebrafish.
- Potent ALK inhibitors TAE684 and Crizotinib effectively inhibited ALK-dependent iridophore production.
- The assay demonstrated capability in evaluating inhibitor properties like efficacy, specificity, and toxicity.
Conclusions:
- The developed zebrafish assay serves as a cost-effective platform for identifying and characterizing ALK and leukocyte tyrosine kinase (LTK) inhibitors.
- The assay's potential for automation facilitates high-throughput screening of potential anti-cancer compounds targeting ALK and LTK pathways.
