Hason Martina, Jovicic Jovana, Vonkova Ivana, Bojic Milan, Simon-Vermot Theresa, White Richard M, Bartunek Petr
Laboratory of Cell Differentiation, Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, Czechia.
CZ-OPENSCREEN, Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, Czechia.
Front Pharmacol. 2022 Apr 27;13:893655. doi: 10.3389/fphar.2022.893655. eCollection 2022.
In the last decade, zebrafish have accompanied the mouse as a robust animal model for cancer research. The possibility of screening small-molecule inhibitors in a large number of zebrafish embryos makes this model particularly valuable. However, the dynamic visualization of fluorescently labeled tumor cells needs to be complemented by a more sensitive, easy, and rapid mode for evaluating tumor growth to enable high-throughput screening of clinically relevant drugs. In this study we proposed and validated a pre-clinical screening model for drug discovery by utilizing bioluminescence as our readout for the determination of transplanted cancer cell growth and inhibition in zebrafish embryos. For this purpose, we used NanoLuc luciferase, which ensured rapid cancer cell growth quantification with high sensitivity and low background when compared to conventional fluorescence measurements. This allowed us large-scale evaluation of drug responses of 180 kinase inhibitors in zebrafish. Our bioluminescent screening platform could facilitate identification of new small-molecules for targeted cancer therapy as well as for drug repurposing.
在过去十年中，斑马鱼已成为与小鼠一样强大的癌症研究动物模型。在大量斑马鱼胚胎中筛选小分子抑制剂的可能性使该模型特别有价值。然而，荧光标记肿瘤细胞的动态可视化需要通过一种更灵敏、简便和快速的模式来补充，以评估肿瘤生长，从而实现对临床相关药物的高通量筛选。在本研究中，我们提出并验证了一种临床前药物发现筛选模型，该模型利用生物发光作为检测斑马鱼胚胎中移植癌细胞生长和抑制的读数。为此，我们使用了纳米荧光素酶，与传统荧光测量相比，它能以高灵敏度和低背景确保快速定量癌细胞生长。这使我们能够大规模评估斑马鱼中180种激酶抑制剂的药物反应。我们的生物发光筛选平台有助于识别用于靶向癌症治疗以及药物重新利用的新小分子。
